电针对PCOS合并IR大鼠下丘脑瘦素/Kisspeptin信号的影响

目的:通过电针对多囊卵巢综合征(PCOS)合并胰岛素抵抗(IR)大鼠生殖内分泌与代谢功能的同时调节,探讨其与影响下丘脑瘦素(LEP)/Kisspeptin(KP)信号传导的关联性。方法:3周龄SD雌性大鼠以硫酸普拉睾酮钠皮下注射联合高脂饲料喂养21 d建立PCOS合并IR模型,建模后随机分为模型组、西药组、电针组,继续高脂饲料喂养。另设空白组,每组8只。电针组大鼠予三阴交、关元、中脘、足(后)三里穴位电针治疗,西药组大鼠予二甲双胍溶液灌胃,连续治疗28 d。检测血清T、LH、FSH、LEP、FINS和FPG水平变化,计算HOMA-IR;HE染色观察卵巢形态学变化;实时PCR法检测下丘脑LEP mRNA、Kiss-1 mRNA和GnRH mRNA表达水平;蛋白质印迹法检测下丘脑LEP和KP蛋白表达水平。结果:与空白组比较,模型组大鼠性激素紊乱、糖脂代谢异常(P<0.01),下丘脑LEP mRNA及蛋白表达降低且Kiss-1 mRNA、GnRH mRNA及KP蛋白表达升高(P<0.01)。与模型组比较,电针组大鼠血清LH、LEP水平及HOMA-IR降低(P<0.01或P<0.05),下丘脑LEP mRNA及蛋白表达升高(P<0.01或P<0.05),GnRH mRNA表达降低(P<0.05);西药组大鼠血清T、LH、LEP、FINS水平及HOMA-IR降低(P<0.05或P<0.01),下丘脑LEP mRNA表达升高(P<0.01)。西药组和电针组大鼠卵巢结构均较模型组改善,电针组大鼠血清T、LH、FSH、LEP、FINS、FPG水平,HOMA-IR,下丘脑LEP mRNA、Kiss-1 mRNA及GnRH mRNA表达,以及LEP及KP蛋白表达与西药组比较,差异均无统计学意义(P>0.05)。结论:电针干预PCOS合并IR大鼠,可影响下丘脑LEP/KP信号传导,可能是电针同时调节糖脂代谢与生殖内分泌治疗PCOS的机制。

Effect of Electroacupuncture on Leptin/Kisspeptin Signal of Hypothalamus in PCOS and IR Rats

Objective: To investigate the effect of electroacupuncture on the reproductive endocrine and metabolic functions of rats with polycystic ovary syndrome(PCOS) combined with insulin resistance(IR), and to explore the relationship between leptin(LEP)/Lisspeptin(KP) signal transduction in hypothalamus. Methods: 3-week-old SD female were fed with prasterone sulfate sodium subcutaneously combined with high-fat diet for 21 d to establish the PCOS combined IR model. After modeling, rats were randomly divided into model group, western medicine group and electroacupuncture group, and continued high-fat diet feed. There is also a blank group with 8 rats.Rats in the electroacupuncture group were treated with electroacupuncture at "Sanyinjiao"(SP6), "Guanyuan"(BL26), "Zhongwan"(RN12), and "Zu(hou) Sanli"(ST36) points. Rats in the western medicine group were treated with metformin solution by gavage for 28 consecutive days. The changes of serum T, LH, FSH, LEP, FINS and FPG levels were detected, and HOMA-IR was calculated. The morphological changes of ovary were observed by HE staining. The expression levels of LEP mRNA, Ki SS-1 mRNA and GnRH mRNA in hypothalamus were detected by real-time PCR. The protein expression levels of LEP and KP in hypothalamus were detected by Western blotting. Results: Compared with the blank group, the sex hormone disorder, abnormal glucose and lipid metabolism were observed in the model group(P<0.01), the expression of LEP mRNA and protein in hypothalamus was decreased, and the expression of Kiss-1 mRNA, Gn RH mRNA and KP protein was increased(P<0.01). Compared with the model group, the levels of serum LH, LEP and HOMA-IR in electroacupuncture group were decreased(P<0.01 or P<0.05), the expression of LEP mRNA and protein in hypothalamus was increased(P <0.01 or P <0.05), and the expression of GnRH mRNA was decreased(P<0.05). The levels of serum T, LH, LEP, FINS and HOMA-IR were decreased in western medicine group(P <0.05 or P <0.01), and LEP mRNA expression in hypothalamus was increased(P <0.01). The ovarian structure of rats in western medicine group and electroacupuncture group was better than that in model group. There were no significant differences in serum T, LH,FSH, LEP, FINS, FPG levels, HOMA-IR, LEP mRNA, Kiss-1 mRNA and Gn RH mRNA expression in hypothalamus,as well as LEP and KP protein expression between electroacupuncture group and western medicine group(P >0.05). Conclusion: Electroacupuncture intervention in PCOS combined with IR rats can affect LEP/KP signal transduction in the hypothalamus, which may be the mechanism of electroacupuncture to simultaneously regulate glucose and lipid metabolism and reproductive endocrine therapy for PCOS.