In vitro biological assessment of berberis vulgaris and its active constituent, berberine: antioxidants, anti-acetylcholinesterase, anti-diabetic and anticancer effects.*

Abeer E Abd El-Wahab, Doaa A Ghareeb, et al. BMC Complementary and Alternative Medicine 2013, 13:218 doi:10.1186/1472-6882-13-218

Berberis vulgaris is a well known plant with traditional herbal medical history. The aims of this study was to bioscreen and compare the in vitro biological activity (antioxidant, cholinergic, antidaibetic and the anticancer) of barberry crude extract and berberine active compound.*

Methods: The effect of B. vulgaris extract and berberine chloride on cellular thiobarbituric acid reactive species (TBARS) formation, diphenyle--alpha-picrylhydrazyl (DPPH) oxidation, cellular nitric oxide (NO) radical scavenging capability, superoxide dismutase (SOD), glutathione peroxidase (GPx), acetylcholinesterase (AChE) and alpha-gulcosidase activities were spectrophotometrically determined. On the other hand, the effect of extract and berberine as anticancer was estimated on three different cell lines which were MCF-7, HepG-2, and Caco-2 cells by using neutral red uptake assay which compared with control normal cells (PBMC).*

Results: Our results showed that barberry crude extract contains 0.6 mg berberine/mg crude extract. Barberry extract showed potent antioxidative capacity through decreasing TBARS, NO and the oxidation of DPPH that associated with GPx and SOD hyperactivation. Inhibitory effect of berberis crude extract on alpha-glucosidase was more potent than that of berberine chloride, while both had the same AChE inhibitory effect. Besides, different concentrations of both berberine chloride and barberry ethanolic extract showed to have no growth inhibitory effect on normal blood cells (PBMC). Otherwise, both berberine chloride and barberry ethanolic extract showed to have inhibitory effect on the growth of breast, liver and colon cancer cell lines (MCF7, HepG2 and CACO-2, respectively) at different incubation times starting from 24 hrs up to 72 hrs and the inhibitory effect increased with time in a dose dependant manner.*

Conclusion: This work demonstrates the potential of the barberry crude extract and its active alkaloid, berberine, on suppressing lipid peroxidation, suggesting a promising use in the treatment of hepatic oxidative stress, Alzheimer and idiopathic male factor infertility. Beside, berberis vulgaris ethanolic extract is safe non-toxic extract as it was not inhibit the growth of PBMC that can induce cancer cell death that could return to its powerful antioxidant activity.*

The alkaloid Berberine inhibits the growth of Anoikis-resistant MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell cycle arrest.*

Kim JB, Yu JH, Ko E, Lee KW, et al. Phytomedicine. 2010 May;17(6):436-40. doi: 10.1016/j.phymed.2009.08.012. Epub 2009 Oct 2.

Berberine is a pure phenanthren alkaloid isolated from the roots and bark of herbal plants such as Berberis, Hydrastis canadensis and Coptis chinensis. Berberine has been established to inhibit the growth of breast cancer cells, but its effects on the drug resistance and anoikis-resistance of breast cancer cells have yet to be elucidated. Anoikis, or detachment-induced apoptosis, may prevent cancer progression and metastasis by blocking signals necessary for survival of localized cancer cells.*

Resistance to anoikis is regarded as a prerequisite for metastasis; however, little is known about the role of berberine in anoikis-resistance. We established anoikis-resistant cells from the breast cancer cell lines MCF-7 and MDA-MB-231 by culturing them on a Poly-Hema substratum. We then investigated the effects of berberine on the growth of these cells. The anoikis-resistant cells had a reduced growth rate and were more invasive than their respective adherent cell lines. The effect of berberine on growth was compared to that of doxorubicine, which is a drug commonly used to treat breast cancer, in both the adherent and anoikis-resistant cell lines. Berberine promoted the growth inhibition of anoikis-resistant cells to a greater extent than doxorubicine treatment.*

Treatment with berberine-induced cell cycle arrest at G0/G1 in the anoikis-resistant MCF-7 and MDA-MB-231 cells as compared to untreated control cells. In summary, these results revealed that berberine can efficiently inhibit growth by inducing cell cycle arrest in anoikis-resistant MCF-7 and MDA-MB-231 cells.*

Further analysis of these phenotypes is essential for understanding the effect of berberine on anoikis-resistant breast cancer cells, which would be relevant for the therapeutic targeting of breast cancer metastasis.*

A clinical study on the short-term effect of berberine in comparison to metformin on the metabolic characteristics of women with polycystic ovary syndrome.

