电针对卵巢早衰小鼠铁死亡的影响

Effect of electroacupuncture on ferroptosis in mice with premature ovarian failure

目的观察电针治疗卵巢早衰(premature ovarian failure,POF)的效果并探讨铁死亡在其中发挥的作用。方法将36只C57BL/6雌性小鼠随机分成对照组、模型组和电针组,每组12只,利用高脂高糖法造模后,以关元、双侧足三里和三阴交为针刺穴位进行电针干预。采用HE染色法观察小鼠卵泡的发育情况;酶联免疫吸附法检测小鼠血清卵泡生成素(follicle-stimulating hormone,FSH)和雌二醇(estradiol,E_(2))水平;利用分光光度法检测Fe^(2+)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)浓度;RT-q PCR法检测铁蛋白(ferritin)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、B淋巴细胞瘤2(B-cell lymphoma 2,BCL2)、BCL2相关X蛋白(BCL2-associated X,BAX)mRNA表达水平;Western blot法检测Ferritin、P53、GPX4、BCL2蛋白表达。结果电针治疗后成熟卵泡出现,FSH降低(P<0.01)、E_(2)升高(P<0.01);POF小鼠卵巢内氧化应激水平和Fe^(2+)浓度升高(P<0.01),而电针后SOD升高(P<0.01),MDA和Fe^(2+)降低(P<0.01);电针治疗后Ferritin和GPX4m RNA和蛋白表达显著升高(P<0.01),且电针可以促进BCL2的表达(P<0.01),抑制BAX和P53mRNA和蛋白的表达(P<0.01)。结论电针对POF具有良好的治疗效果,其机制可能通过靶向Xc-的GPX4抑制铁死亡和凋亡实现的。

Objective To observe the efficacy of electroacupuncture in treating premature ovarian failure(POF)and discuss the role of ferroptosis.Method Thirty-six female C57BL/6 mice were randomized into a control group,a model group,and an electroacupuncture group,with 12 mice in each group.Electroacupuncture was performed at Guanyuan(CV4)and bilateral Zusanli(ST36)and Sanyinjiao(SP6)after modeling with the high-lipid,high-glucose method.Hematoxylin-eosin(HE)staining was adopted to observe the growth of follicles in the mice;the enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum follicle-stimulating hormone(FSH)and estradiol(E_(2));the concentrations of Fe^(2+),superoxide dismutase(SOD),and malondialdehyde(MDA)were determined using spectrophotometry;the m RNA expression levels of ferritin,glutathione peroxidase 4(GPX4),B-cell lymphoma 2(BCL2),and BCL2-associated X(BAX)were examined using the real-time quantitative polymerase chain reaction(RT-qPCR)method;Western blot was adopted to detect the protein expression of ferritin,P53,GPX4,and BCL2.Result Mature follicles appeared after electroacupuncture intervention,the level of FSH went down(P<0.01),and the level of E_(2)went up(P<0.01).The oxidative stress level and the concentration of Fe^(2+)in the ovary of POF mice increased(P<0.01),and electroacupuncture up-regulated the level of SOD and down-regulated the levels of MDA and Fe^(2+)(P<0.01).The mRNA and protein expression of ferritin and GPX4 increased significantly after electroacupuncture treatment(P<0.01).Besides,electroacupuncture also boosted the expression of BCL2 and inhibited the m RNA and protein expression of BAX and P53(P<0.01).Conclusion Electroacupuncture can produce satisfactory results in treating POF,which may result from inhibiting ferroptosis and apoptosis by targeting GPX4 of system Xc-.