The Effect Of Berberine On The High Androgen State And The Expression Of St AR’s MRNA And Protein In PCOS Rats

Objective Guided by the theories of “PCOS clinical manifestations belongs to damp-heat syndrome”in TCM and “insulin resistance is PCOS basic pathological change” in Western medicine, with “berberine heat-clearing and damp-drying, improving insulin resistance” as the breakthrough point, the effect and mechanism of berberine in treating PCOS were analyzed and summarized. The effects of berberine on weight,testosterone,fasting insulin,blood glucose and sex hormone-binding globulin in PCOS rats were systematically detected through animal experiment. The effects of berberine reducing polycystic ovary and ovulation induction in PCOS rats were observed. Whether berberine could treat PCOS was evaluated. The change of St AR in Berberine treating PCOS rats was analyzed. The mechanism of berberine to treat PCOS was investigated.Method1.Modeling method: 48 SPF female SD rats(6 weeks) were fed with the pellet feed and free water. After adaptive feeding for 3d, 48 rats were randomly divided into two groups: normal control group(N=12) and model group(N=36). The rats in normal control group were intragastrically administrated with 1%carboxymethylcellulose for continuous 21 d;The rats in model group were intragastrically administrated with 1mg/kg letrozole +1%carboxymethylcellulose for continuous 21 d. The PCOS rat model induced by letrozole was established.2.Grouping and treatment method The rat models were not successfully made in group A withdrawal from the experiment. The successful model rats were randomly divided into Model group, metformin group, and berberine group.After the end of the model, the medicine was administered in rats.The administration plan was as follows:A(Blank control group): N=12, No modeling, According to4ml/kg/d drenching 1% carboxymethyl cellulose in 3 consecutive weeks;B(Model control group): N=12, After modeling, According to4ml/kg/d drenching 1% carboxymethyl cellulose in 3 consecutive weeks;C(metformin group):N=12,After modeling,Drenching by filling1% of metformin into carboxymethyl cellulose with 4ml/kg/d.According to ratio table of body surface area between human and animal,the routine dose of metformin in treating PCOS was 500 mg for 3 times a day taken orally. After conversion, the given dose for rats was 135mg/kg/d for 3 weeks.D(berberine group):N=12,After modeling,Drenching by filling berberine into 0.9% normal saline with 4ml/kg/d. According to ratio table of body surface area between human and animal, the common dose of berberine for people was 0.3g each time for 3 times a day.After conversion,the given dose for rats was 81mg/kg/d for 3 weeks.3. Observe indicatrix3.1 Observe the variation of rats’ weight and serum testosterone level of each group before and after treatment.3.2 Observe the difference between groups in fasting insulin,fasting blood glucose, and sex hormone binding globulin after treatment.3.3 Observe the rat ovarian morphological difference among groups after treatment.3.4 Observe the key factors of synthetizing androgen, St AR protein and m RNA expression, in rat ovarian among groups via immunohistochemical and PCR technique.Rusult1.modeling results The typical PCOS manifestations in this experiment of the model rats induced by Letrozole were:Vaginal shedding cell smear loss of regularity of dynamic cycle,vaginal epithelial cells were stuck in the early stages of the follicular phase, suggestingof no ovulation; The pathological results showed that the ovary of the rat was changed with polycystic;serum testosterone elevated;Weight increased; Fasting insulin was increased, which indicated the existence of insulin resistance.The PCOS rat model was successfully produced by using the Letrozole.2. Weight Letrozole induced PCOS rats showed remarkable weight gain(in contrast with the blank comparison group, P<0.05). Both metformin and berberine could control the weight gain of PCOS rats(in contrast with the model comparison group, P<0.05), but they failed to return the rats` weight to normal standard(in contrast with the blank comparison group, P<0.05).3. Testosterone Letrozole induced PCOS rats showed remarkable increase oftestosterone level(in contrast with the blank comparison group,P<0.05). Metformin could not decrease Testosterone level of PCOS rats(in contrast with the model comparison group, P>0.05).berberine could decrease testosterone level of PCOS rats(in contrast with the model comparison group, P<0.05), but it failed to return the testosterone level of PCOS rats to normal standard(in contrast with the blank comparison group, P>0.05).4. Fasting insulin Letrozole induced PCOS rats showed remarkable increase in fasting insulin level(in contrast with the blank comparison group,P<0.05). Both metformin and berberine could return the fasting insulin level of PCOS rats to normal(in contrast with the blank comparison group, P>0.05).5. Fasting blood-glucose Fasting blood-glucose induced GLU level of PCOS rats were normal(in contrast with the blank comparison group P>0.05).6. Sex hormone binding globulin The PCOS rat models with Letrozole in this study showed increase of SHBG(in contrast with the blank group P<0.05), and the level of SHBG had positive relationship with the value of testosterone.7. Pathology of ovarian under light microscope Letrozole induced PCOS rats` ovarian were enlarged, the surface obvious pale, outer ovariolar sheath thickening, ovarian multiple follicular cysts,follicular theca cells thickening,and no corpora luteum was seen in the ovarian, prompting no ovulation. After treatment, the group using Metformin still had above pathogenic manifestations, showing no obvious difference with the model comparison group. The group treated with berberine showed decreasein ovarian volume and the number of ovarian multiple follicular cysts, thinning of outer ovariolar sheath. No follicular cystic change was observed and multiple corpora luteum were showed in the ovarian, prompting oligomenorrhea ovulation.8.St AR Immunohistochemistry results showed that Letrozole induced PCOS rats had remarkable increase in St AR density on the thecal cell membrane(in contrast with the blank comparison group P<0.05).The St AR density of the metformin group showed no apparent difference with the model comparison group(p>0.05). The St AR density of the berberine group was lower than those of the metformin and model comparison group(P<0.05), but larger than that of the blank comparison group(P<0.05). It was found that the St AR protein expression level was positively related to the value of testosterone.The analysis of the PCR result showed that the m RNA expression level of St AR in the ovarian was positively related to the St AR protein density, which manifested that the St AR could not only be over-expressed in ovarian tissue, but also synthesized in ovarian in large scale.Conclusion Testosterone levels in PCOS Kaohsiung were positively correlated with St AR density in the cell membrane of the follicular cells.Metformin in the treatment of PCOS can only improve insulin resistance and lower body weight. While the berberine in the treatment of PCOS can not only improve insulin resistance, reduce weight, but also the treatment of hormone in Kaohsiung. Berberine can reduce the theca cell through the cell membrane on the densityof St AR to reduce the PCOS value of testosterone.So the conclusion that we come to was: Berberine can treat PCOS,and it not only can improve insulin resistance, but also treat Kaohsiung hormone syndrome.