Aesculus chinensis 

Fructus & semen Aesculus chinensis Regulates the Liver Qi, Expands the middle jiao and harmonizes the Stomach, Contains saponin-aescin which causes a 10 fold increase in adrenocorticotropic hormone (ACTH) and causes a 20 fold increase in plasma corticosterol levels and glucocorticoid.*

Increases DHEA production via pituitary stimulation of adrenals.*

Aesculus chinensis is commonly used in TCM as qi regulating herb and has been used as a stomachic and analgesic in the treatment of distension and pain in chest and abdomen (Zhong Yao Zhi 1984), and the extract of the seeds are used for treating heart diseases (Qian 1996), however modern research shows anti-inflammatory activities of the four main saponins (Wei et al 2004). AH compounds were evaluated for their inhibitory activity against HIV-1 protease (Yang et al 1999).* 

The saponin mixture in Aesculus chinensis, called escin has been used in therapy for peripheral vascular disorders and in the cosmetic field for prevention and treatment of cellulitis because of its anti-inflammatory, anti-edematous and capillary-protective properties (Ogura et al 1975).*

According to patent US 2004/0234633A1, AH inhibits angiogenesis and has significant anti-ageing properties (Kim et al 2004).*

Aesculus hippocastanum L. and Hamamelis virginiana (HV) were found to have strong active-oxygen scavenging activity of and protective activity against cell damage induced by active oxygen. Both AH and HV are proposed as potent plant extracts with potential application as anti-ageing (Masaki et al 1995).*

Cordyceps sinensis

Cordyceps sinensis polysaccharides show evidence that they increase corticosterone production. Other studies suggest they might reduce blood glucose without reducing plasma insulin levels, as well as reduce plasma triglycerides and cholesterol.*

Cordyceps seems to have a beneficial effects on the immune, endocrine, cardiovascular, respiratory, renal, sexual, hepatic, immunologic, and nervous systems.*

Rats fed an extract increased testis weight and sperm count and Cordyceps counteracts the affect of cortisol on the hippocampus.* 

Cordyceps (CS) has a significantly protective effect on spermatogenic damage by cyclophosphamide, the action of cordyceps maybe have relationship with its scavenging free radical and anti-oxidation effects (Jin et al 2008) and the antioxidant activity might mainly derive from reducing intracellular ROS (Gu et al 2008).*

CS can alleviate lipid peroxidation of mice myocardium by increasing active antioxidase and inducing iNOS expression (Li et al 2006).*

Results indicate that after the rats took CS, their physical ability improved remarkably; the activity of Liver SOD appeared superior to the control group; the quantity of MDA turned lower than the control group, which shows that cordyceps sinensis enjoys much stronger influence on the anti-oxidation and anti-free-radical.*

It can also been used to protect the integrality of liver cell membrane; to reduce the escape of enzymes from cells (Zhang 2005).*

The effect of Chinese caterpillar fungus on testis oxidative damage induced by cyclophosphamide in the mice* 

JIN Lei, CHEN Shou-Zhen. Zhong Guo Fu You Bao Jian. 2008; 23(13): 1858-1860.

Objective: To observe the protective effects of cordyceps on the damage of mouse spermatogenesis induced by cyclophosphamide. Results: In cordyceps treatment group, the level of SOD, GSH -Px and sperm density, sperm motility were increased significantly (P<0.05, P<0.01), the level of MDA and sperm abnormality rate were decreased significantly (P<0<05, P<0.01) . Conclusion: Cordyceps has a significantly protective effect on spermatogenic damage by cyclophosphamide, the action of cordyceps maybe have relationship with its scavenging free radical and anti-oxidation effects.*

Cordyceps sp. Reduce Cell Oxidative Damage Induced by Free Radical

GU Yu-xiang, S0NG Yu-wen, FAN Li-qiang, YUAN Qin-sheng. Shi Pin Ke Xue. 2008; 29(2): 387-390.

Objective: Investigating the protective effects of water and ethanol extracts of natural Cordyceps sinensis and Cordyceps militaris and their fermentation preparations against free radical induced oxidative damage in human hepatoma cell line SMMC-7721. Method: The samples were tested through 5 assays: reducing cell growth suppression caused by I-IzOz; inhibition of free radical induced lipid peroxidation; inhibition of intracellular GSH decrease; inhibition of free radical induced DNA oxidative damage and decreasing intracellular ROS levels. Results: Natural and cultured Cordyceps sp. showed different antioxidant ability, and in most cases, their antioxidant activity was dose dependent. Among all 8 extracts, the aqueous fraction of natural C. militaris indicated the highest activity, the inhibition of cell growth decreased 90. 4 %, lipid peroxidation levels decreased 51. 3%, intracellular GSH increased 125 %, DNA oxidation damage decreased 54. 9%, intracellular ROS levels decreased 54. 8% compared with control group at 50 mg/ml. Conclusion: Partial Cordyceps sp. and their fermentation preparations could be regarded as potent natural antioxidants, and the fermented process could affect the antioxidant ability of cultured Cordyceps sp much. The work also presumed that the antioxidant activity of samples might mainly derive from reducing intracellular ROS.*

The antioxidative activities and oxidative injury protective effects of hot-water extracts from cordyceps sinensis mycelia*

LUO Jian-Guang, YANG Xiao-Tong, YANG Qing-Yao, Mi Ke, Feng Huiqin. Xian Dai Yu Fang Yi Xue. 2007; 34(18): 3433-3435.