Wei W, Zhao H, Wang A, Sui M, et al. Eur J Endocrinol. 2012 Jan;166(1):99-105. doi: 10.1530/EJE-11-0616. Epub 2011 Oct 21.

Polycystic ovary syndrome (PCOS) is a frequent reproductive and metabolic disorder associated with insulin resistance (IR). Berberine (BBR) is an isoquinoline derivative alkaloid extracted from Chinese medicinal herbs that has been used as an insulin sensitizer. BBR may have a potential therapeutic value for PCOS. The aim of this study was to evaluate the effects of BBR in comparison to metformin (MET) on the metabolic features of women with PCOS.*

Eighty-nine subjects with PCOS and IR subjects were randomized into one of three treatment groups: BBR+compound cyproterone acetate (CPA; n=31), MET+CPA (n=30), and placebo+CPA (n=28) for 3 months. Clinical characteristics of the women and metabolic and hormonal parameters were assessed before and after the period of treatment.*

Treatment with BBR in comparison to MET showed decrease in waist circumference and waist-to-hip ratio (WHR; P<0.01), total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDLC; P<0.05) as well as increase in high-density lipoprotein cholesterol (HDLC) and sex hormone-binding globulin (SHBG; P<0.05). Similarly, treatment with BBR in comparison to placebo showed decrease in WHR, fasting plasma glucose, fasting insulin, homeostasis model assessment for IR, area under the curve of insulin, TC, LDLC, and TG (P<0.05) as well as increase in HDLC and SHBG (P<0.01).*

Intake of BBR improved some of the metabolic and hormonal derangements in a group of treated Chinese women with PCOS. Main effects could be related to the changes in body composition in obesity and dyslipidemia. Further controlled studies are needed for the assessment of the potential favorable metabolic effects of BBR in women with PCOS.*

Effect of Berberine Administration on Metabolic Syndrome, Insulin Sensitivity, and Insulin Secretion*

Karina G. Prez-Rubio, Manuel González-Ortiz, Esperanza Martínez-Abundis, José A. Robles-Cervantes, and María C. Espinel-Bermúdez.

Metabolic Syndrome and Related Disorders. June 28, 2013 doi:10.1089/met.2012.0183.

The aim of this study was to evaluate the effect of berberine administration on metabolic syndrome, insulin sensitivity, and insulin secretion.

Methods: A randomized, double-blind, placebo-controlled clinical trial was carried out in 24 patients with a diagnosis of metabolic syndrome. Glucose and insulin levels after a dextrose load were measured. Triglycerides and high-density lipoprotein cholesterol concentrations at baseline were also measured. Twelve patients received berberine hydrochloride (500‚Äâmg) three times daily before meals for 3 months. The remaining 12 patients received placebo. Area under the curve (AUC) of glucose and insulin, total insulin secretion, first-phase of insulin secretion, and insulin sensitivity were assessed.*

Results: After berberine administration, patients had a remission of 36% (P=0.037) in the presence of metabolic syndrome and a significant decrease in waist circumference in females (106±4 vs. 103±3 cm, P<0.05), systolic blood pressure (SBP) (123±7 vs. 115±9 mmHg, P<0.01), triglycerides (2.4±0.7 vs. 1.4±0.5 mmol/L, P<0.01), area under the curve (AUC) of glucose (1182.1±253.6 vs. 1069.5±172.4 mmol/l, P<0.05), AUC of insulin (92,056±72,148 vs. 67,407±46,441 pmol/L, P<0.01), and insulinogenic index (0.78±0.69 vs. 0.62±0.46, P<0.05), as well as an increase in the Matsuda index (2.1±1.0 vs. 3.1±1.6, P<0.01).*

Conclusion: Administration of berberine leads to remission of metabolic syndrome and decreases in waist circumference, SBP, triglycerides, and total insulin secretion, with an increase in insulin sensitivity.*

