| Objective:Because of the ambiguity of etiology and pathogenesis of polyeystic ovary syndrome(PCOS)and the heterogeneity of disease manifestations,traditional Chinese medicine has unique advantages in the treatment of this disease.It is suggested that the treatment of PCOS patients in puberty should be managed and intervened.Due to the majority of PCOS patients in puberty,most of them have symptoms of kidney deficiency and phlegm dampness such as acne,obesity and oligomenorrhea.So this research made PCOS rat model to observe the effect of BuShen Ditan Decoction on rat sex hormone level and insulin resistance and lipid metabolism,as well as the changes of miRNA expressions in rat ovarian tissue before and after modeling and drug intervention.The effect of BuShen Ditan Decoction on hormone level and glycolipid metabolism in patients with PCOS and insulin resistance in puberty was observed in the clinical study.Methods:In the experimental study,84 rats were fed first,including the normal control group(12 rats)and the PCOS model group(72 rats).In the normal control group,the are fed with basal diet,free eating and drinking water;the model group are fed with high fat diet after referring to the Poretsky modeling method.In the first ten days,we injected insulin(dose increased from 0.5iu to 5.0iu day by day)in the medial side of thigh;from day 11,we use 5.0iu insulin qd.and HCG(1.5iu dissolved in 0.2ml saline)bid subcutaneously.After the rats lost normal estrous cycles,we chose 10rats from normal group,60rats from model group,and devided these 60 rats into 5 groups:B model group,C metformin group,D diane-35 group,E BuShen DiTan Decoction low dose group,F BuShen DiTan Decoction middle dose group,G BuShen DiTan Decoction high dose group.The level of sex hormone,insulin resistance and lipid metabolism were compared before and after treatment.The differential expression of miRNAs in the rat ovarian tissue and the changes of miRNAs in each group were observed.In Clinical Study:90 patients with PCOS and IR in puberty were divided into three groups:Chinese medicine group,western medicine group and Chinese and Western medicine group,30 cases in each group.The Chinese medicine group was given BuShen DiTan Decoction,1 doses per day,1 courses for 3 months,2 courses of continuous treatment(except of menstrual period).The western medicine group was given metformin 500mg 2 times a day(with a meal).Integrative Medicine Group use BuShen DiTan Decoction and metformin at the same time,refering to the method of chinese medicine group and western medicine group separately.Then,we compared sex hormone levels(T、SHBG、E2、LH、FSH、PRL),free testosterone index(FAI)and insulin resistance index(IR),body mass index(BMI),waist hip ratio(WHR),ultrasonic detection and other indicators before and after treatment,and evaluated the differences of TCM syndrome integral and overall curative effect among three groups.Results:(1)The model group rats lost normal estrous cycle;compared with normal control group,the model group rats have larger ovary in pale,thicken capsular,and more subcapsular cystic follicles;the level of serum T in the rats of model group were significantly higher than the normal control group,the difference was significant(P<0.01);index of insulin resistance(HOMA index)was significantly higher than the normal control group,the difference was significant(P<0.01).(2)The weights of the normal group rats fed by standard diet were significantly different from the high fat diet rats(*P<0.05);other drug intervention group and normal control group weight have no difference from the high fat diet rats;the model group T,LH,E2 concentration was obviously higher than normal control group(P<0.05);the level of T in Chinese Medicine Groups and Diane-35 group were significantly lower than metformin group(P<0.05),indicating that Chinese Medicine and Darin-35 have better effect on reducing T.The HOMA-IR index of rats in the model group was significantly higher than that in the normal control group,indicating that the rats had insulin resistance after the model.the HOMA-IR index in Metformin Group is different from Chinese Medicine Low Dose