Objective: To investigate the activities of CS-HWE, the hot-water extracts derived from the liquid-cultured Cordyceps sinensis mycelia, on ant oxidation and oxidative injury protection. Methods: The antioxidant activity was detected by using a Ferric Reducing Antioxidant Power assay(FRAP). The oxidative injury protective effect was assessed by using a PC12 oxidative injury model treated with H2O2, and the survived cells were measured by using MTT assay. Results: The FRAP assay showed the CS-HWE exerted an antioxidative activity, which was(158. 03 5. 39)mmol Fe 2+-1. Its activity was about 75% of that of the whole nature Cordyceps extract was. CS-HWE at concentrations of 50-100 mg/span>-1 could significantly preserve viable PC12 cells after H2O2 treatment, and the protective effect was equivalent to the whole nature Cordyceps extract(P<0.05). Conclusion: The hot-water extract of Cordyceps sinensis fermentative mycelia possess notable antioxidative activities and has the potential to be used as an antioxidative agent.*

Effect of Extract of Cordyceps Sinensis on Lipid Peroxidation of Mice with Viral Myocarditis*

LI Feng, GAO Xing-yu, RAO Bang-fu, LIU Li, CUI Lian-qun. Nan Jin Zhong Yi Yao Da Xue Xue Bao. 2006; 22(1): 21-23. 

OBJECTIVE: To understand the effect and significance of extract of Cordyceps Sinensis on mice with viral myocarditis. METHOD: 100 male BALB/c mice were randomly divided into a control group, an infection group and a group treated with extract of Cordyceps Sinensis. The mice in the infection group and the group treated with extract of Cordyceps Sinensis were inoculated with Cox-sackie B3 virus and were then killed seven and fourteen days respectively after the inoculation. Myocardium SOD, MDA and iNOS expressions were determined and semiquantitative scoring of the pathological changes of the heart was carried out under the optical microscope. RESULT: Compared with the infection group, SOD activity and iNOS expressions in the group treated with extract of Cordyceps Sinensis increased markedly (P<0.01), MDA level decreased (P<0.01) and myocarditis was relieved. CONCLUSION: Extract of Cordyceps Sinensis can alleviate lipid peroxidation of mice myocardium by increasing active antioxidase and inducing iNOS expression.*

The antagonizing lipid peroxide effect of Aweto extract (Cordyceps Sinensis) in acute respiratory distress syndrome*

YAN Dong, LI Lanjuan, DU Weibo, Cao Hongcui. Zhe Jiang Yi Xue. 2006; 28(3): 187-188. 

Objective To study the antagonizing lipid peroxhinaide effect of Aweto water-extract in acute respiratory distress syndrome (ARDS), Methods Twenty-six New Zealand rabbits were randomly divided into three groups: the control group, the ARDS group and the ARDS-treatment group. The ARDS model was established by intravenous injection of oleic acid and in treatment group aweto water-extract was administrated. Superoxide dismutase (SOD) and malondialdehyde (MDA) in plasma were measured and compared in different groups two hours after the model was made. Results The SOD level in model group was lower than that in control group (325. 6818. 20) U/ml vs (370. 1233. 58)U/ml(t=3. 29, P<0.01), The MDA level in model group was higher than that of control group (9. 0832. 46) nmol/ml vs (1, 7080. 98)nmol/ml(t=7. 854, P<0.01). SOD in treatment group (362. 0612. 34)U/ml was significantly higher than that in model group (t=4, 875, P<0.05). MDA in the treatment group (5. 8521. 14)nmol/ml was significantly lower than that in model group (t=3, 535, P<0, 05), Conclusion Aweto have the effect of antagonizing lipid peroxide in ARDS rabbits induced by oleic acid.*

Effect of Cordyceps Sinensis on Free Radical Metabolism and Serum Enzyme in Rats Liver*

ZHANG Min-ling. Xi An Ti Yu Xue Yuan Xue Bao. 2005; 22(6): 70-72. 

Abstract: By setting up the exhaustive swimming rats model, the activity of Liver SOD, the quantity of MDA and the serum GOT are all valued. The results indicate that after the rats took cordyceps sinensis, their physical ability improved remarkably; the activity of Liver SOD appeared superior to the control group; the quantity of MDA turned lower than the control group, which shows that cordyceps sinensis enjoys much stronger influence on the anti-oxidation and anti-free-radical. Besides, their ability of counteracting the free radical of Liver jumped to a high level. Furthermore, cordyceps sinensis can also been used to protect the integrality of Liver cell membrane; to reduce the escape of enzyme from the cell.* 

Prosecutable function of Cordyceps sinensis extracts for hepatic mitochondrial oxidative injuries in diabetic mice*

Zhang Lei, Chen Shun-zhi, Liu Shu-sen. Zhong Guo Lin Chuang Kang Fu. 2006; 10(39): 132-134.

AIM: To analyze the protection of traditional Chinese medicine extract of Cordyceps sinensis (CS) for diabetes-induced oxidative injuries in mice from the angle of mitochondrial respiratory chain. METHODS: This experiment was completed in State Key Laboratory of Biomembrane and Membrane Biotechnology of Institute of Zoology, Chinese Academy of Sciences from March 1998 to July 2000. A total of 32 male Kunming mice were divided into four groups, each containing 8 mice. Normal control group was absent by any treatment, and other mice were treated with alloxan to induce diabetic models. Mice of CS extract group merely took 1 mL CS extract orally, mice of Vitamin E group were offered by 4 mg/kg Vitamin E of subcutaneous injection, and mice of diabetes group were not treated with any protective medicine. We determined the activity of antioxidant enzyme in mitochondria with colorimetry. We also designed to determine the parameter of liver mitochondria about rate of electron transfer to proton pump of mitochondrial respiratory chain complex II + III (H^+/2e). RESULTS: All 32 mice entered the result analysis. (1)The content of glutathione (GSH), activity of GSH peroxidase (GSH-Px), and GSH-Px/ malonaldehyde (MDA) were all decreased obviously in diabetes group than in normal control group, also significantly decreased in diabetes group than in CS extract group and Vitamin E group (P< 0.05-0.01>. (2)MDA content was higher in diabetes group than in normal control group, but lower in CS extract group and vitamin E group than in diabetes group (P< 0.001>. (3) Compared with normal control group, the total and net H^+/2e of mitochondrial respiratory chain complex II + III decreased by 24. 76% and 32. 31% respectively in diabetes group (P< 0.001>. After breeding diabetic mice with extract of CS and Vitamin E respectively, it was found that each data recovered remarkably, which were higher than those of diabetes group (P< 0.001>. CONCLUSION: Free radical generated from mitochondria respiratory chain may be responsible for the liver damage from diabetes. Mitochondrial electron leak, the . decreased coupling of electron transfer and proton pump might be responsible for the decreased mitochondrial efficiency of oxygen utilization. And extract of CS is effective in resisting the oxidative damage on liver mitochondria of diabetic mice.*

