鹅去氧胆酸对多囊卵巢综合征大鼠内分泌状态及胰岛功能的影响

Effects of chenodeoxycholic acid on endocrine status and islet function in rats with polycystic ovary syndrome

目的探索鹅去氧胆酸(CDCA)对多囊卵巢综合征(PCOS)大鼠内分泌功能及胰岛功能的影响,为进一步完善PCOS的临床治疗提供基础。方法选取30只SPF级SD大鼠为研究对象,分为对照组(5只)和模型组(25只),模型组大鼠使用来曲唑—羧甲基纤维素钠混悬液灌胃构建大鼠PCOS模型,造模成功后将模型组大鼠分为PCOS组和PCOS+CDCA组。PCOS组继续予以来曲唑—羧甲基纤维素钠混悬液灌胃,PCOS+CDCA组在PCOS组基础上予以0.3 mL 0.25%鹅去氧胆酸钠灌胃。并在饲养第21天及饲养第42天,采用HE染色观察模型组大鼠的卵巢形态;获取大鼠的尾静脉血,检测其血清睾丸素水平、空腹血糖、空腹胰岛素及炎症因子水平,计算胰岛素抵抗指数的稳态模型评估(HOMA-IR)和β细胞功能的稳态模型评估(HOMA-β)指数;收集模型组大鼠的粪便分析肠道菌群中大肠埃希菌、变形杆菌、链球菌及革兰阳性杆菌的占比。采用Spearman相关性分析评估模型组大鼠肠道菌群与其卵巢囊性卵泡数量、血清睾丸素、HOMA-IR、HOMA-β、肿瘤坏死因子-α(TNF-α)及白介素-6(IL-6)的相关性。结果模型组大鼠的囊性卵泡数量、血清睾丸素、HOMA-IR水平均显著高于对照组,HOMA-β水平显著低于对照组(均P<0.05),血清炎症因子水平显著高于对照组(均P<0.05)。PCOS组大鼠的血清睾丸素、HOMA-IR及炎症因子水平显著高于PCOS+CDCA组大鼠,而HOMA-β水平显著低于PCOS+CDCA组大鼠(均P<0.05)。PCOS组大鼠肠道大肠埃希菌比例呈显著上升趋势,而变形杆菌、链球菌及革兰阳性杆菌比例呈显著下降趋势(均P<0.05)。在CDCA的作用下,PCOS+CDCA组大鼠肠道大肠埃希菌比例升高趋势显著减缓,变形杆菌、链球菌及革兰阳性杆菌比例下降趋势显著减缓(均P<0.05);干预后PCOS+CDCA组大鼠的肠道大肠埃希菌占比显著低于PCOS组大鼠,而变形杆菌、链球菌及革兰阳性杆菌比例均显著高于PCOS组大鼠(均P<0.05)。PCOS组大鼠中,大肠埃希菌占比与囊性卵泡数量、血清睾丸素、HOMA-IR、TNF-α、IL-6均呈正相关,HOMA-β呈负相关(均P<0.05);变形杆菌、链球菌及革兰阳性杆菌的占比与囊性卵泡数量、血清睾丸素、HOMA-IR、TNF-α、IL-6均呈负相关,与HOMA-β呈正相关(均P<0.05)。在PCOS+CDCA组大鼠中,大肠埃希菌占比与囊性卵泡数量、TNF-α均呈正相关(均P<0.05),变形杆菌占比与血清睾丸素呈负相关(P<0.05)。结论CDCA可以在一定程度上缓解PCOS大鼠的内分泌紊乱、胰岛素抵抗及机体炎症水平,而这些疾病状态的改变与肠道菌群的调节具有显著的相关性。

Objective To explore the effects of chenodeoxycholic acid(CDCA)on endocrine function and islet function in polycystic ovary syndrome(PCOS)rats.Methods Thirty SPF SD rats were selected as the subjects and divided into control group and model group.The model group was given suspension of Letrozole and Carboxymethyl cellulose sodium(CMC-Na)by gavage to construct rat PCOS models.After successful modeling,the model group was divided into PCOS group and PCOS+CDCA group;both groups were given suspension of Letrozole and CMC-Na by gavage,and the PCOS+CDCA group was additionally given 0.3 mL 0.25%CDCA sodium.On the 21st and 42nd days of feeding,HE staining was used to observe the ovary morphology of rats in the model group.The tail vein blood of rats was collected for detection of the levels of serum testosterone,fasting blood glucose,fasting insulin and inflammatory factors.Homeostasis model assessment of insulin resistance index(HOMA-IR)and homeostasis model assessment ofβcell function(HOMA-β)were calculated.The feces of the model rats were collected for analysis of the proportion of Escherichia coli,Proteus,Streptococcus and Gram-positive bacilli in intestinal flora.Spearman correlation analysis was used to evaluate the correlation of intestinal flora with the number of ovarian cystic follicles,serum testosterone,HOMA-IR,HOMA-β,tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the model group.Results The number of cystic follicles and the levels of serum testosterone and HOMA-IR in model group were significantly higher,while the level of HOMA-βwas significantly lower than those in control group respectively;the levels of serum inflammatory factors in model group were significantly higher than those in control group(all P<0.05).The levels of serum testosterone,HOMA-IR and inflammatory factors in PCOS group were significantly higher,while the level of HOMA-βwas significantly lower than those in PCOS+CDCA group respectively(all P<0.05).The proportion of Escherichia coli in PCOS group significantly increased,while those of Proteus,Streptococcus and Gram-positive bacillus significantly decreased(all P<0.05).Under the action of CDCA,the proportion of Escherichia coli in PCOS+CDCA group significantly decreased(P<0.05).After intervention,the proportion of Escherichia coli in PCOS+CDCA group was significantly,while those of Proteus,Streptococcus and Gram-positive bacillus were significantly higher than those in PCOS group respectively(all P<0.05).In PCOS group,the proportion of Escherichia coli was positively correlated with the number of cystic follicles,serum testosterone,HOMA-IR,TNF-αand IL6,and negatively correlated with HOMA-β(all P<0.05).The proportion of Proteus,Streptococcus and Gram-positive bacilli were negatively correlated with the number of follicles,serum testosterone,HOMA-IR,TNF-αand IL-6,and positively correlated with HOMA-β(all P<0.05).In the PCOS+CDCA group,the proportion of Escherichia coli was positively correlated with the number of cystic follicles and TNF-α,while that of Proteus was negatively correlated with serum testosterone(P<0.05).Conclusion CDCA can relieve the internal dysfunction,insulin resistance and inflammation in PCOS rats to a certain extent,and the changes of these disease status are obviously related to the regulation of intestinal flora.