首页|期刊导航|针刺研究|"秩边"透"水道"对卵巢低反应小鼠卵巢组织内/外源性凋亡相关因子表达的影响

"秩边"透"水道"对卵巢低反应小鼠卵巢组织内/外源性凋亡相关因子表达的影响OA北大核心

Effect of penetrative needling of"Zhibian"(BL54)through"Shuidao"(ST28)on expression of internal and external apoptosis-related factors in ovarian of mice with poor ovarian response

中文摘要英文摘要

目的:观察"秩边"透"水道"针刺对卵巢低反应(POR)小鼠内/外源性凋亡相关因子的影响,探讨针刺对POR小鼠卵巢功能的保护机制.方法:C57BL/6N雌鼠按随机数字表法分为空白组、模型组和针刺组,每组23只.采用雷公藤多苷片混悬液(50 mg·kg-1)灌胃制备POR模型.针刺组采用"秩边"透"水道"针刺,每次20 min,每日1次,连续干预14 d.选取30只同品系7~8周龄健康雄鼠与部分雌鼠进行1∶1合笼.阴道脱落细胞涂片观察雌鼠的动情周期变化;观察获卵数,测定卵巢湿重并计算卵巢指数;观察合笼后的妊娠率和胚胎数量;ELISA法检测雌鼠血清低水平抗米勒管激素(AMH)、促卵泡生成素(FSH)、促黄体生成激素(LH)、雌二醇(E2)含量;HE染色法观察卵巢组织形态;TUNEL染色观察卵巢细胞凋亡率;实时荧光定量PCR法和Western blot法分别检测卵巢组织半胱氨酸蛋白酶(Caspase)-3、Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、细胞因子受体超家族成员6(Fas)、凋亡相关蛋白调节因子(FADD)mRNA和蛋白表达水平.结果:与空白组相比,模型组小鼠动情周期紊乱率升高(P<0.01),妊娠率和胚胎数量明显减少(P<0.01),获卵数、卵巢湿重、卵巢指数明显降低(P<0.01);血清中FSH和LH含量升高(P<0.01),AMH和E2含量下降(P<0.01);卵巢细胞凋亡率明显升高(P<0.01),卵巢组织中闭锁卵泡明显增加、各级卵泡明显减少且颗粒细胞排列紊乱;卵巢组织中Caspase-3、Bax、Fas、FADD mRNA和蛋白表达水平升高(P<0.01),Bcl-2 mRNA和蛋白表达水平降低(P<0.01).与模型组相比,针刺组小鼠动情周期紊乱率下降(P<0.01),妊娠数和胚胎数量增多(P<0.01),获卵数、卵巢湿重和卵巢指数增加(P<0.01);血清中FSH和LH含量下降(P<0.01),AMH和E2含量上升(P<0.01);卵巢细胞凋亡率明显降低(P<0.01),卵巢颗粒细胞排列整齐,闭锁卵泡减少、各级卵泡数量增加;卵巢组织中Caspase-3、Bax、Fas、FADD mRNA和蛋白表达水平降低(P<0.01,P<0.05),Bcl-2 mRNA和蛋白表达水平升高(P<0.01,P<0.05).结论:"秩边"透"水道"针刺可以改善POR小鼠卵巢反应降低,其机制可能与调节卵巢组织内/外源性凋亡相关因子的表达有关.

Objective To observe the effect of penetrative needling of"Zhibian"(BL54)through"Shuidao"(ST28)on the expressions of internal and external apoptosis-related factors in mice with poor ovarian response(POR),so as to explore its mechanisms underlying improvement of POR.Methods Sixty-nine female C57BL/6N mice were randomly divided into blank,model and acupuncture groups(n=23 in each group).The POR model was established by gavage of tripterygium wilfordii polyglucoside suspension(50 mg·kg-1).After successful modeling,the rats in the acupuncture group received penetrative needling of BL54 through ST28,with the needle retained for 20 min,once a day for 14 days.Thirty healthy male mice of the same strain aged 7-8 weeks were co-housed with female mice in a 1∶1 ratio.Vaginal exfoliative cell smears were used to observe the changes in the estrous cycle of female mice.The number of follicles were observed under light microscope,wet weight of ovaries was recorded and ovarian index was calculated.The pregnancy rate and number of embryos were observed.ELISA method was used to detect the low levels of anti Mullerian hormone(AMH),follicle stimulating hormone(FSH),luteinizing hormone(LH),and estradiol(E2)contents in the serum of female mice.HE staining was used to observe the morphology of ovarian tissue.TUNEL staining was used to observe the apoptosis rate of ovarian cells.The mRNA and protein expression levels of cysteine-aspartic acid protease(Caspase)-3,Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),fas cell surface death receptor(Fas),and fas-associated death domain(FADD)in ovarian tissue were detected by real-time quantitive PCR and Western blot,respectively.Results Compared with the blank group,the rate of estrous cycle disorder,the contents of FSH and LH in serum,the rate of ovarian apoptosis,the mRNA and protein expression levels of Caspase-3,Bax,Fas and FADD in ovarian tissues were significantly increased(P<0.01);whereas the pregnancy rate,the number of embryos,the number of eggs obtained,ovarian wet weight,ovarian index,the contents of AMH and E2,the mRNA and protein expression levels of Bcl-2 were significantly decreased(P<0.01)in the model group.Following the intervention,the above-mentioned indexes were all reversed in the acupuncture group(P<0.01,P<0.05).Conclusion Penetrative needling of BL54 through ST28 can increase the ovarian response in POR mice,which may be associated with its function in regulating the expression of internal and external apoptosis-related factors in ovarian tissue.

任佳;常博雅;崔梦洁;高珍;曹玉霞;王海军

山西中医药大学第二临床学院,山西 晋中 030600山西中医药大学第二临床学院,山西 晋中 030600山西中医药大学第二临床学院,山西 晋中 030600山西中医药大学第二临床学院,山西 晋中 030600山西中医药大学第二临床学院,山西 晋中 030600山西中医药大学第二临床学院,山西 晋中 030600

卵巢低反应"秩边"透"水道"针刺细胞凋亡

Poor ovarian response"Zhibian"(BL54)through"Shuidao"(ST28)AcupunctureApoptosis

《针刺研究》 2025 (2)

123-130,140,9

国家自然科学基金面上项目(No.82074549)山西省自然科学基金面上项目(No.201901D111399)山西中医药大学博士科研启动基金项目(No.2023BK17)山西省教育厅项目(No.2023L187)

10.13702/j.1000-0607.20231108

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