Berberine is Superior to Metformin*

Block, W. (2011, July). Berberine is Superior to Metformin. Life Enhancement. Retrieved from http://www.life enhancement.com/magazine/article/2439-berberine-is-superior-to-metformin.*

Metformin a widely used first-line antidiabetic drug prescribed by doctors for the treatment of type 2 diabetes, particularly in the overweight and obese. A new review focuses on several studies showing that the plant alkaloid berberine can lower blood glucose as effectively as the drug metformin at similar doses (500 mg, taken 3 times/day), and perhaps even better in some ways.[1]*

Berberine is found in Coptis chinensis (goldenthread), Berberis aquifolium (Oregon grape), Berberis vulgaris (barberry), Hydrastis canadensis (goldenseal), and Berberis aristata (tree turmeric). Traditionally, it has been used for more than 2500 years in both Ayurvedic and Chinese medicine, with growing interest in its effects in metabolic and cardiovascular disease in the Western world in the last decade. Berberine has a wide range of healthful uses that include cardiovascular, anti-inflammatory, and antimicrobial (it acts against bacterial diarrhea, intestinal parasites, fungal infections, Candida albicans, yeast, and possibly methicillin-resistant Staphylococcus aureus).*

Berberine for Diabetes Mellitus Type 2*

Recent reviews show the effectiveness of berberine compared to metformin for type 2 diabetes. [2] In the first study, 36 adults with newly diagnosed type 2 diabetes were randomly assigned to treatment with berberine or metformin (500 mg 3 times/day) in a 3-month trial. The hypoglycemic effect of berberine was similar to that of metformin. Metformin is a widely used first-line antidiabetic drug prescribed by doctors for the treatment of type 2 diabetes, particularly in the overweight and obese, and those with normal kidney function. Significant changes were observed in the berberine group:*

Hemoglobin A1c (glycosylated hemogblobin, an AGE*) decreased from 9.5% to 7.5% (about a 21% reduction)

Fasting blood glucose (FBG) decreased from 190.8 to 124.2 mg/dl

Postprandial blood glucose (PBG) decreased from 356.4 to 199.8 mg/dl

Plasma triglycerides from 100.5 to 79.2 mg/dl

* Advanced glycation end-products (AGEs) result from a chain of chemical reactions after an initial glycation reaction. The intermediate products are known, variously, as Amadori, Schiff base, and Maillard products (named after the researchers who first described them). Side products generated in intermediate steps may be oxidizing agents (such as hydrogen peroxide), or not (such as beta amyloid proteins). "Glycosylation" is sometimes used for "glycation" in the literature, usually as "non-enzymatic glycosylation."*

In the second study, 48 adults with poorly controlled type 2 diabetes were supplemented with berberine in a 3-month trial:*

Hemoglobin A1c decreased from 8.1% to 7.3% (about a 10% reduction)

In the first 7 days of treatment, berberine led to a reduction in FBG from 172.8 to 140.4 mg/dl and PBG from 266.4 to 210.6 mg/dl

During the second week, FBG and PBG declined further, reaching a low point that was 38 mg/dl below the baseline of 135 mg/dl for FBG and 60 mg/dl below the baseline of 189 mg/dl for PBG, and remained at these levels thereafter

Fasting plasma insulin was reduced by 28.1%

The homeostasis model assessment of insulin resistance index was reduced by 44.7%

Total cholesterol and low-density lipoprotein cholesterol were decreased significantly

Of further interest, both fasting and postprandial proinsulin C-peptides increased significantly in patients when berberine was used together with insulin. These peptides facilitate the efficient assembly, folding, and processing of insulin in the endoplasmic reticulum, and their increase suggests that long-term berberine treatment may improve insulin secretion of the patients. During the trial, 20 patients experienced transient gastrointestinal adverse effects. Functional liver or kidney damages were not observed in any patients.*

Berberine Decreases Cholesterol and Triglyceride*

Another recent study, covered in the review, showed that berberine benefits type 2 diabetes.3 In this study, 116 patients with type 2 diabetes and dyslipidemia were randomly allocated to receive berberine (1 g daily) or placebo for 3 months. In the berberine group:*

Hemoglobin A1c decreased from 7.5% to 6.6% (about a 12% reduction)

FBG decreased from 7.0 to 5.6 mm/L (126 to 100.8 mg/dl)

