| Objective:To explore the effects of Supplemented Banxia Xiexin Decoction on reproductive endocrine,glucose metabolism,inflammatory factors and intestinal flora in PCOS-IR model rats.Methods:A total of 78 female SD rats,aged 21 days,were randomly divided into blank control group(group Z,n=15)and model group(n=63).PCOS-IR rat model was established using letrozole combined with high fat diet.After successful modeling,the model group was randomly divided into model control group(group M),metformin hydrochloride group(group X),high-dose Chinese medicine group(group A),medium-dose chinese medicine group(group B),and low-dose chinese medicine group(group C),which were given corresponding medicine to each group for 2 weeks,while group M and group Z were given the same volume of normal saline.After the end day of the treatment,the glucose of rats was measured by glucose meter before abdominal anesthesia,and the feces of rats were collected by pressing rectum method,and the intestinal flora was detected by 16S rRNA technique.Serum estradiol,testosterone,luteinizing hormone,follicle-stimulating hormone,fasting insulin,lipopolysaccharide,tumor necrosis factor-a,interleukin-6 were determined by ELISA,and calculate LH/FSH ratio and HOMA-IR,and collected left ovarian tissue,using HE staining method to observe ovarian pathological changes.Results:1.General condition of rats:Compared with group Z,the weight of rats in group M increased significantly(P<0.001),and the ovarian organ coefficient decreased significantly(P<0.001).Compared with group M,each treatment group with the weight of rats decreased(P<0.05),and the ovarian organ coefficient increased significantly(P<0.05).No significant changes in ovarian weight in each group(P>0.05).2.Ovarian morphology:In group Z,there were multistage follicles and mature follicles with neat arrangement and multi-layer distribution of granulosa cells,with a large number of corpus luteum.In group M,no mature follicles were observed,only irregular and vacuolar dilated follicles,with fewer layers of granulosa cells and no corpus luteum.Compared with group M,the layers of granulosa cells and corpus luteum increased in each treatment group,and mature follicles were observed in group A.3.Reproductive endocrine:In terms of reproductive hormones,the level of T,FSH,LH,LH/FSH in group M increased significantly when compared with group Z(P<0.05),while the level of E2 was not significantly changed(P>0.05).Compared with group M,the level of T in group X decreased significantly(P<0.05),and the level of E2 increased significantly(P<0.01);the T level in group A had a decreasing trend(P>0.05),and in group B and group C,the level of E2 in Significantly increased,LH/FSH level decreased,which was statistically significant.4.Glucose metabolism:In terms of glucose metabolism,the levels of Glu and HOMA-IR in group M increased significantly when compared with group Z,the difference was statistically significant,while the level of FINS had a rising trend(P>0.05).Compared with group M,Glu and HOMA-IR in group X had a downward trend(P>0.05);in group A,HOMA-IR levels declined(P<0.05),and Glu and FINS levels had a downward trend(P>0.05);in group B,FINS and HOMA-IR have a downward trend(P>0.05).5.Inflammatory factors:Compared with group Z,the level of LPS in group M was significantly increased(P<0.001),and TNF-α had a rising trend(P>0.05).Compared with group M,the levels of LPS and TNF-α levels in group A decreased(P<0.05);TNF-α and IL-6 levels in group B decreased(P<0.05);all the inflammatory factors in group C have no Significant change(P>0.05).6.Results of intestinal flora:(1)Analysis of a diversity:Compared with group Z,the sobs index of group M increased(P<0.05),and the chao index,ace index and Shannon index showed an increasing trend(P>0.05),indicating that group M is rich in flora Increased degree and diversity.Compared with group M,the sobs index and ace index of each treatment group decreased significantly,especially in group A(P<0.001);the chao index and Shannon index of group X,A and C decreased significantly,and the chao index and Shannon index in group B showed a downward trend(P>0.05),which indicating that the abundance and diversity of microbial communities in each group decreased after treatment.(2)β diversity analysis shows that the distance between group Z and group M is the farthest,indicating that there are obvious differences in the composition of bacterial communities between the two groups;there is a significant gap between group M and other treatment groups,indicating that treatment group and group M have differences in the composition of the bacterial community,and the difference between group A and group M is the largest;the distance between the treatment groups is relatively close,indicating that there is a similarity in the community composition between the treatment groups.(3)Analysis of species composition:In terms of phylum level,the abundance of Verrucomicrobia in group M decreased(P<0.05),and the abundance of Bacteroidetes and Actinobacteria showed an upward trend(P>0.05).Compared with the M group,the abundance of Verrucomicrobia in each treatment group showed an upward trend(P>0.05),and the abundance of Bacteroidetes and Actinobacteria in group X decreased(P<0.05).The abundance of Bacteroidetes(Bacteroidetes)and Actinobacteria(Actinobacteria)in group A has a downward trend(P>0.05).In terms of genus level,compared with group Z,Akkermansia and Allobaculum in group M decreased(P<0.05),while Romboutsia,norank＿f＿Muribaculaceae,and Clostridium＿sensu＿stricto＿1 had an increasing trend(P>0.05).Compared with the M group,the Akkermansia genus in each treatment group has an increasing trend,and the Romboutsia has a decreasing trend(P>0.05);the Allobaculum genus in each treatment group has a different degree of increase,and group C has a statistical increase Significance(P<0.05),norank＿f＿Muribaculaceae has different degrees of reduction,among while group X is statistically significant(P<0.01);the Clostridium＿sensu＿stricto＿1 of each TCM treatment group is significantly decreased,and the difference is statistically significant significance.(4)Correlation analysis:the analysis of the main genera showed that Akkermansia was negatively correlated with weight,FINS,HOMA-IR,Glu and LPS(P>0.05),and positively correlated with LH/FSH(P<0.05).Conclusion:1.The PCOS-IR model rats have the imbalance of intestinal flora,which can led to the body’s inflammatory response;2.Supplemented Banxia Xiexin Decoction can improve the disorders of reproductive endocrine and glucose metabolism in PCOS-IR rat model,and can promote follicular development.Adjusting intestinal flora and improving inflammatory state is one of the ways to play its role. |
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