Cynomorium songaricum

Cynomorium songaricum elicits significant changes in gonadotrophin levels coupled with a significant increase in ovarian weight and profound folliculogenesis. Numerous primary, secondary, tertiary and antral follicles were present.*

Results show that the average swimming duration of the CS group(II) is longer than that of control and the ratio of blood lactic acid and MDA to swimming duration in group(II) is significantly lower than control (P<0.05).*

The activity of SOD and GSH-Px in skeletal muscle is markedly increased (P<0.05) in group(II). The study indicates that the CS could improve physical endurance and anti-oxidative status in mice (Wang et al 2006).*

CS Group has a strong anti-hypoxia effect and can reduce the damages of cerebral and cardiac muscle tissue induced by altitude low-pressured hypoxia. 

The mechanism may be related to the increase of the SOD activity and inhibition of membrane lipid peroxidation in both cerebral and cardiac muscle tissue, as well as to the decrease of the accumulation of lactic acid in brain tissue and the increase of the protein contents in cardiac muscle tissue (Luo et al 2007).*

CS group has significant improvement on the immuno-pathologic change and free radical metabolism (oxidation-related biomarkers) of ageing rats model induced by D-galactose (Su et al 2009).* 

Administration of CS increased phagocytic activity of macrophages (Chen & Chen P.891).*

The effect of Cynomorium songaricum Rupr on endurance and antioxidative status of skeletal muscle in trained mice*

WANG Zong-bing, GUO Ceng-cheng, FU Yu-ping, LI Dong-zhe. Xi Bei Shi Fan Da Xue Xue Bao: Zi Ran Ke Xue Ban. 2006; 42(4): 100-102.

The effect of Cynomorium songaricum Rupr(CSR) on endurance and antioxidative status is studied in swimming trained mice. 30 Kunming mice are randomly divided into 2 groups, swimming group (I) and swimming + CSR group(II). The results show that the average swimming duration of group(II) is longer than that of group(I)(P<0.01), and the ratio of blood lactic acid and MDA to swimming duration in group(II) is significantly lower than group(I) (P<0.05), the activity of SOD and GSH-Px in skeletal muscle is markedly increased(P<0.05) in group(II). The study indicates that the CSR could improve physical endurance and antioxidative status in mice.*

Pharmacological Effects and Mechanism of Anti-hypoxia Components from Cynomorium songaricum Rupr.*

LUO Junde, ZHANG Ruxue, JIA Zhengping, LI Maoxing, WANG Juan, HU Jing. Zhong Yao Xin Yao Yu Lin Chuang Yao Li. 2007; 18(4): 275-279. 

Objective: To screen the anti -hypoxia components from Cynomorium songaricum Rupr. and to explore its possible anti -hypoxia mechanisms. Methods: The anti -hypoxia components of Cynomorium songaricum Rupr. were extracted and separated by fraction extract and macroporous resin -alcohol gradient elution. The anti -hypoxia components were screened with the closed normobaric hypoxia mice model or under altitude low-pressured hypoxia. The content of water and lactic acid in cerebral tissue, SOD activity, malondialdehyde (MDA) and protein content in both cerebral and cardiac muscle tissue were measured. Results: The survival time of mice under ordinary pressure is significantly longer in the three Cynomorium songaricum Rupr. treated groups than that of blank control group. Furthermore, Part III (from water extract and 95 % alcohol elution through macroporous resin) from the alcohol extract showed the similar effect on increasing the survival time in mice . After administration of Part III, the SOD activity in cerebral and cardiac muscle tissue exposed to hypoxia was increased, the MDA content decreased. Meanwhile, the content of water and lactic acid in cerebral tissue was also decreased while the content of protein in cardiac muscle tissue was increased. Conclusion: Cynomorium songaricum Rupr. can enhance the capacity of hypoxia tolerance. Part III has the strong anti-hypoxia effect and can reduce the damages of cerebral and cardiac muscle tissue induced by altitude low-pressured hypoxia. The mechanism may be related to the increase of the SOD activity and inhibition of membrane lipid peroxidation in both cerebral and cardiac muscle tissue, as well as to the decrease of the accumulation of lactic acid in brain tissue and the increase of the protein contents in cardiac muscle tissue.*

Compare antioxidation of different abstracts of Cynomorium songaricum Rupr. in the aging rats*

LIU Zhao ning, JIA Hai-ying, MA Li-jie, LIU Teng, YANG Li-li, LI Lan-cheng, SIE Jun, SU Xiu –lan. Zhong Guo Ming Zu Yi Yao Za Zhi. 2009; 15(2): 42-43. 

Objective: To Compare antioxidation of different abstracts Cynomorium songaricum Rupr. in the D-gal-induced aging rots. Methods: D-gal-induced aging rats are fed on different abstracts of Cynomorium songaricum Rupr. Daily. After 8 weeks, the activities of CAT, total antioxidation, and GSH-PX in blood were determined. Results: Compared with aging group, the activities of CAT, total antioxidation, and GSH-PX in rats fed on abstracts of Cynomorium songaricum Rupr. Increased evidently, especially the ethanol abstract increased significantly. Conclusion: the ethanol abstract of Cynomorium songaricum Rupr. has significantly abilities of antioxidation and retarding aging process.*

Effect of Cynomorium Songaricum Rupr. chewable tablets on immunopathologic changes and free radical of spleen in aging rats*

SU Yun, LIU Yong-Qi, WU Jian-Jun, Zhang Yi, Yan Chunlu, Nie Lei. Zhong Guo Lao Nian Xue Za Zhi. 2009; 29(8): 927-929. 