PBG decreased from 12.0 to 8.9 mm/L (216 to 160.2 mg/dl)

Triglycerides decreased from 2.51 to 1.61 mm/L (220 to 141 mg/dl)

Total cholesterol decreased from 5.31 to 4.35 mm/L (205 to 168 mg/dl)

LDL cholesterol decreased from 3.23 to 2.55 mm/L (124.9 to 98.6 mg/dl)

The glucose disposal rate was increased after berberine treatment, although no significant change was found between berberine and placebo groups. Mild to moderate constipation was observed in 5 participants in the berberine group. Transient gastrointestinal adverse effects with berberine were fairly common and may be related to its antimicrobial action. Berberine may be particularly useful in cases involving both type 2 diabetes, dyslipidemia, and possibly infection.*

Berberine Offers Antiobesity Effects

Obesity is a major cause of metabolic syndrome and is due to an increase in the number and hypertrophy (volume increase) of adipocytes. But if the differentiation and proliferation of adipocytes is inhibited, then metabolic syndrome may be treated and prevented. A new study investigated the effects of 50 commonly used Kampo (the Japanese study and adaptation of Traditional Chinese medicine) preparations on the differentiation of 3T3-L1 preadipocytes to search for a drug with an antiobesity effect. [4]*

Kampo medicines were screened, and the strongest differentiation-inhibitory effect was noted with Orengedokuto (OGT). To explore the active ingredients in OGT, the effects of four crude components of OGT were investigated, and it was found that the differentiation-inhibitory effects of OGT was accounted for by Coptidis rhizome and Phellodendri cortex, both of which contain berberine, which upon examination also showed a differentiation inhibitory effect.*

In fact, berberine was found to inhibit the mRNA and protein expression of PPARγ as well as the CCAAT/enhancer binding protein α (C/EBPα).* Moreover, berberine inhibited lipid accumulation in adipocytes. These findings suggest that an antiobesity effect could be a new indication for OGT and that its active ingredient is berberine, with a mechanism involving the inhibition of PPARγ and C/EBPα expression. This means that berberine could reduce the size of your fat cells and cut down on their number as well.*

* C/EBPα is a member of a family of transcription factors, composed of six members. They promote the expression of certain genes through interaction with their promoter. C/EBPα is required for both adipogenesis (fat cell creation) and normal adipocyte (fat cell) function.*

Berberine for Memory Too

We should not forget that berberine has also recently been found to enhance memory function in rats. In research done in India, berberine is reported to inhibit cholinesterase (ChE) activity and increase glucagon-like peptide (GLP-1) release. [5] ChE is the enzyme that breaks down the memory molecule acetylcholine, a neurotransmitter that is crucial for the important memory activities of focus and concentration. GLP-1, as recent evidence suggests, plays an imperative role in diabetes, along with cognitive dysfunction, learning, and neuroprotection.*

Based on the unavailability of prior research for the influence of berberine on streptozotocin (STZ)-induced memory impairment, the researchers designed their study to investigate if there could be any benefits. The researchers used the Morris water maze model as a measure of memory. In the Morris test, rats are trained to find a submerged platform in a tank of water. Around the sides of the tank are symbols, and the goal entails (pun intended) that the rats remember the location of the platform, to save themselves from exerting energy to stay afloat by refinding the platform and emerging from the water.*

Also, lipid peroxidation and glutathione levels (as parameters of oxidative stress) and ChE activity (as a marker of cholinergic function) were assessed in the cerebral cortex and hippocampus. The greater the activity of ChE, the less effective is the cholinergic system, as ChE breaks down acetylcholine. Thirty days after diabetes induction with STZ, rats showed a severe deficit in learning and memory associated with increased lipid peroxidation, decreased reduced glutathione, and elevated ChE activity.*

In contrast, diabetic rats treated chronically with orally administered berberine at doses of 25, 50, or 100 mg/kg, but especially the 50 and 100 mg dose, twice daily for 30 days, and then subjected to the Morris water maze test, were all found to have improved cognitive performance, along with lowered hyperglycemia, oxidative stress, and ChE activity. Also administered was vitamin C (100 mg/kg) or metformin (500 mg/kg) or vehicle (1 ml/kg) as placebo, twice daily. Both vitamin C and metformin were comparable in effects to berberine.*

