电针“公孙”调节Keap1/Nrf2/HO-1通路对早发性卵巢功能不全大鼠氧化应激损伤的影响  

Electroacupuncture of“Gongsun”(SP4)alleviates oxidative stress injury and promotes normal follicle development by regulating keap1/Nrf2/HO-1 signaling in rats with premature ovarian insufficiency

作　　者：黄思丹 张怡 师旭亮 张丹妮 李欣悦 孙青松 佘延芬 HUANG Si-dan;ZHANG Yi;SHI Xu-liang;ZHANG Dan-ni;LI Xin-yue;SUN Qing-song;SHE Yan-fen(Hebei International Joint Research Center for Dominant Diseases in Traditional Chinese Medicine Acupuncture and Moxibustion,Shijiazhuang 050091,China;College of Acupuncture-moxibustion and Tuina,Hebei University of Chinese Medicine,Shijiazhuang 050020;Dou Han-qing Academic Thought Research Institute of Hebei University of Chinese Medicine,Shijiazhuang 050020)

机构地区：[1]河北省中医针灸优势病症国际联合研究中心,石家庄050091 [2]河北中医药大学针灸推拿学院,石家庄050020 [3]河北中医药大学窦汉卿学术思想研究院,石家庄050020

出　　处：《针刺研究》2025年第1期41-49,共9页Acupuncture Research

基　　金：河北省自然科学基金项目(No.H2022423371);河北省中医药管理局项目(No.2022093);国家自然科学基金项目(No.81973755);河北中医药大学“大学生创新创业训练计划”项目(No.202314432023)。

摘　　要：目的:观察电针“公孙”对早发性卵巢功能不全(POI)大鼠Kelch样环氧氯丙烷相关蛋白因子1(Keap1)/核因子E_(2)相关因子2(Nrf2)/血红素氧合酶-1(HO-1)通路的影响,探讨电针“公孙”治疗POI的作用机制。方法:将32只雌性大鼠随机分为空白组、模型组、电针组及西药组,每组8只。采用腹腔注射环磷酰胺制备POI大鼠模型。电针组大鼠给予电针干预双侧“公孙”20 min,每日1次,持续14 d;西药组予戊酸雌二醇溶液灌胃,每日1次,持续14 d。实验过程中记录各组大鼠体质量变化并观察一般情况。干预结束后,ELISA检测大鼠血清促卵泡激素(FSH)、雌二醇(E_(2))、抗米勒管激素(AMH)、丙二醛(MDA)含量及过氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活力;HE染色法观察卵巢组织形态变化;免疫组织化学法和Western blot检测卵巢组织Keap1、Nrf2和HO-1蛋白表达;PCR法检测卵巢组织Keap1、Nrf2和HO-1mRNA表达。结果:与空白组比较,造模后各组大鼠精神状态欠佳,可见躯体消瘦,毛发干枯、无光泽,偶见黄色稀软便、黑便、眼周鼻尖有带血分泌物,动情周期紊乱。干预后,电针组与西药组大鼠体质量上升,大便呈棕褐色、卵圆形,未见带血分泌物。与空白组比较,模型组大鼠血清FSH、MDA水平升高(P<0.01),E_(2)、AMH水平及SOD、GSH-Px活力降低(P<0.01),各级生长卵泡数量减少,闭锁卵泡增加,卵巢组织中Keap1阳性表达增加(P<0.01),Nrf2、HO-1阳性表达减少(P<0.01),卵巢组织中Keap1蛋白表达增加(P<0.01),Nrf2、HO-1蛋白表达减少(P<0.01),卵巢组织中Keap1 mRNA表达增加(P<0.01),Nrf2、HO-1 mRNA表达减少(P<0.01);与模型组比较,电针组、西药组大鼠血清FSH、MDA水平降低(P<0.01),E_(2)、AMH水平及SOD、GSH-Px活力升高(P<0.01),各级生长卵泡数量增加,闭锁卵泡减少,卵巢组织中Keap1阳性表达减少(P<0.01),Nrf2、HO-1阳性表达增加(P<0.05,P<0.01),卵巢组织中Keap1蛋白表达减少(PObjective Based on the theory of"Gongsun(SP4)connects the Thoroughfare Vessel"of the"eight confluence acupoints",the effect of electroacupuncture(EA)at SP4 on Kelch-like epichlorohydrin associated protein factor 1(Keap1)/nuclear factor-erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway was observed in rats with premature ovarian insufficiency(POl)for exploring its underlying mechanisms in treating POl.Methods Thirty-two female SD rats were randomly divided into 4 groups:blank,model,EA and medication,with 8 rats in each group.The POl model was established by intraperitoneal injection of cyclophosphamide.EA(2 Hz,a strength inducing slight limb tremor)was applied to bilateral SP4 for 20 min,once a day for a total of 14 d.Rats of the medication group were treated by gavage of solution of estradiol valerate tablets(0.09 mg/kg)once a day for 14 d.During the experiment,the changes of body mass in each group were recorded and the general conditions were observed.After the interventions,the contents of follicle-stimulating hormone(FSH),estradiol(E_(2)),anti-mullerian hormone(AMH),malondialdehyde(MDA)and theactivity ofsuperoxidedismutase(SOD),glutathione peroxidase(GSH-Px)were detected by ELISA.The morphological changes of ovarian tissue were observed under light microscope after H.E.staining.The protein and mRNA expression levels of Keap1,Nrf2 and HO-1 in the ovarian were detected by immunohistochemistry,Western blot,and PcR,respectively.Results After modeling,the model rats had poor mental state,reduced activity,irritability,emaciated body,dry and dull hair,decreased food intake,and occasionally yellow soft stools,black stools,and bloody secretions around the eyes and the tip of the nose.After the interventions,the mental state of rats in the EA and medication groups was recovered,and had brown oval feces,with no bloody secretion.Compared with the blank group,the body mass,levels of serum E_(2),AMH,SOD,GSH-Px,immunoactivity of Nrf2 and HO-1,and expression levels of Nrf2 and HO-1 proteins and mRNAs

关 键 词：早发性卵巢功能不全 电针 公孙 氧化应激 Kelch样环氧氯丙烷相关蛋白因子1/核因子E2相关因子2/血红素氧合酶-1信号通路 

分 类 号：R245.97[医药卫生—针灸推拿学]