Objective: To explore the anti-aging effect and mechanism of the Cynomorium water extract chewable tablets and Cynomorium polysaccharide chewable tablets. Methods: The Cynomorium polysaccharide and Cynomorium water extract were extracted and prepared chewable tablets. Wistar rats were randomly divided into normal control, aging model, Vit E, Cynomorium polysaccharide chewable tablets and Cynomorium water extract chewable tablets groups. D-galactose (D-gal) was injected to establish the aging model. After drug intervention, the proliferation activity of rat spleen lymphocytes stimulated by ConA was detected by MTF assay and the spleen index was measured. The pathological morphological changes of spleen were observed. The activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA) and nitric oxide (NO) in serum of rats were detected. Results: Compared with the normal control group, the spleen index, the proliferation activity of rat spleen lymphocytes stimulated by ConA of aging model rats were significantly lower, the spleen had obvious pathological injury, and the activity of SOD was significantly lower, the content of MDA and NO increased significantly (allP<005). Cynomorium chewable tablets made above abnormal indexes significant improve (P< 005). Conclusions: The extraction of Cynomorium chewable tablets has significant improvement on the immunopathologic change and free radical metabolism of aging rats model induced by D-gal.*

Ganoderma lucidum

Ganoderma lucidum in TCM is said to nourish the Heart and calm the Shen. as well as tonifying Qi. It is mostly known as an immune and a DNA polymerase activity enhancer, also called Reishi, the extract contains high levels of adenosine ATP and increases cAMP concentration in cardiac muscles.*

Middle and high doses of GL can reduce the free radical and accelerate the clearance of these free radicals. It has certain anti-ageing effects.*

The contents of SOD in GL group were significantly higher than those in control group and the low doses GS group (P<0.05), and the contents of MDA were significantly lower than those in control group (Jing et al 2008).*

GL could prevent tBOOH-induced cell oxidative injury in vivo and in vitro. GL increased the survival rate of macrophages injured by tBOOH.*

GL could protect the cell organelles such as mitochondria and endoplasmic reticulums against injury. The mitochondria, which were the major site producing ROS and also subjected to great injury by ROS, were significantly protected (You & Lin 2002). GL improves learning and memory ability (Wang et al 2004).*

Effect of high germanium ganoderma on the activity of serum superoxide dismutase and malondialdehyde of natural senescent rats* 

JING Chang-qin, ZHANG Xiu-hua, LIU Rui, Huo Xiao Dong. Xin Xiang Yi Xue Yuan Xue Bao. 2008; 25(2): 124-126.

Objective To investigate the effect of high germanium ganoderma on the activity of serum superoxide dismutase(SOD) and malondialdehyde(MDA) of natural senescent rats. Methods Forty natural senescent rats were randomly divided into four groups : control group, low, middle and high doses of high germanium ganoderma group. High germanium ganoderma power was diluted to the concentration of 20 g ጞ-1, 40 g ጞ-1 and 80 gጞ-1, then the low, middle and high doses of high germanium ganoderma group were given the different concentration solution with intragastric administration respectively, and the control group was given distilled water. The solution volume of all rats with intragastric administration was 10 mL ᫧^-1, once a day, then killed by breaking head, and collected the blood to determine the content of SOD and MAD. Results The contents of SOD in middle and high doses of high germanium ganoderma group were significantly higher than those in control group and low doses of high germanium ganoderma group (P<0.05), and the contents of MDA were significantly lower than those in control group and low doses of high germanium ganoderma group(P< 0.05). There is no significant difference of those between middle doses of high germanium ganoderma group and high doses of high germanium ganoderma group and between control group and low doses of high germanium ganoderma group (P<005). Conclusion The dose of high germanium ganoderma should be taken moderately. Middle and high doses of high germanium ganoderma can reduce the free radical and accelerate the clearance of the free radical. It has certain anti-aging effect.*

Epimedium brevicornum

Tonifying kidney with Epicedium on mRNA expression of estrogen receptor α and β of the amygdala and the cortical parietal lobe in male rats.*

The expression of ERβ in group EH was higher than that of control (P<0.01), the expression ERβ of amygdala in group B was higher than the control group (P<0.05).*

Tonifying Kidney may perhaps improve the cognitive function through regulating ERβ gene. The result disagrees with the traditional theory of Chinese medicine (Cai et al 2006).*

The inhibition of ERα decreased cell growth and respiratory chain capacity. The inhibition of ERα produced a greater effect in the oxidative cell model, decreasing the mitochondrial mass and the phosphorylating respiration (Mirebeau-Prunier et al 2010).* 

Icariin

From the age of 30 yrs. on, the (free) testosterone [(F)T] levels decrease continuously with age, mean total T level at age 70 yrs., being only 2/3rds of the mean level at age 25 yrs., whereas mean FT level is only 40% of the mean level in young adults. However, inter-individual variations are wide and whereas at least 30% of men over age 70 yrs. have clearly subnormal (F)T levels. androgen deficiency in young adults, such as decrease in muscle mass and strength, increase in abdominal, mainly visceral fat with insulin resistance and atherogenic lipid profile, decrease in libido and sexual hair, osteopenia, decrease in cognitive performances, insomnia, excessive sweating and decrease in general well-being.*

Partial Androgen Deficiency in Aging Males (PADAM) is characterized by an age-related decrease in testicular secretion of testosterone. PADAM contributes to the age-related decrease in bone mineral density (BMD) and the prevalence of hypogonadism increased with ageing.* 

Hypogonadal men were older and heavier (due to a higher fat body mass) and had lower concentrations of 17-oestradiol and androstenedione than men with normal androgenic activity. They demonstrated lower muscle strength, impaired static and dynamic balance, a higher risk of falls, and lower lean muscle mass (Szulc et al 2003).*

lcariin possesses the anti-partial androgen deficiency effects, and is beneficial to partial androgen deficiency in aging males (PADAM).*

Icariin improved the condition of reproductive organs and increased the circulating levels of testosterone. Icariin treatment also improved the steady-state serum BGP and might have promoted bone formation. At the same time, it decreased the serum levels of StrACP and might have reduced the bone resorption. Lastly, icarrin suppressed the extent of apoptosis of penile cavernosal smooth muscle cells (Zhang & Yang 2006).*

