朱琏抑制Ⅱ型针法对多囊卵巢综合征伴胰岛素抵抗大鼠IRS-1、IRS-2蛋白的影响

Effect of Zhu Lian's Type Ⅱ Inhibition Needling Technique on IRS-1 and IRS-2 Protein Expression on Polycystic Ovary Syndrome Combined with Insulin Resistance Model

  • 摘要: 目的  探讨朱琏抑制Ⅱ型针法改善多囊卵巢综合征(PCOS)伴胰岛素抵抗(IR)的作用和机制。方法  将40只雌性SD大鼠随机分为空白组、模型组、针刺组、西药组,每组10只,采用来曲唑联合高脂饲料喂养建立PCOS-IR模型。针刺组予朱琏抑制Ⅱ型针法针刺关元及双侧归来、足三里、三阴交、丰隆穴,留针30 min;西药组给予炔雌醇环丙孕酮0.18 mg/(kg·d)、二甲双胍135 mg/(kg·d)灌胃。每组均每日治疗1次,连续治疗28 d。经断尾采血法测定空腹血糖(FBG),ELISA法检测血清空腹胰岛素(FINS)水平,并计算胰岛素抵抗指数(HOMA-IR);免疫组化法检测卵巢组织胰岛素受体底物-1(IRS-1)和胰岛素受体底物-2(IRS-2)蛋白定位表达;Western blot法检测卵巢组织IRS-1和IRS-2蛋白相对表达量。结果  免疫组化染色结果显示,IRS-1、IRS-2蛋白在卵巢组织间质细胞、颗粒细胞、初级卵母细胞中均有表达。与空白组比较,模型组血清FINS水平和HOMA-IR值显著升高(P < 0.05),卵巢组织IRS-1和IRS-2表达明显降低(P < 0.05)。与模型组比较,针刺组和西药组血清FINS水平及HOMA-IR值显著降低(P < 0.05),卵巢组织IRS-1和IRS-2表达明显升高(P < 0.05)。针刺组和西药组各指标间比较差异均无统计学意义。结论  朱琏抑制Ⅱ型针法针刺治疗可以改善PCOS大鼠IR情况,这可能与良性调控卵巢组织IRS-1和IRS-2蛋白表达有关。

     

    Abstract: OBJECTIVE  To explore the effect and mechanism of Zhu Lian's type Ⅱ inhibition needling technique on regulating polycystic ovary syndrome with insulin resistance (PCOS-IR) in rats.METHODS  40 female SD rats were randomly divided into normal group, model group, acupuncture group and medication group, with 10 rats in each group. The acupuncture group were given "Guanyuan" (CV4) and bilateral "guilai" (ST29), "Zusanli" (ST36), "Sanyinjiao" (SP6), and "Fenglong" (ST40) points by Zhu Lian's type Ⅱ inhibition needling technique for 30 min, once a day for 28 d. The medication group was given ethinylestradiol cyproterone of 0.18 mg/(kg·d) and metformin of 135 mg/(kg·d) by gavage, once a day for 28 d. HE staining was used to observe the changes of ovarian tissue. Fasting blood glucose (FBG) was measured by tail cutting method. Fasting insulin (FINS) was measured by ELISA, and insulin resistance index (HOMA-IR) was calculated. The protein expressions of IRS-1 and IRS-2 in ovarian tissue were detected by Western blot.RESULTS  Immunohistochemistry result showed that IRS-1 and IRS-2 proteins were expressed in ovarian interstitial cells, granulosa cells and primary oocytes. Compared with the control group, the ovaries in the model group showed polycystic changes, with more preantral follicles, and the number of granulosa cells in the model group was less than that in the control group, and FINS content and HOMA-IR in the model group significantly increased (P < 0.05), and the protein expressions of IRS-1 and IRS-2 in ovarian tissue significantly decreased (P < 0.05). Compared with the model group, the preantral follicles and antral follicles were observed in the acupuncture group and medication (metformin) groups, the number of preantral follicles was less than that of the model group, the number of granulosa cell layers increased, the morphology was more complete, and the arrangement was orderly; FINS content and HOMA-IR significantly decreased (P < 0.05), and the expressions of IRS-1 and IRS-2 protein in ovarian tissue significantly increased (P < 0.05). There was no significant difference between acupuncture group and western medicine group.CONCLUSION  Zhu Lian's type Ⅱ inhibition needling technique can improve IR in PCOS rats, which may be related to the benign regulation of IRS-1 and IRS-2 protein expressions in ovarian tissue.

     

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