Diabetes I in Cognitive Dysfunction

Prior studies have identified that when memory impairment is induced in rats, changes consequently occur in the central nervous system that are secondary to hyperglycemia, including impaired oxidative stress, cholinergic dysfunction, and changes in GLP-1. Indeed, in a model of treatment that uses antihyperglycemics, antioxidants, and cholinergic agonists, beneficial effects have been produced.*

In the second leg of the Indian study, 30 days after confirmation of diabetes, berberine was given at the same dose range given during training trials in the Morris water maze that lasted 5 days (days 31-35). Alternatively, vitamin C at 100 mg/kg or metformin at 500 mg/kg, or donepezil (an AChEI that is similar to the plant nutrient galantamine) at 3 mg/kg or placebo at 1 ml/kg were also used. The result for berberine ingestion was improved learning and memory, with lowered hyperglycemia, oxidative stress, and ChE activity. Chronic treatment (30 days) with vitamin C, or metformin, or donepezil during training trials also improved diabetes-induced memory impairment and reduced oxidative stress and/or cholinesterase activity. In a prior study with berberine, its effects were found to be very similar to metformin, and this was upheld in the current study.*

Significanat Effect on Diabetes

Berberine has been shown to have a significant beneficial effect on type 2 diabetes, and may be as effective, or more so, than metformin. Berberine acts through several mechanisms, including mimicking insulin; improving insulin action by activating AMPK; reducing insulin resistance through protein kinase C-dependent up-regulation of insulin receptor expression; inducing glycolysis; and on incretins by promoting GLP-1 secretion and modulating its release, and by inhibiting DPP-4.*

Berberine's Probable and Possible Mechanisms of Action

The studies encompassed by the review that is the central topic of the article "Berberine is Superior to Metformin" (of which this sidebar is a part) have found that berberine is beneficial for glucose metabolism and insulin activity through a number of distinct mechanisms:*

Mimicking

Berberine mimics insulin action by increasing glucose uptake ability by 3T3-L1 adipocytes (fat cells) and L6 myocytes (muscle cells) in an insulin-independent manner. Furthering this mimicking process, berberine inhibits activity of protein tyrosine phosphatase 1B (an important negative regulator of insulin and leptin signaling in vivo). It also increases phosphorylation in 3T3-L1 adipocytes. In diabetic mice, berberine lowers hyperglycemia and improves impaired glucose tolerance, but does not increase insulin release and synthesis. The results suggest that berberine represents a different class of anti-hyperglycemic agents.*

Improving

By activating AMPK [2-5] AMPK (AMP-activated protein kinase) is an enzyme that plays a role in cellular energy homeostasis. Principally, the effect of AMPK activation is stimulation of hepatic fatty acid oxidation and ketogenesis,* inhibition of cholesterol synthesis, lipogenesis (the formation of fat), and triglyceride synthesis, inhibition of adipocyte lipolysis and lipogenesis, stimulation of skeletal muscle fatty acid oxidation and muscle glucose uptake, and modulation of insulin secretion by pancreatic beta-cells. [6]*

AMPK is expressed in a number of tissues, including the liver, brain, and skeletal muscle, where it acts as a "metabolic master switch" that regulates several intracellular systems, including the cellular uptake of glucose, the beta-oxidation of fatty acids, and the biogenesis of glucose transporter 4 (GLUT4).[7]*

* Ketogenesis is the process by which ketone bodies are produced as a result of fatty acid breakdown. Glucose is usually used by cells for energy. But, when there's no insulin to help it transport out of the blood and into the cells, the body has an "energy crisis" and starts to break down body fat into ketones as an alternative fuel source. This is called ketosis.*

Reducing resistance through protein kinase C (PKC)-dependent up-regulation of insulin receptor (InsR) expression [8,9]Berberine induced InsR gene expression through a PKC-dependent activation of its promoter. Inhibition of PKC abolished berberine-caused InsR promoter activation and InsR mRNA transcription. In animal models, treatment of type 2 diabetes mellitus rats with berberine lowered fasting blood glucose and fasting serum insulin, increased insulin sensitivity, and elevated InsR mRNA as well as PKC activity in the liver.*