Genes expressions of pro-inflammatory cytokines and receptors are upregulated in the hippocampus of aged rat, which may eventually contribute to disequilibrium of the network of pro-and anti-inflammatory cytokines and receptors and disequilibrium of response of pro-and anti-inflammatory system. Icariin may interfere with Լspan class=SpellE>inflamm-ageingՠvia rebuilding a novel equilibrium of the network of pro-and anti-inflammatory cytokines and receptors (Xia et al 2008). Icariin can improve the learning and memory abilities in Ab25-35-induced Alzheimerճ disease (AD) rats.*

The mechanisms may be related to its improving effect on the cholinergic transmitter metabolism of central nervous system, resulted from increment of acetylcholinesterase (AChE) and choline acetyhransferase (ChAT) expressions, anti-oxidative capability and inhibitory activity of NOS.*

The apoptosis rate of rat lymphocytes gradually has been increased and correlated with the time of ageing. NF-κB plays a key role in ageing. Icariin could inhibit the excessive apoptosis of lymphocytes in aged rats (Liu et al 2007).*

Icariin can suppress the excessive apoptosis of splenic lymphocyte in aged rats and activate Rel/NF-κB/ I B/IKK and their signal transduction pathway to up-regulate NF-κB through adjusting I Bε and I Ba, which is the mechanism of Icariin in rebuilding the immune homeostasis of T lymphocyte apoptosis and retarding immuno-senescence Liu et al 2006).*

Study on Extraction and Antioxidation Effects of Flavonoids from E. wushanense*

XIE Juan-ping. An Hui Nong Ye Ke Xue. 2008; 36(35): 15532-15534. 

Objective: The purpose was to study the extraction of effective components from E. wushanense and oxidation resistance of flavonoids (diphylloside A, epimedoside A, epimedinC, ieariin, epimedoside C, iearisoside A and desmethylanhydroiearitin). Method: The column chromatographic separation was used to get various flavonoids. With in vitro antioxidation system as model and DPPH free radical as scavenging object, the antioxidant activity of flavonoids was determined by spectrophotometry with Vc as CK. Result: The results indicated that the extracting solution from E. wushanense and seven kinds of flavonoids had scavenging effect on DPPH free radical and strong antioxidant capacity, the antioxidant capacity of Desmethylanhydroicaritin was the strongest than other, secondly epimedoside C, which scavenging effect on DPPH free radical had close related to structure of flavonoids. Scavenging effects of the two flavonoids were high as well as Vc. Conclusion: This research will provide the scientific basis for application of E. wushaneme in cosmetic, health products and medicine.*

Inflamm-aging related genes expressions in aged rat hippocampus and Icariin intervention outcome *

XIA Shi-jin, SHEN Zi-yin DONG Jing-cheng, LIU Xiao-yu, HUANG Jian-hua, Wang Hai-dong, WU Bin, CHEN Wei-hua. Lao Nian Yi Xue Yu Bao Jian. 2008; 14(6): 340-344.

Objective: This work was to study mechanisms of the inflammatory cytokine network in the development of inflamm-aging and the effect and mechanism of Icariin (Ica) on it. Methods: Healthy male Sprague Dawley rats were divided into 3 groups (10 rats each group): 4m group with 4 months rats, 24m group with 24 months rats with distilled water intragastric administration and 24m+Ica group with 24 months rats treated with Ica by intragastric administration for 3 months from 21 months. Gene expression profiles of rat hippocampal tissues from each group were analyzed by using inflammatory cytokines and receptors cDNA microarrays. Differentially expressed genes were screened. At the same time, inflamm-aging-related inflammatory cytokine protein expression levels of TNF-a, IL-6 and IL-10 in rat hippocampal tissues were evaluated by Enzyme Linked Immunosorbent Assay. Results Compared with 4m group, there were 9 differentially up-regulated expressed pro-inflammatory genes and 6 down-regulated expressed anti-inflammatory genes in rat hippocampal tissues of 24m group. Compared with 24m group, there were 6 up-regulated expressed anti-inflammatory genes and 9 down-regulated expressed pro-inflammatory genes in 24m+Ica group. Compared with 4m group, protein levels of TNF-a and IL-6 in the supernatant from rat hippocampal tissues of 24m group increased significantly (P<0.01). Compared with 24m group, protein levels of TNF-a and IL-6 in the supernatant from rat hippocampal tissues of 24m+Ica group decreased significantly (P<0.01), while protein level of IL-10 in 24m+Ica group was significantly higher than that in 24m group (P< 0.01). The ratio of TNF-a to IL-10 and IL-6 to IL-10 in 24m group increased significantly compared with that in 4m group (P<0.01), and decreased significantly in 24m+Ica group compared with that in 24m group (P<0.01). Conclusions Genes expressions of pro-inflammatory cytokines and receptors are upregulated in the hippocampus of aged rat, which may eventually contribute to disequilibrium of the network of pro-and anti-inflammatory cytokines and receptors and disequilibrium of response of pro-and anti-inflammatory system and high inflammatory response state, thus providing novel mechanistic insight into inflamm-aging. Ica may interfere with inflamm-aging via rebuilding a novel equilibrium of the network of pro-and anti-inflammatory cytokines and receptors and equilibrium of response of pro-and anti-inflammatory system, which was caused by down-regulated expressions of pro-inflammatory cytokine and receptor genes, and upregulated expressions of anti-inflammatory cytokine and receptor genes.*

Protective effects of icariin on learning and memory dysfunction induced by amyloid b-protein fragment 25 –35 *

NIE Jing, LUO Yong, HUANG Xie-Nan, LU Yuan-Fu, SUN An-Sheng, GONG Qi-Hai, SHI Jing-Shan. Zhong Guo Yao Li Xue Yu Du Li Xue Za Zhi. 2008; 22(1): 31-37.

