“秩边”透“水道”针刺对早发性卵巢功能不全大鼠死亡受体通路Fas/FADD/Caspase-8表达的影响

Effect of"Zhibian"(BL 54)-to-"Shuidao"(ST 28)needle insertion on the expression of Fas/FADD/Caspase-8 of death receptor pathway in rats with primary ovarian insufficiency

目的:基于死亡受体通路Fas/Fas相关死亡域蛋白(FADD)/半胱氨酸天冬氨酸蛋白酶-8(Caspase-8探讨“秩边”透“水道”针刺对早发性卵巢功能不全(POI)大鼠卵巢功能的影响及可能机制。方法:将48只雌性SD大鼠随机分为空白组、模型组、药物组和针刺组,每组12只。除空白组外,其余各组大鼠均采用腹腔注射环磷酰胺制备POI模型。针刺组大鼠于造模后采用“秩边”透“水道”针刺干预4周,每日1次,每次30 min;药物组予戊酸雌二醇片(0.09 mg·kg^(-1)·d^(-1))灌胃4周。比较各组大鼠一般情况和动情周期;采用ELISA法检测大鼠血清卵泡刺激素(FSH)、黄体生成素(LH)和雌二醇(E2)水平,HE染色法观察大鼠卵巢组织形态,免疫组化法和Western blot法检测大鼠卵巢组织Fas、FADD、Caspase-8蛋白表达。结果:造模后,除空白组外,其他各组大鼠毛发干焦、脱毛,精神萎靡,进食减少,小便增多,大便稀软不成形;干预后,针刺组、药物组大鼠大便逐渐成形。干预后,与空白组比较,模型组大鼠动情周期紊乱率升高(P<0.01);与模型组比较,针刺组、药物组大鼠动情周期紊乱率降低(P<0.01)。与空白组比较,模型组大鼠体质量和血清E2水平降低(P<0.01),血清FSH、LH水平和卵巢组织Fas、FADD、Caspase-8蛋白表达升高(P<0.01);与模型组比较,针刺组、药物组大鼠体质量和血清E2水平升高(P<0.01),血清FSH、LH水平和卵巢组织Fas、FADD、Caspase-8蛋白表达降低(P<0.01)。结论:“秩边”透“水道”针刺可有效改善POI大鼠卵巢功能,其机制可能为调节POI大鼠血清性激素水平,降低卵巢组织Fas、FADD、Caspase-8表达水平,减缓卵巢细胞凋亡。

Objective To explore the effect of"Zhibian"(BL 54)-to-"Shuidao"(ST 28)needle insertion on the ovarian function in the rats with primary ovarian insufficiency(POI)and the potential effect mechanism based on the Fas/FADD/Caspase-8 of death receptor pathway.Methods Forty-eight female SD rats were randomly divided into a blank group,a model group,a medication group and an acupuncture group,with 12 rats in each group.Except in the blank group,the rats in the other groups were intraperitoneally injected with cyclophosphamide to establish the POI model.In the acupuncture group,after successful modeling,the intervention was given with"Zhibian"(BL 54)-to-"Shuidao"(ST 28)needle insertion,once daily,30 min in each intervention;and the duration of intervention was 4 weeks.In the medication group,estradiol valerate tablets were administered intragastrically,0.09 mg·kg^(-1)·d-1,for 4 weeks.The general situation and the estrous cycle of the rats were compared among groups.Using ELISA,the levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E2)in the serum were detected.HE staining was adopted to observe the morphological changes of ovarian tissue of rats.The protein expression of Fas,FADD and Caspase-8 in ovarian tissue was detected with immunohistochemistry and Western blot.Results After modeling,except the rats of the blank group,the rats of the other groups had dry fur,lost hair,low spirits,reduced food intake,increased urination and loose stool.After intervention,the stool became regular gradually in the acupuncture group and the medication group.The percentage of estrous cycle disturbance was increased in the rats of the model group when compared with the blank group(P<0.01);in comparison with the model group,the percentages of estrous cycle disturbance were reduced in the acupuncture group and the medication group after intervention(P<0.01).When compared with the blank group,the body mass and E2 content in the serum were lower(P<0.01),the levels of FSH and LH in the serum and the protein expression levels of Fas,FADD and Caspase-8 were increased(P<0.01)in the model group.Compared with the model group,the body mass and E2 contents in the serum were higher(P<0.01),the levels of FSH and LH in the serum and the protein expression levels of Fas,FADD and Caspase-8 were reduced(P<0.01)in the acupuncture group and the medication group.Conclusion"Zhibian"(BL 54)-to-"Shuidao"(ST 28)needle insertion can effectively improve the ovarian function of POI rats,and its effect mechanism may be related to regulating the serum sex hormone levels,reducing the expression of Fas,FADD and Caspase-8 in ovarian tissue and retarding apoptosis of ovarian cells.