针刺对多囊卵巢综合征胰岛素抵抗大鼠生殖内分泌及磷酸化蛋白激酶B蛋白表达的影响

Effect of Acupuncture on Reproductive Endocrine and p-Akt Protein Expression in Polycystic Ovary Syndrome Rats with Insulin Resistance

目的:研究针刺对多囊卵巢综合征胰岛素抵抗(PCOS-IR)大鼠生殖内分泌激素及磷酸化蛋白激酶B(p-Akt)蛋白表达的影响,探讨针刺改善PCOS-IR的作用机制。方法:雌性Wistar大鼠随机分为空白对照组、模型组、二甲双胍组、针刺组,每组8只。采用背部皮下注射脱氢表雄酮建立PCOS-IR模型。针刺组针刺双侧"肾俞""胰俞""子宫""三阴交""丰隆"及"关元"穴,提插捻转5min;二甲双胍组给予二甲双胍溶液(100mg/kg)灌胃,两组均每日治疗1次,连续治疗28d。采用HE染色观察卵巢组织变化;ELISA法检测血清卵泡刺激素(FSH)、雌二醇(E2)、黄体生成素(LH)和睾酮(T)水平及空腹胰岛素(FINS)水平,并计算胰岛素抵抗指数(HOMA-IR);Western blot法检测卵巢组织Akt和p-Akt蛋白表达情况。结果:与正常组比较,模型组卵巢呈多囊样改变,闭锁卵泡增多,未见黄体分布,且模型组血清FSH、E2明显降低(P<0.01),血清LH和T水平明显升高(P<0.01),FINS水平和HOMA-IR值升高(P<0.01),卵巢组织Akt和p-Akt蛋白表达明显降低(P<0.01,P<0.05)。与模型组比较,针刺组和二甲双胍组可见不同发育阶段的卵泡及少量黄体,血清FSH、E2水平明显升高(P<0.01),血清LH、T水平明显降低(P<0.05,P<0.01),FINS水平及HOMA-IR值降低(P<0.01),卵巢组织Akt和p-Akt蛋白表达明显升高(P<0.05,P<0.01)。针刺组和二甲双胍组各指标的差异无统计学意义(P>0.05)。结论:根据中医生殖轴选取的穴位针刺可以改善PCOS-IR大鼠生殖内分泌水平以及IR情况,这可能与针刺对卵巢组织p-Akt蛋白表达的调控有关。

Objective To observe the effect of manual acupuncture intervention on the reproductive endocrine and p-Akt protein expression in polycystic ovary syndrome with insulin resistance（PCOS-IR）rats,so as to reveal its mechanism in relieving PCOS-IR.Methods Female Wistar rats were randomly divided into control,model,medication（Metformin）and acupuncture groups（n=8 in each）.The PCOS-IR model was established by subcutaneous injection of Dehydroepiandrosterone（DHEA,6 mg/100 g）,once daily for 20 days.Rats of the medication group were treated by gavage of Metformin solution（100 mg/kg）,once daily for 28 days.For rats of the acupuncture group,manual acupuncture stimulation（lifting-thrusting and rotating of needles）was applied to the bilateral＂Shenshu＂（BL 23）,＂Yishu＂（EX-B 3）,＂Zigong＂（EX-CA 1）,＂Sanyinjiao＂（SP 6）,＂Fenglong＂（ST 40）and＂Guanyuan＂（CV 4）for 5 min,once a day for 28 days.H.E.staining was used to observe the histopathologic changes of the ovary tissue.Serum levels of follicle-stimulating hormone（FSH）,estradiol（E2）,luteinizing hormone（LH）,testosterone（T）and fasting insulin（FINS）were measured by ELISA.Fasting plasma glucose（FPG）was measured by blood glucose meter,and homeostasis model assessment-insulin resistance index（HOMA-IR）was calculated.Western blot was used to detect the expression of Akt and p-Akt proteins in ovarian tissues.Results Following modeling,the ovaries showed polycystic changes and an increase of atresia follicles with a larger diameter and no corpus luteum in the model group.After the intervention,there were various types of follicles and a small amount of corpus luteum at different stages of development in both acupuncture and medication groups.In comparison with the control group,serum FSH and E2 levels as well as ovarian Akt and p-Akt protein expression were considerably decreased（P〈0.05,P〈0.01）,and serum LH,T and FINS contents and HOMA-IR were significantly increased in the model group（P〈0.01）.Following the intervention,the decreased serum FSH and E2 levels as well as ovarian Akt and p-Akt protein expression,and the increased serum LH,T and FINS contents and HOMA-IR were remarkably reversed in both acupuncture and medication groups（P〈0.05,P〈0.01）,without significant differences between the two groups in the levels of those serum and ovary indexes（P〈0.05）.Conclusion Acupuncture intervention can regulate the levels of reproductive endocrine hormones and reduce insulin resistance in PCOS-IR rats,which may be related to its effect in up-regulating the expression of p-Akt protein in the ovaries.