The Study Of Improving Rat Polycystic Ovarian Syndrome-Insulin Resistance (PCOS-IR) Model And The Influence Of The Jia Wei Cang Fu Dao Tan Tang Decoction On The Model

ObjectiveThe study is based in the traditional PCOS rat model induced by Dehydroeplandrosterone(DHEA),to establish the improved Puberty PCOS-IR rate model.While used the Chinese Herbal Jia Wei Cang Fu Dao Tan Tang as the treatment,to investigate the effect on weight,organ weight,hormonal, metabolized and morphology changes in the ovary of PCOS rat model.In Addition, the study could explore the mechanism of action towards reproductive system after using a herbal formula for supplementing the kidney and lustrating Tan.MethodExperiment 1:Establishment of improved Puberty PCOS-IR rate model52 21^st day SPF SD female rats which weight controlled between 35-45g were randomized into five groups.Group A(Sodium Prasterone Sulfate of injection shorted for T):Begin in 23^rd day,hypodermic injection with Sodium Prasterone Sulfate of injection 9mg/100g(equal to DHEA6mg/100g ) for 40 days.Group B(T+HCG):The same as group A,and from 27^th day hypodermic injection with HCG 1.5u twice a day for 35 days.Group C(HCG):From 23^th day hypodermic injection with HCG 1.5u twice a day for 40 days.Group D(Blank control group):From 23^th day hypodermic injection with Physiological saline 0.2ml for 40 days.Group E(High Fat Feed group):The same as group D.Feed adaptively on normal feedstuff for 2 days.On 23rd day,except Group D only feed on normal feedstuff for 40 days,the other groups will be feed on normal feedstuff for the first 20 days and High Fat Feedstuff for the late 20 days.To observe the general condition such as weight,eating,excrement,Vagina smear and so on.Fasting but not endure the water on the 40^th day night,all rats were killed the next day.Anaesthesia by 10%Chloral hydrate,draws blood from the vein of eye socket,conserved the blood serum to test hormonal and metabolized index;weight ovary,uterus and fat around reproductive organ in by electronic balance.The histological and morphological alteration of ovarian were studied with microscope.Ovary ultrastrusture was detected by electric microscope.Experiment 2:The Influence of the Jia Wei Cang Fu Dao Tan Tang Decoction on PCOS-IR Rat ModelJia Wei Cang Fu Dao Tan Tang Decoction were divided into 2 parts—Wen Shen Jian Pi Decoction and Li Qi Hua Tan Decoction.90 21^st day SPF SD female rats were randomized into 9 groups.All groups will be orally fed for 21 days and have to change dosage when rats weight gained,group A were treated with Jia Wei Cang Fu Dao Tan Tang Decoction at 48.4g/kg/d.Group B Jia Wei Cang Fu Dao Tan Tang Decoction at 12.1g/kg/d.Group C were treated with Wen Shen Jian Pi Decoction at 23.6g/kg/d.Group D were treated with Wen Shen Jian Pi Decoction at 5.9g/kg/d.Group E were treated with Li Qi Hua Tan Decoction at 24.8g/kg/d. Group F were treated with Li Qi Hua Tan Decoction at 6.2g/kg/d.Group G were treated with Metformin at 0.19g/kg/d.Group H(model group) and GroupⅠ(High Fat Feed group) were treated with Physiological saline.The establishment of rat model and the observation projects were the same as experiment 1 group B.Result1.This experimental rat model has following characteristic:(1) Body weight:,Although there was no significant difference between the model group and the blank group,but compared with the blank group,the weight of rats in the three kind of PCOS models and High Fat Feed group grown more quickly.(2) Fat agglomeration around ovary and reproductive organ:Used androgen only would cause the ovary volume of rat model too small,even smaller than the blank control group;and it has not appeared ovary sclera accumulation as human PCOS.But if added with HCG may increases obviously the volume and the wet weight of ovary.Compared with the blank group,the T+HCG group and the HCG group ovary wet weight has statistics significance(p＜0.05).The fat of the T+HCG group around the reproductive organ was significantly heavier than the blank group(p＜0.05).The application of T+HCG union may enlarge the effect which the high fat feed could increase the weight,and changed the bodily fat distribution,causes the abdomen fat increase to be obvious,in particular around reproductive organ fat increase.(3) Endocrine changes:The androgen and the androstenedione of the T+HCG group were significantly higher than the blank group(p＜0.05).The application of T and HCG union may enlarge the effect of the androgen and increase androgen biological exploitability.Also elevates the androstenedione primarily.(4) Metabolism changes:PCOS characteristic is obvious in the T+HCG group. Application of the T+HCG union may cause the insulin remarkable ascension, compared with the blank group it has statistics significance(p＜0.05).It infers that the high insulin blood syndrome is the important link of PCOS pathology process.HCG united the high fat feed could cause unusual metabolism, and took the cholesterol markup as the characteristic.(5) Morphology changes:Draw the conclusion from the statistical analysis, three kinds of PCOS models may achieve that the ovulation had been suppressed. It showed that the main function of the androgen lied in the suppression to the ovulation in morphology change,added with HCG may enlarge the effect of the multi-pouches change of the model.