摘要
目的基于Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路探讨小檗碱联合槲皮素对多囊卵巢综合征(PCOS)大鼠的影响。方法将30只SD雌性大鼠根据体重分层随机分为空白对照组、模型对照组、小檗碱组、槲皮素组及小檗碱+槲皮素组,每组6只。在大鼠皮下每天注射脱氢表雄酮,构建PCOS模型。药物连续干预21 d后,称重各组大鼠体重和卵巢质量,苏木精-伊红(HE)染色法对大鼠卵巢组织进行病理形态学观察,ELISA测定血清中黄体生成素(LH)、促卵泡生成素(FSH)、睾酮(T)、白细胞介素-6(IL-6)、肿瘤细胞因子-α(TNF-α)水平,蛋白免疫印迹法(Western blot)测定TLR4、NF-κB蛋白相对表达水平。结果与空白对照组相比,模型对照组卵巢的表现为多囊样,闭锁卵泡增多,体重和卵巢质量均增加(P<0.01),LH、T、IL-6和TNF-α水平升高(P<0.01),FSH水平降低(P<0.01),TLR4和磷酸化NF-κBp65(p-NF-κBp65)蛋白相对表达水平升高(P<0.01)。与模型对照组相比,小檗碱组、槲皮素组和小檗碱+槲皮素组卵巢囊性扩张降低,闭锁卵泡减少,局部卵巢形态与空白对照相似;与模型对照组相比,小檗碱组体重降低(P<0.05),小檗碱组和槲皮素组中LH、T、IL-6、TNF-α水平降低(P<0.01),TLR4和p-NF-κBp65蛋白相对表达水平降低(P<0.01),卵巢质量也降低(P<0.01);但小檗碱组和槲皮素组FSH水平及体重比较,差异无统计学意义(P>0.05)。小檗碱+槲皮素组体重和卵巢质量均降低(P<0.01),FSH水平升高(P<0.01),LH、T、IL-6和TNF-α水平均降低(P<0.01),TLR4和p-NF-κBp65蛋白相对表达水平降低(P<0.01)。结论小檗碱联合槲皮素可能通过抑制TLR4/NF-κB信号通路,调节FSH、LH、T分泌,减少IL-6、TNF-α炎症因子的释放,改善卵巢功能。
Objective To investigate the effects of berberine combined with quercetin on the rats with polycystic ovary syndrome(PCOS)based on toll-like recepter 4(TLR4)/nuclear-factor-κB(NF-κB)signaling pathway.Methods Thirty SD female rats were stratified by weight and then randomly divided into the blank control group,model control group,berberine group,quercetin group and berberine+quercetin group,6 rats in each group.The rats conducted daily subcutaneous injection of dehydroepiandrosterone(DHEA)for constructing the PCOS model.After consecutive medication intervention for 21 d,the body weight and ovary mass of the rats in each group were weighed,and the hematoxylin-eosin(HE)staining was used for conducting the histological morphological observation on ovarian tissues.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum luteinizing hormone(LH),follicle stimulating hormone(FSH),testosterone(T),interleukin-6(IL-6)and tumor cytokine alpha(TNF-α).The Western blot was used to determine the relative levels of TLR4 and NF-κB protein.Results Compared with the blank control group,the ovaries in the model control group were manifested by polycystoid,atretic follicles increase,body weight and ovarian mass increase(P<0.01),the increase of LH,T,IL-6 and TNF-αlevels(P<0.01),decrease of FSH level(P<0.01)and increase of TLR4 and p-NF-κBp65 protein expression levels(P<0.01).Compared with the model control group,the ovarian cystic dilatation in the berberine group,quercetin group and the berberine+quercetin group was decreased,the atretic follicles were decreased,the local ovarian morphology was similar to that of the blank control group.Compared with the model control group,the body weight in the berberine group was decreased(P<0.05),the levels of LH,T,IL-6 and TNF-αin the berberine group and quercetin group were decreased(P<0.01),the levels of TLR4 and p-NF-κBp65 protein expression were decreased(P<0.01)and the ovarian mass was reduced(P<0.01).But the FSH and the body weight level had no statistical difference between the berberine group and quercetin group(P>0.05).The body mass and ovarian mass in the berberine+quercetin group were decreased(P<0.01),the FSH level was increased(P<0.01),the LH,T,IL-6 and TNF-αlevels were decreased(P<0.01),and the TLR4 and p-NF-κBp65 protein expression levels were decreased(P<0.01).Conclusion Berberine combined with quercetin may regulate the FSH,LH and T hormone secretion,decrease the release of IL-6 and TNF-αinflammatory factors and improve the ovarian function possibly by inhibiting the TLR4/NF-κB signaling pathway.
作者
杨菡
林思怡
谭嘉哲
马泽茜
张晓君
朱乐玫
YANG Han;LIN Siyi;TAN Jiazhe;MA Zeqian;ZHANG Xiaojun;ZHU Lemei(School of Clinical Sciences,Changsha Medical College,Changsha,Hunan 410219,China;School of Public Health,Changsha Medical College,Changsha,Hunan 410219,China;College of Stomatology,Changsha Medical College,Changsha,Hunan 410219,China;College of Traditional Chinese Medicine,Changsha Medical College,Changsha,Hunan 410219,China)
出处
《重庆医学》
CAS
2024年第23期3540-3545,3551,共7页
Chongqing Medical Journal
基金
国家级大学生创新训练计划项目(教高司函〔2023〕8号-S202310823017
教高司函〔2021〕13号-S202110823026)。