Group significantly(P<0.01)、Darin-35 Group(p<0.01)and Chinese Medicine High Dose Group(P<0.05),indicating that metformin has the best effect on IR,chinese medicine of high dose takes the second place,Darin-35 has no significat effect.The TG,LDL,TC of the rats in the model blank group were significantly different from those of the normal control group(**P<0.01).TC of Chinese medicine groups and Diane group were higher than that of metformin group,the difference was significant(**P<0.01),shows that metformin can reduce TC better than the other drug intervention do.The level of TG and LDL in Chinese Medicine Group of each dose are significantly lower than that of the model control group,and the difference was significant(**P<0.01),indicating that the Chinese medicine has the effect of reducing TG and LDL.There was no significant difference between the TG and the metformin group in each dose groups,indicating that the two groups had the same effect.In the model group,miR-743b-3p、miR-743b-5p、miR-202-5p、miR-465-3p、miR-4635p、miR-881-3p、miR-463-3p、miR-881-3p、miR-463-3p、miR-741-3p、miR-3580-5p、miR-871-5p、miR-547-3p、miR-871-3p、miR-743a-3p、miR-743a-5p、miR-465-5p、miR471-5p、miR-878、miR-542-3p、miR-322-5p、miR-471-3p、miR-19a-3p...the expression of these miRNAs were down,miR-369-5p、miR-10a-5p、miR-187-3p、miR-187-5p、miR-3693p、miR-540-5p、miR-376b-3p、miR-136-5p、miR-10a-3p、miR-672-3p、miR-449a-5p、miR-449c-5p、miR-34c-5p、miR-92b-5p、miR-34b-5p、miR-34b-3p、miR-34c-3p...the expression of these miRNAs were up.The amount of expression of known Mature miRNA in each group:the expression of miR-135a is upregulated after modeling,the Metformin Group has the most obvious down-regulated expression,Chinese Medicine Group has down-regulated expression,while Darin-35 Group hasn’t.The expression of miR-145a(miR-145a-3p,miR-145a-5p)was up-regulated after the model,and the expression of metformin and Chinese medicine group was down regulated after the drug intervention.MiR-146a(miR-146a-5p,miR-146a-3p)upregulated after modeling.Darin-35 Group has down-regulated experssion,while metformin and Chinese Medicine Group still has a slightly up-regulated experssion.(4)In the clinical study of adolescent PCOS patients,the effect on lowering T level of Bushen Ditan Decoction was better than that of Metformin Group(P<0.05).Integrative Medicine Group improved the insulin resistance better than the Chinese medicine group(P<0.05).Bushen Ditan Decoction is superior to metformin in improving TCM syndromes and lowering testosterone level.(P<0.05),BuShen Ditan Decoction and Integrative Medicine Group have the same effect as metformin in reducing body mass index(BMI)and waist to hip ratio(WHR).Conclusion:The rat model of PCOS kidney deficiency and phlegm dampness lost normal estrous cycle,the ovary was polycystic change,androgen level was high,and IR and ovulation disorder appeared.Bushen Ditan Decoction can significantly improve the rat sex hormone,IR and lipid metabolism,the effect of lowering the level of T is better than that of Diane-35 and Metformin;Bushen Ditan Decoction could reduce the body weight of rats in PCOS model group;lowering T effect has been observed in each Chinese Medicine Group;Chinese Medicine Group and Diane-35 group have better effect on lowering T level than that of Metformin Group;Bushen Ditan Decoction can reduce TG and LDL levels;in improving IR,metformin has the best effect,Chinese Medicine High Dose Group takes the second place,Darine-35 has no significant effect.The differential expression profiles of miRNAs is found in the ovarian tissues of model groups and normal group,and there were different expressions between Metformin Group and Chinese Medicine Group after the drug intervention.Bushen Ditan Decoction can improve the syndrome of the patients with adolescent PCOS of IR and reduce androgen,and the effect is superior to metformin.In reducing body mass index(BMI)and waist hip ratio(WHR),Bushen Ditan Decoction has the similar effect with Metformin;in improving IR,metformin was better than Bushen Ditan Decoction.In general,Integrative Medicine Group has the best effect. |
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