AIM: To investigate the protective effects of icariin on the learning and memory abilities in Alzheimerճ disease (AD) model rats, and explore its potential mechanisms. METHODS: AD model was induced by hippocampus-injection with amyloid b-protein fragment 25-35 (Ab25 -35) 10 mg. From the next day, icariin 30, 60 or 120mg᫧^-1 was administered by gavage for 14 d. The abilities of spatial learning and memory were tested by Morris water maze from d 15 to d 19. The expressions of acetylcholinesterase (AChE) and choline acetyhransferase (CHAT) in hippocampus were detected by immunohistochemistry and quantitated analysis by BI2000 image analysis system. The activities of nitric oxide synthase (NOS), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in hippocampus were assayed by photochemical method in d 20. RESULTS: Compared with AD model rats, icariin treatments significantly caused the decreases in escape latency and searching distance in the Morris water maze test, increased the expressions of AChE and CHAT, enhanced the activities of SOD and GSH-PX, and decreased the activity of NOS in hippocampus. CONCLUSION: Icariin can improve the learning and memory abilities in Ab25-35-induced AD rats. The mechanisms may be related to its improving effect on the cholinergic transmitter metabolism of central nervous system, resulted from increment of AChE and ChAT expressions, anti-oxidative capability and inhibitory activity of NOS.*

Antioxidative protective effect of icariin on the FeSO4/H 2O 2-damaged human sperm based on confocal raman micro-spectroscopy.*

Huang ZS, Xiao HJ, Qi T, Hu ZM, Li H, Chen DL, Xu YL, Chen J. J Huazhong Univ Sci Technolog Med Sci. 2014 Oct;34(5):755-60. doi: 10.1007/s11596-014-1348-3. Epub 2014 Oct 16.

Oxidative stress is implicated in male infertility and significantly higher reactive oxygen species are detected in 25% of infertile males. Although different agents of various alternative medicines, including traditional Chinese medicine, have been tried with varying success, evidence remains limited on whether and how much herbs or supplements might help increase the anti-oxidant ability of the sperm.* 

This study examined the anti-oxidative effects of icariin, a flavonoid isolated from Herba Epimedii, on the human sperm. We prepared the FeSO4/H2O2-damaged human sperms, which were co-cultured with icariin in vitro, and then observed the changes of the sperm by employing Raman micro-spectroscopy. The results showed that Raman mapping with a 514 nm excitation laser allowed clear differentiation of the nucleus, neck, and, in particular, the mitochondria-rich middle piece of a human sperm cell.* 

The effect of icariin on different organelles of the sperm was quantified by localized spectral Raman signatures obtained within milliseconds, and icariin could keep the "Raman fingerprint" of the human sperm the same as the control groups, suggesting that icariin could protect the human sperm from being damaged by FeSO4/H2O2. Icariin may serve as a tonifying and replenishing agent of herbal origin for enhancing reproductive functions.*

In vitro Antioxidant Properties of Epicedium flavonoids*

YANG Jian-xiong, YANG Li-xia, YANG Chen, BAO Yu, HUANG Ya-ya. Tian Ran Chan Wu Yan Jiu Yu Kai Fa. 2007; 19(B05): 72-75. 

To study the in vitro antioxidant activity of the Epicedium flavonoids (EF), the scavenging effect of EF on hydroxyl radicals(᠏H) was studied by salicylic acid method, and serf-oxidation inhibition was studied using the pyrogallol. The contents of MDA in 4 organs, and in mitochondria and microsome of liver were measured by TBA assay. The extent of erythrocatalysis and the swelling extent of mitochondria of liver were detected by spectrophotometric methods. The result showed that EF had the ᠏H and O2 ፊscavenging activity. It could inhibit the generation of MDA, the swelling of mitochondria, and the hemolysis of red blood cells. The conclusion was that EF have the antioxidant activity.*

Comparative studies of the apoptosis rates of lymphocytes in the progress of aging and interventional effect of EF on aged rats*

LIU Xiao-Yu, SHEN Zi-Yin, WU Bin, HUANG Jian-Hua, XIA Shi-Jin, CHEN Wei-Hua. Zhong Guo Mian Yi Xue Za Zhi. 2007; 23(9): 803-805.

Objective: Compare the apoptosis rates of lymphocytes in the progress of aging rats and interventional effect of EF. Methods: The lymphocytes derived from SD rat spleen were employed as the studied cells and were observed by flow cytometry. The differences of apoptosis rates in groups from 3d to 27m were emphasized in the study. The aim of this study of effect of EF on excessive apoptosis of aging rats was to found foundation for the following study. Results:3-day newborn rat group had the lowest apoptotic percentage. The extent of apoptosis correlated with prolongation of age but an exception in 10m group, which had a lower apoptotic percentage than 4m group. The highest apoptotic percentage existed in 27m. When intervened with EF, the apoptosis ratio of 27m group apparently decreased to the level between 10m and 18m. All of the data were statistically significant(P<005 orP<0001). Conclusion: The apoptosis rate of rat lymphocytes gradually has been increased and correlated with the time of aging. NF-κB plays a key role in aging. EF could inhibit the excessive apoptosis of lymphocytes in aged rats.*

Mechanism of Epimedium Flavonoids in Regulating Immuno-senescence via Nuclear Factor-kappa B Related Signal Transduction Pathway* 

LIU Xiao-yu, SHEN Zi-yin, HUANG Jian-hua, Chen Weihua. Zhong Guo Zhong Xi Yi Jie He Za Zhi. 2006; 26(7): 620-624. 

