“秩边透水道”针法对卵巢低反应小鼠外源性凋亡因子表达的影响  

The effect of“Zhibian(BL 54)-to-Shuidao(ST 28)”acupuncture technique on the expression of exogenous apoptotic factors in mice with poor ovarian response

作　　者：任佳 常博雅 崔梦洁 高珍 曹玉霞 王海军 冀来喜 REN Jia;CHANG Bo-ya;CUI Meng-jie;GAO Zhen;CAO Yu-jxia;WANG Hai-jjun;JI Lai-xi(Shanxi University of Traditional Chinese Medicine,Shanxi Jinzhong 030619,China)

机构地区：[1]山西中医药大学,山西晋中030619

出　　处：《时珍国医国药》2024年第10期2486-2493,共8页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82074549);山西省中医药管理局基金(2023ZYYB048);山西省教育厅基金(2023L187)。

摘　　要：目的观察“秩边透水道”针法对卵巢低反应小鼠(poor ovarian response,POR)外源性凋亡因子表达的影响,探讨针刺对卵巢功能的改善作用。方法将30只动情周期规律的C57BL/6雌性小鼠随机分为空白组、模型组和针刺组(n=10)。除空白组外,余下组别均构建POR模型。造模成功后,对针刺组行针刺干预。其余组只行固定操作。干预结束次日3组同时促排,次日取材。观察各组小鼠一般状况并记录处死前小鼠的体质量;采用阴道脱落细胞涂片法观察小鼠的动情周期变化;检测小鼠的获卵数、卵巢湿重与卵巢指数;ELISA法检测血清中AMH、FSH、E_(2)、LH的含量;HE染色法观察卵巢形态结构;TUNEL染色法检测卵巢组织阳性细胞凋亡率;RT-PCR法和Western Blot法分别检测卵巢组织中Fas、FasL、FADD、TRAIL、DR5、Caspase-8、Caspase-3 mRNA和蛋白表达。结果与空白组比较,模型组小鼠动情周期紊乱率升高(P<0.01);获卵数、卵巢湿重和卵巢指数降低(P<0.01);血清中FSH和LH含量升高(P<0.01),AMH和E_(2)含量下降(P<0.01);卵巢组织病理形态变差;阳性细胞凋亡率提高(P<0.01);Fas、FasL、FADD、TRAIL、DR5、Caspase-8、Caspase-3 mRNA表达水平及蛋白相对表达量升高(P<0.01,P<0.05)。与模型组比较,针刺组小鼠动情周期紊乱率降低(P<0.01);获卵数、卵巢湿重和卵巢指数提高(P<0.01);血清中FSH和LH含量降低(P<0.01),AMH和E_(2)含量升高(P<0.01);卵巢组织形态明显改善;阳性细胞凋亡率降低(P<0.01);Fas、FasL、FADD、TRAIL、DR5、Caspase-8、Caspase-3 mRNA表达水平及蛋白相对表达量降低(P<0.01,P<0.05)。结论“秩边透水道”针法可能通过调节外源性凋亡通路中的关键因子,改善POR小鼠卵巢功能障碍,减少卵巢颗粒细胞凋亡,提高卵巢反应性。Objective By observing the effect of“Zhibian(BL 54)-to-Shuidao(ST 28)”acupuncture on the expression of exogenous apoptotic factors in poor ovarian response(POR)mice,the improvement of acupuncture on ovarian function was discussed.Methods Thirty C57BL/6 female mice with regular estrous cycle were randomly divided into blank group,model group and acupuncture group(n=10).The model group and the acupuncture group were given tripterygium glycosides suspension(50 mg·kg^(-1))by gavage for 14 days,and the blank group was given an equal volume of 0.9%sodium chloride solution for 14 days,once a day for 20 min.After the modeling was completed,the acupuncture group was treated with“Zhibian(BL 54)-to-Shuidao(ST 28)”acupuncture for 14 days.The blank group was only fixed without other interventions.The three groups were treated at the same time,and the corresponding indicators were detected the next day.The general condition of mice in each group was observed and the body weight of mice before death was recorded;the changes of estrous cycle in mice were observed by vaginal exfoliated cell smear;the number of eggs,ovarian wet weight and ovarian index of mice were detected.The levels of anti-mullerian hormone(AMH),follicle-stimulating hormone(FSH),estradiol(E_(2))and luteinizing hormone(LH)in serum were detected by enzyme-linked immunosorbent assay(ELISA);Hematoxylin-eosin(HE)staining was used to observe the morphological structure of ovary;the apoptosis rate of positive cells in ovarian tissue was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining;the mRNA and protein expressions of Fas cell surface death receptor(Fas),FasL,fas-associated death domain(FADD),tumor necrosis factor-related apoptosis-inducing ligand(TRAIL),death receptor 5(DR5),cysteine-aspartic acid protease(Caspase)-8 and Caspase-3 in ovarian tissues were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-PCR)and western blot(WB),respectively.Results Compared wit

关 键 词：“秩边透水道” 卵巢低反应 针刺 凋亡 

分 类 号：R246[医药卫生—针灸推拿学]