首页|"秩边透水道"针刺对弱精子症小鼠睾丸内外凋亡途径关键因子的影响

"秩边透水道"针刺对弱精子症小鼠睾丸内外凋亡途径关键因子的影响

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目的 观察"秩边透水道"针刺对弱精子症模型小鼠生殖功能的影响,并探讨可能作用机制.方法 将42只C57BL/6雄性小鼠随机分成空白组、模型组和针刺组各14只.模型组和针刺组小鼠给予环磷酰胺30 mg/(kg·d)腹腔注射7天建立弱精子症小鼠模型,空白组腹腔注射7天0.9%氯化钠溶液10 ml/(kg·d).造模成功后,针刺组小鼠给予"秩边透水道"针刺,每日 1次,每次20 min,每周6次,共3周;其余两组只进行固定而不针刺.日常观察各组小鼠的一般情况,并记录干预前后体质量变化.干预结束次日每组随机取6只雄鼠检测精子质量以及睾丸、附睾质量并计算相应指数;ELISA法检测血清中睾酮(T)、促卵泡生成素(FSH)、促黄体生成激素(LH)含量;HE染色和TUNEL染色观察睾丸和附睾病理组织形态和细胞凋亡情况;荧光定量PCR法和Western blot法分别检测睾丸组织死亡相关因子(Fas)、Fas相关死亡域蛋白(FADD)、B淋巴细胞瘤2(Bcl-2)、B淋巴细胞瘤2关联X的蛋白(Bax)和半胱氨酸蛋白酶3(Caspase-3)mRNA和蛋白表达变化.将每组剩余的8只雄鼠与处于动情周期的同品系雌鼠进行1∶1合笼,检测合笼后每组小鼠的妊娠率和每胎胚胎数量.结果 与空白组比较,模型组小鼠体重减轻,睾丸质量、睾丸指数、附睾质量和附睾指数下降,精子总数、精子活力和精子活率下降,血清T、FSH和LH含量下降,细胞凋亡率升高,Fas、FADD、Bax和Caspase-3 mRNA和蛋白表达水平提高,Bcl-2 mRNA和蛋白表达水平降低(P<0.01),睾丸组织和附睾组织病理形态异常,生精小管排列紊乱,管腔内生精细胞数量减少;合笼后雌鼠的妊娠率和每胎胚胎数量降低(P<0.01).与模型组比较,针刺组小鼠睾丸质量、睾丸指数、附睾质量、附睾指数、精子总数、精子活力和精子活率均提高,血清T、FSH和LH含量提高,细胞凋亡量降低,Fas、FADD、Bax和Caspase-3 mRNA和蛋白表达水平降低,Bcl-2 mRNA和蛋白表达水平提高(P<0.05或P<0.01),睾丸组织和附睾组织形态改善,生精小管排列紧密,管腔内生精细胞数量增加;合笼后雌鼠的妊娠率和每胎胚胎数量提高(P<0.01).结论 "秩边透水道"针刺可以改善弱精子症模型小鼠睾丸组织细胞凋亡,提高生殖功能,其机制可能与调节睾丸组织外在膜受体途径(Fas介导的途径)和内在线粒体途径(Bcl-2/Bax调控的途径)的关键因子有关.
Effect of Acupuncture at"Zhibian(BL 54)-to-Shuidao(ST 28)"on Key Factors of Endogenous Exogenous Apoptotic Pathways in Asthenospermia Model Mice
Objective To observe the effect of acupuncture"Zhibian(BL 54)-to-Shuidao(ST 28)"on the re-productive function of asthenospermia model mice and to explore the possible mechanism.Methods Forty-two male C57BL/6 mice were randomly divided into blank group,model group,and acupuncture group,with 14 mice in each group.Cyclophosphamide 30 mg/(kg·d)was injected intraperitoneally for 7 days to establish model of asthenosper-mia for the model group and the acupuncture group,while 0.9%sodium chloride solution 10 ml/(kg·d)was injected intraperitoneally for 7 days in the blank group.After successful modeling,mice in the acupuncture group received acupuncture at"Zhibian(BL 54)-to-Shuidao(ST 28)"once a day for 20 minutes,6 times a week for 3 weeks;the re-maining two groups were fixed without acupuncture.Daily observations were made on the general conditions of mice in all groups,and changes in body weight after intervention were recorded.On the next day after completing the treat-ment,6 male mice selected randomly from each group to test the sperm quality as well as testicular and epididymal weights,and calculate the corresponding indices;ELISA was used to test the levels of testosterone(T),follicle-stimulating hormone(FSH),and luteinizing hormone(LH)in serum;HE staining and TUNEL staining were per-formed to observe the pathological morphology and apoptosis of testicular and epididymal tissues;fluorescence quantitative PCR and western blot were used to detect changes in the expression of apoptosis-related factors(Fas),Fas-associated death domain protein(FADD),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and cysteine-aspartic acid protease(caspase)-3 mRNA and protein in testicular tissue.The remaining 8 male mice in each group were housed with estrus-cycling mice of the same strain at 1:1 ratio,and the pregnancy rate and number of embryos per litter in each group were determined after mating.Results In comparison with the blank group,mice in the model group exhibited reduced body weight,decreased testicular mass,testicular index,epididymal mass,and epididymal index.Additionally,there was a decrease in total sperm count,sperm motility,and sperm viability.Serum levels of T,FSH,and LH were reduced.The apoptosis rate increased,and the expression levels of Fas,FADD,Bax,and Caspase-3 mRNA and proteins were elevated,while Bcl-2 mRNA and protein expression levels decreased(P<0.01).Pathological abnormalities were observed in testicular and epididymal tissues,with disrupted arrangement of seminiferous tubules and a decreased number of sperniatogenic cells within the tubular lumen.Furthermore,the pregnancy rate and the number of embryos per litter decreased significantly after mating(P<0.01).In comparison with the model group,mice in the acupuncture group showed improvements in testicular mass,testicular index,epididymal mass,epididymal index,total sperm count,sperm motility,and sperm viability.Serum levels of T,FSH,and LH increased.The apoptosis rate decreased,and the expression levels of Fas,FADD,Bax,and Caspase-3 mRNA and proteins decreased,while Bcl-2 mRNA and protein expression levels increased(P<0.05 or P<0.01).Morphological improvements were observed in testicular and epididymal tissues,with a compact arrangement of seminiferous tubules and an increased number of spermatogenic cells within the tubular lumen.Fur-thermore,the pregnancy rate and the number of embryos per litter increased significantly after mating(P<0.01).Conclusion Acupuncture"Zhibian(BL 54)-to-Shuidao(ST 28)"can improve testicular tissue apoptosis and enhance reproductive function in a mouse model of asthenospermia.Its mechanism may be associated with the modula-tion of key factors in the extrinsic membrane receptor pathway(Fas-mediated pathway)and the intrinsic mitochondrial pathway(Bcl-2/Bax-regulated pathway)in testicular tissue.

asthenospermiaZhibian(BL 54)Shuidao(ST 28)acupunctureapoptosis

郝健亨、任佳、常博雅、王海军、冀来喜

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成都中医药大学针灸推拿学院,四川省成都市金牛区十二桥路37号,610075

山西中医药大学

弱精子症 秩边 水道 针刺 细胞凋亡

国家自然科学基金山西省中医药管理局科研课题山西中医药大学2020年度科技创新能力培育计划

820745492023ZYYB0482020PY-LP-03

2024

中医杂志
中华中医药学会 中国中医科学院

中医杂志

CSTPCD北大核心
影响因子:1.464
ISSN:1001-1668
年,卷(期):2024.65(1)
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