Objective: To explore the mechanism of Epimedium flavonoids (EF) in regulating immunosenescence via nuclear factor-kappa B (NF-κB) related signal transduction pathway. Methods (1) The apoptosis index (AI) of splenic lymphocyte in aged rats was monitored by flow cytometry that of young rats was taken as control. (2) The differential expression profile of NF-κB related signals in aged rats allocated in the control group (aged rats, group A), the EF treated group (group B), the PDTC (a NF-κB inhibitor) treated group (group C) and the PDTC plus EF treated group (group D), was determined and the main significant molecules in them were analyzed with gene microarray of 96 genes related to NF-κB signal pathway. Results: Excessive apoptosis of T lymphocyte cell was seen in aged rats, and it was significantly suppressed in group B and D. In group B, 73 genes were up-regulated to different extent, including 10 of the NF-κB/Rel/IB gene family, transduction signal molecule member of N IK/IKK/I B/Rel/NF-κB, NF-κB regulatory target genes, trans-membrane receptors, transcription factors, and receptor protein, etc. But the up-regulation on NF-κB gene family could not be seen in group C and that on others were also alleviated, while in the group D, the NF-κB gene family and its related transduction pathway were still activated to some extent. The NF-κB gene family showed a markedly common feature after EF intervention, either used alone or in combination with PDTC, i. e. the significant upregulated NF-κB1, NF-κB2, Rel B and I Bε, and activated NIK/IKK/I B/Rel/NF-κB pathway. Conclusion EF can suppress the excessive apoptosis of splenic lymphocyte in aged rats and activate Rel/NF-κB/ I B/IKK and their signal transduction pathway to up-regulate NF-κB through adjusting I Bε and I Ba, which may be the essential mechanism of EF in rebuilding the immune homeostasis of T lymphocyte apoptosis and retarding immunosenescence.*

Cistanche deserticola with 10% standardized β-Sitosterol

The Cistanches extract (CE) showed strong antioxidant activity in photo-chemiluminescence (PCL) experiment (Li et al 2009).*

In CE group MDA content in heart, liver and brain was significantly decreased. In CE (40 mg᫧^-1) group telomerase activity in heart and brain was significantly increased, lymphocyte proliferation, phagocytosis of peritoneal macrophages and peripheral blood IL-2 content obviously enhanced. CE may delay ageing, which may be related to antagonizing free radical injury and enhancing immunity of ageing mice (Zhang H-Q et al 2008).*

The liver structure of the control group was injured in swim test, the isoenzyme of LDH4 and LDH5 were high, the liver glycogen was poor, NOS3 was decreased compared with the normal control group (P<0.05).*

The liver structure of the CE experimental group was good, LDH5 isoenzyme was low, the glycogen was rich and expression of NOS3 was upregulated compared with the sport control group (P<0.05) (Yang H-x et al 2008).*

Study on the Antioxidant Activities of Phenylethanoid Glycosides from Cistanche deserticola*

LI Li, Shi Dongfang, Gui Yuge, Liu Chunming. An Hui Nong Ye Ke Xue. 2009; 37(32): 15835-15836.

Objective: The research aimed to study systematically on the antioxidant activities of phenylethanoid glycosides from Cistanche deserticola. Method: Two model systems, the free radical 2, 2-diphenyl-l-pierylhydrazyl (DPPH᩠assay and the photochemilumineseenee (PCL) assay, were used to measure the antioxidant activity of the C. deserticola extract, acteoside, isoacteoside and eehinaeoside. Result: The results showed that the total phenolic contents (TPCs) in echinacoside, acteoside, isoacteoside and 80% ethanol extract of C. deserticola were 753. 95, 659. 94, 356. 14 and 14. 73 mg gallic acid/g sample. The C. deserticola extract showed strong antioxidant activity in PCL experiment, followed by echinacoside, aeteoside and isoacteoside. However, eehinacoside exhibited strong antioxidant activity in DPPH experiment, followed by acteoside, isoacteoside and the C. deserticola extract. Conclusion: This research could provide references for the pharmacology and biological activity study of pheny]ethanoid glycosides from C. deserticola.*

Effect of Cistanche tubulosa (Scheuk) Whight acteoside on telomerase activity and immunity of aging mice*

ZHANG Hong-Quan, WENG Xiao-Jing, CHEN Li-Li, LI Xin. Zhong Guo Yao Li Xue Yu Du Li Xue Za Zhi. 2008; 22(4): 270-273.

AIM: To study the effect of Cistanche tubulosa (Scheuk) Whight acteoside (CTWA) on malondialdehyde (MDA) content, telomerase activity in heart, liver and brain tissues and immune function of experimentally aging model mice. METHODS: Mice were given sc 10% D-galactose 10 mg᫧^-1, once daily for 8 weeks to establish model of aging mice. CTWA 10, 20 and 40 mg᫧^-1 were given ig, respectively, from the ninth week once daily for 2 weeks. The MDA content and telomerase activity were measured with thiobarbituric acid colorimetric method and PCR-ELISA, respectively. The lymphocyte proliferation was measured by [^3H] TdR incorporation test. Phagocytosis of peritoneal macrophages was tested by neutral red test. The interleukin-2 (IL-2) content in peripheral blood was measured by radioimmunoassay. RESULTS: In model group, MDA content was significantly increased in heart, liver and brain, telomerase activity was significantly decreased in heart and liver, and lymphocyte proliferation, phagocytosis of peritoneal macrophages and blood IL-2 content were obviously decreased. After treatment with CTWA for 2 weeks, MDA content in heart, liver and brain was significantly decreased. In CTWA 40 mg᫧^-1 group telomerase activity in heart and brain was significantly increased, lymphocyte proliferation, phagocytosis of peritoneal macrophages and peripheral blood IL-2 content obviously enhanced. CONCLUSION: CTWA may delay aging, which may be related to antagonizing free radical injury and enhancing immunity of aging mice.*

Experimental Study of Anti-Sports Fatigue Effect Mechanisms of Cistanche Deserticola*

YANG Hong-xin, YANG Yong, YAN Xiao-hong. Zhong Guo Zhong Yi Yao Xin Xi Za Zhi. 2008; 15(4): 24-25.