Berberine has recently been reported to activate AMPK and increase its phosphorylation, which was associated with persistent elevation in AMP/ATP ratio and reduction in oxygen consumption. An increase in glycolysis was observed with a rise in lactic acid production. The results of this suggest that berberine enhances glucose metabolism by stimulation of glycolysis, which is related to inhibition of glucose oxidation in mitochondria. Berberine-induced AMPK activation is likely a consequence of mitochondria inhibition that increases the AMP/ATP ratio.*

Promoting

etion and modulating its release [11,12]Glucagon-like peptide (GLP)-1 is a potent glucose-dependent insulinotropic gut hormone released from intestinal L cells. GLP-1 is one of the incretins, a group of gastrointestinal hormones that cause a short-term increase in the amount of insulin released from the beta cells of the Islets of Langerhans (which also produce insulin in the pancreas) after eating (see above image). Several approved drugs act on incretins, but there are many side effects, a significant number of which are gastrointestinal. This is ironic because berberine has been used to treat gastrointestinal problems.*

Berberine increased GLP-1 secretion in streptozotocin-induced diabetic rats. In vivo, 5-week treatment of berberine enhanced GLP-1 secretion induced by glucose load and promoted proglucagon mRNA expression as well as L cell proliferation in intestine. In vitro, berberine concentration-dependently stimulated GLP-1 release in NCI-H716 cells. Berberine also promoted both prohormone convertase 3 and proglucagon mRNA expression. This demonstrates that berberine showed its modulation on GLP-1 via promoting GLP-1 secretion and GLP-1 biosynthesis. *

Inhibition

[13]Berberine was investigated as an inhibitor of human dipeptidyl peptidase IV (DPP IV) in an attempt to explain its anti-hyperglycemic activities. The investigation included simulated docking experiments to fit berberine within the binding pocket of DPP IV. Berberine was found to readily fit within the binding pocket of DPP IV in a low energy orientation characterized with optimal electrostatic attractive interactions bridging the isoquinolinium positively charged nitrogen atom (berberine) and the negatively charged acidic residue of glutamic acid-205 (GLU205) of DPP IV.*

Experimentally, berberine was found to inhibit human recombinant DPP IV in vitro with IC(50) = 13.3 microM. These findings suggest that DPP IV inhibition is, at least, one of the mechanisms that explain the anti-hyperglycemic activity of berberine. The fact that berberine was recently reported to potently inhibit the pro-diabetic target human protein tyrosine phosphatase 1B (h-PTP 1B) discloses a novel dual natural h-PTP 1B/DPP IV inhibitor. *

Inhibition

c gluconeogenesis [14]Berberine is a compound originally identified in a Chinese herbal medicine Huanglian (Coptis chinensis French). It improves glucose metabolism in type 2 diabetic patients. The mechanisms involve in activation of adenosine monophosphate activated protein kinase (AMPK) and improvement of insulin sensitivity. However, it is not clear if berberine reduces blood glucose through other mechanism. In this study, we addressed this issue by examining liver response to berberine in diabetic rats, in which hyperglycemia was induced in Sprague-Dawley rats by high fat diet. We observed that berberine decreased fasting glucose significantly. Gluconeogenic genes, Phosphoenolpyruvate carboxykinase (PEPCK) and Glucose-6-phosphatase (G6Pase), were decreased in liver by berberine. Hepatic steatosis was also reduced by berberine and expression of fatty acid synthase (FAS) was inhibited in liver.*

Activities of transcription factors including Forkhead transcription factor O1 (FoxO1), sterol regulatory element-binding protein 1c (SREBP1) and carbohydrate responsive element-binding protein (ChREBP) were decreased. Insulin signaling pathway was not altered in the liver. In cultured hepatocytes, berberine inhibited oxygen consumption and reduced intracellular adenosine triphosphate (ATP) level.*

The data suggest that berberine improves fasting blood glucose by direct inhibition of gluconeogenesis in liver. This activity is not dependent on insulin action. The gluconeogenic inhibition is likely a result of mitochondria inhibition by BBR. The observation supports that berberine improves glucose metabolism through an insulin-independent pathway.*

Cytotoxicity of berberine on human cervical carcinoma HeLa cells through mitochondria, death receptor and MAPK pathways, and in-silico drug-target prediction.

Lu B, Hu M, Liu K, Peng J. Toxicol In Vitro. 2010 Sep;24(6):1482-90. doi: 10.1016/j.tiv.2010.07.017. Epub 2010 Jul 23.