(6) The pure high fat feed does not cause the increase of the ovary wet weight and the multi-pouches changed,also does not cause the endocrine hormone obvious changed.The application of Sodium Prasterone Sulfate of injection +HCG and the high fat diet union,may play the supplementary role,caused the performance of animal model much closer to clinical PCOS in the endocrine,metabolism and the morphology changes.2.The Influence of the Jia Wei Cang Fu Dao Tan Tang Decoction on PCOS-IR Rat Model(1) Body weight and fat agglomeration around ovary and reproductive organ: The Jia Wei Cang Fu Dao Tan Tang Decoction has no significance influence on the body of the rat model(P＞0.05).It showed that other methods such as sports and diet control would be a good intervention on over-weight problem of PCOS. In the experiment 1,compared with the blank group,the fat around the reproductive organ of T group,T+HCG group and the HCG group were significantly heavier(p＜0.05);while in the experiment 2,although there were no significantly difference in each group(P＞0.05),but compared with the model group,the fat around the reproductive organ of each group reduced.It showed that there were certain significance of decoction intervention ahead of time which could adjust the abdomen fat in the distribution and the fat cell factor metabolism.The decoction did not have the influence regarding to the wet weight of uterus.(2) Wet weight of ovary:Compared with the blank group,the ovary weight of other groups were significantly lighter(p＜0.05).There is two reasons:First, because of decoction intervention ahead of time,it may had the prevention and the treatment dual function;Second,it possibly promoted the ovulation, and caused the liquid discharged from the pouch of the ovary,thus promoted ovary volume and weight decline.(3) Endocrine changes:①The content of androsterone in high dose groups were significantly lower than the low dose groups(p＜0.05),Chinese herbal had certain dose-effect relationship.②There were no obvious rule in decreasing LH.Instead of reducing but elevating LH,whether LiQiHuaTan group with acceleration of the ovulation,induced the LH peak or not still under confirmation.For the whole,high dose of JiaWeiCangFuDaoTanTang group was the strongest in decreasing the endocrine hormone.The superiority function of coordination treatment of Chinese herb was conformed.(4) Metabolism changes:Metformin enhanced the insulin sensitivity.Insulin in high does of LiQiHuaTan group was lower than the metgormin group,both two groups were significantly lower than the blank group(p＜0.05).It considered that it was more effective for LiQiHuaTan herb in reducing insulin and enhances the excitability of insulin.The glycerin had no significantly change in experiment 1 and 2(P＞0.05).It may be considered that the high fat model did not cause the glycerin territory value change by no means. JiaWeiCangFuDaoTanTang decoction,LiQiHuaTan decoction and metgormin were more effective in decreasing cholesterol than WenSenJianPi decoction,it showed that WenSenJianPi decoction could possibly in the adjustment organism metabolism,in particular falling tone cholesterol.(5) Morphology changes:The quantity of bursa ovarica change,the mature graafian follicle and the corpora luteum were not obvious,the curative treatment effect of Chinese native medicine mainly manifests was promoting the function of pouch expansion graafian follicle restoration----Namely promoting the granular cell layer increased,oviferous tubercle and transparent belt reproduced and the corpora luteum matured and so on. JiafeiCangFuDaoTanTang decoction and WenSenJianPi decoction were helpful in promoting the pouch graafian follicle function to restore,to press the ovulation and to perfect and maintain the function of the yellow body.ConclusionWhile used the Chinese Herbal Jia Wei Cang Fu Dao Tan Tang as the treatment, to investigate.In Addition,the study could after using a herbal formula for supplementing the kidney and lustrating Tan.This research successfully improved the puberty PCOS-IR rat model. Through the observation of the effect on weight,organ weight,hormonal, metabolized and morphology changes in the ovary of PCOS rat model,we concluded that JiaWeiCangFuDaoTanTang decoction could explore the mechanism of action towards reproductive system.1.The improved puberty PCOS-IR rat model has following characteristic:Retained the high androgen syndrome and/or the high LH syndrome as the endocrine characteristics;Simultaneously it had sugar fat metabolism unusual, such as the insulin resistance,the redemptive high insulin blood sickness as well as cholesterol ascension.Has the abdomen obese characteristic----In particular fat pad which around reproductive organ obvious increased;The ovary increased obviously,Ovarian cortex accumulated,the number in ovary pouch expansion graafian follicle were remarkable increased, ovulation suppression.The ratio of PCOS model reached as high as 100%.2.JiaWeiCangFuDaoTanTang decoction could possibly improve the unusual of internal secretion and metabolism of the puberty PCOS-IR rat model.The high does of JiaWeiCangFuDaoTanTang decoction was prominent effectively in the falling tone androgen;LiQiHuaTan component was good at adjusting organism metabolism,the falling tone insulin and the cholesterol obviously;Possibly the key aspect was improved the environment of ovary and promoted ovulation;3.The high does of JiaWeiCangFuDaoTanTang decoction obviously surpasses in the synthesis curative effect compared with the other two components,it conformed that the superiority function of coordination treatment of Chinese herb.The high and low does group of Chinese herbal had certain dose-effect relationship; 4.The curative treatment effect of JiaWeiCangFuDaoTanTang decoction mainly manifested that it could promote the function of pouch expansion graafian follicle restoration----Namely promoting the granular cell layer increased, oviferous tubercle and transparent belt reproduced and the corpora luteum matured.The decoction could gradually reduce and repair abnormal changes in the ultrastructure of ovarian grafts;5.It showed that there were certain significance of decoction intervention ahead of time which could adjust the abdomen fat in the distribution and the fat cell factor metabolism.