Objective To study the effect of cistanche deserticola on lactate dehydrogenase (LDH) isoenzyme, glycogen and nitric oxide synthase 3 (NOS3) of liver in the mice burden swimming and explore relevant molecular mechanisms of anti-sports fatigue. Methods The mice were divided into the normal control group, the sport control group and the cistanche deserticola experimental group. Each mice of the normal control group and the sport control group was given saline 0. 2 mL per day. Each mice of the cistanche deserticola experimental group was given water decoction of cistanche deserticola 0. 2 mL (3 g/kg) per day. The administrations were for 15 days. The burden swimming for 90 minutes was carried out on the mice of the sport control group and the cistanche deserticola experimental group at 1 hour after the last administration. Livers of the mice were removed after 10 hours of swimming. One part of liver was fixed in the neutral formalin liquid to prepare the paraffin sections and the others was used for measurement of LDH activity. The structure of the liver was observed by staining of HE and the liver glycogen were measured by staining of glycogen. NOS3 was examined by S-P immunohistochemical method. Results The liver structure of the sport control group was injured seriously, the isoenzyme of LDH4 and LDH5 were higher, the liver glycogen were poor, NOS3 was decreased compared with the normal control group (P<0.05). The liver structure of the cistanche deserticola experimental group was good, LDH5 isoenzyme was low, the glycogen was rich and expression of NOS3 was upregulated compared with the sport control group (P<0.05). Conclusions Cistanche deserticola could decrease LDH5, protect the liver of the mice burden swimming and accelerate glycogen accumulation by increasing expression of NOS3 to protect liver and improving physical capacity recovery.*

Antioxidant property of general cistanosides in human HL-60 cell line*

Mourboul ABLISE, MAO Xin-min, Rena KASIMU, MA Shu yan, Leininger Muller B2, Grard SIEST. Zhong Guo Yao Li Xue Tong Bao. 2008; 24(3): 362-364. 

Aim To investigate the antioxidant property of general cistanosides (GCs)in human HL-60 cell line. Methods GCs was obtained from the acetonic extract of Xinjiang Cistanch salsa; its in vitro antioxidant activity was studied in human HL-60 cell line oxidation system. The generation of cellular free radical (ROS) was followed by oxidation of DCFH-DA probing into its oxidized product DCF. The Ginkgo biloba extract EGB 761 was used as positive control. Results At the concentration of 0. 25, 0. 50 g ᠌^-1 and 0. 75 g ᠌^-1, GCs inhibited the ROS formation by 56% and 62% and 67%, respectively. The ROS inhibitory activity of GCs was similar to that of EGB 761. Conclusion GCs has relatively high antioxidant activities in human HL-60 cell line.*

Cnidium monnieri

Osthol and total coumarins of Fructus Cnidii were given to the model of Kidney-Yang Deficiency rats induced by hydrocortisone acetate injection, the changes of serum concentration of thyroxine T3, reverse T3 (rT3), T4 and thyroid stimulating hormone (TSH) were observed. Results showed that in comparison with normal rats, all of the criteria mentioned above were lowered significantly (P < 0.01) in model rats. After treatment, the criteria observed were all increased significantly (P < 0.01 or P < 0.05). It was suggested that the osthol and total coumarins of Fructus Cnidii could elevate the pituitary-thyroid axis function of Kidney-Yang Deficiency rats. Studies show that She Chuang Zi can stimulate the secretion of 17 beta-estradiol and stimulate the ovulatory process by stimulating the production of CINC/gro, IL-1 beta and TNF alpha in vitro. As a treatment for ovulatory disorders, She Chuang Zi may have stimulatory effects on both steroidogenesis and the ovulatory process.*

Subcutaneous injections in rats increased the weight of the uterus, ovaries and testes. *

Morindae officinalis

Morindae (MO) increased lean muscle body weight and an improvement in physical performance measured in mice (Chen & Chen 2004 P.892).*

MO increased plasma levels of corticosteroids by the stimulation of the pituitary and adrenal cortex (Guo Wai Yiɱ990). A decoction of Semen Cuscutae, Rhizoma Curculiginis and Radix Morindae Officinalis is an effective promoting preparation for sperm motility and the membrane function stabilized. Therefore, it may be beneficial in treating male sterility and may raise success rate in artificial insemination (Peng et al 1997).*

The extract from M. officinalis can significantly intervene in lipid peroxidation in sperm membrane by guarding against oxidation, and protect the structure and function of sperm membrane, it is one of the mechanisms for treating male infertility and asthenozoospermia with MO (Yang et al 2006).*

MO has the beneficial effect on the concentration, morphology and motility of sperm. The concentration of total sperm, the motility and normality of spermatozoa was significantly different when compared with the control group. (Choi et al 2005).*

Icariin attenuates LPS-induced acute inflammatory responses: involvement of PI3K/Akt and NF-kappaB signaling pathway.*

Xu, C-Q., Liu, B-J., Wu, J-F., Xu, Y-C., Duan, X-H., Cao, Y-X. & Dong, J-C. European journal of pharmacology. Vol 642, 2010 Sep. Pp. 146-53. PMID: 20519138     

The mechanism underlying the attenuation of LPS-induced lung inflammation  by icariin in vivo and in vitro were investigated. The anti-inflammatory effects of icariin on LPS-induced acute inflammatory and the molecular mechanism were demonstrated.*

Pretreatment with icarrin (20mg/kg) could attenuate acute lung inflammation  by inhibiting mRNA expressions of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), metalloproteinase cycloxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) in the lung of LPS-treated mice. In addition, icariin suppressed the secretion of TNF-alpha, prostaglandin E2 (PGE(2)) and nitric oxide (NO) as well as NF-kappaB p65 activation. Furthermore, decreased myeloperoxidase (MPO) activity was observed in the lung tissue and LPS-induced cytotoxicity in the RAW 264.7 macrophages cells was also markedly attenuated by icariin. Western blotting analysis and confocal microscopy showed that icariin pretreatment reduced the nucleus transportation and constant level of NF-kappaB p65 in the RAW 264.7 macrophage cells.*

However, the protective effects of icariin were reversed by a PI3K/Akt inhibitor (wortmannin). In vitro and in vivo results suggested that activation of the PI3K/Akt pathway and the inhibition of NF-kappaB were involved in the protective effects of icariin on LPS-induced acute inflammatory responses.*

*These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.