Berberine, a natural product, has been widely used to treat hyperlipoidemia and intestinal diseases. In the present paper, berberine showed a significant anti-proliferative effect to human cervical carcinoma HeLa cells confirmed by 3-(4,5)-dimethyl-thiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT), flow cytometry analysis (FCM) and so on.*

The methods including western blotting, radioimmunity assay (RIA), reverse transcription-polymerase chain reaction (RT-PCR) were used to investigate protein and mRNA expressions. We found that Bcl-2/Bax ratio was significantly decreased and cytochrome c was released from mitochondrion to cytosol, which indicated that the mitochondrial pathway was activated by berberine. The up-regulation of Fas, FasL, TNF-alpha and TRAF-1 indicated the involvement of the death receptor pathway in the process of berberine-induced apoptosis.*

Furthermore caspase-3 and caspase-8 were activated as a central event of apoptosis, and the levels of phosphorylation of mitogen-activated protein kinases (MAPKs) were also investigated. In addition, the increased expression of p53 was also observed in berberine-treated HeLa cells, and as a node point of these different pathways in a protein-protein interaction network constructed by GeneGo software, p53 might be the possible drug-target of berberine's anti-cancer on HeLa cells, which was predicted by a flexible ligand-protein inverse docking program, INVDOCK. This study is benefit for clarifying the mechanism of berberine's anti-tumor effect and might be helpful to find therapy-target for treatment of human cervical carcinoma.*

Rhizoma Coptidis was first reported in Shen non's Herbal (Shen Nong Ben Cao Jing), and has the effect of clearing heat and drying dampness, purging fire to eliminate toxin. Berberine (BBR) is the main alkaloid of Coptis Chinensis. It is widely used in Chinese medicine (CM) as an antimicrobial for treating dysentery and infectious diarrhoea (Yang & Lin, 2007). Recent researches showed that BBR had the effects of antiarhythmia, expanding coronary artery, regulating blood lipids, blood pressure, blood glucose, etc (Cheng Y& Zhong, 2007). Presently, He, et al (2006) reported that BBR could inhibit the formation of carotid atherosclerosis (AS). As a natural drug, it has the effect of inhibiting atherosclerosis through multiple links with few side effects, and is suitable for long time use. The mechanism of BBR for anti-atherosclerosis was reviewed from regulating lipids, anti-inflammation, decompression, reducing blood sugar, and inhibiting proliferation of vascular smooth muscle cells (VSMCs).*

Lipid infiltration is one of the mechanisms of AS (Li, 2001). The atherosclerotic process is initiated when cholesterol-containing low-density lipoprotein (LDL) accumulates in the intima and activates the endothelium(Hansson et al, 2006). Inflammatory reaction caused by lipid infiltration is an important pathological process. It has been clinically proved that increased total cholesterol (TC) played an important role in AS, and LDL-C played fundamental effects. Oxidative LDL (ox-LDL) is correlated with plaque formation and stability (Griendling & Alexander, 1997).*

High densitylipoprotein cholesterol (HDL-C) has the effect of anti-atherosclerosis through transferring TC from extrahepatic tissues to liver (Crouse et al 1995). In the present study, BBR was found to display antiproliferative action against VSMC by blocking the cell cycle at G0 and G1/M phases. The mechanism underlying the G1 arrests involved a decrease in cyclin D1 protein (Tanabe et al, 2005). Further results showed that BBR could decrease the growth factor significantly, mainly angiotensinⅡ (Ang Ⅱ) and heparin binding epidermal growth factor (HB-EGF), to inhibit VSMC proliferation and migration in vitro, and this effect was achieved by delaying or partially suppressing activation of Akt pathway rather than ERK pathway (Lee et al, 2006). However, recent studies showed that BBR blocked injury-induced SMC re-growth by inactivation of ERK/Egr-1 signalling pathway and downstream target (Egr-1, c-Fos, cyclin D1 and PDGF-A) expression (Liang et al, 2006). In addition, BBR could exert anti-atherosclerotic effect through inhibiting platelet aggregation (Huang et al, 2002), resisting lipid peroxidation (Jin et al, 2000) and regulating apoptosis (Fang et al, 2002).*