补肾调经方对体外培养小鼠卵母细胞线粒体功能及其与卵丘颗粒细胞缝隙连接的影响  

Effects of Bushen Tiaojing Formula on mitochondria function and gap connection with granulosa cells in mouse oocytes in vitro

作　　者：吕金梦 耿静然 张拴成 梁潇 杜惠兰 LYU Jin-meng;GENG Jing-ran;ZHANG Shuan-cheng;LIANG Xiao;DU Hui-lan(College of Integrated Traditional Chinese and Western Medicine,Hebei University of Chinese Medicine,Institute of Integrative Medicine,Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,Shijiazhuang 050091,China;Hebei Province Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou 061000,China;Hebei Maternity Hospital,Shijiazhuang 050091,China)

机构地区：[1]河北中医学院中西医结合学院,中西医结合研究所,河北省中西医结合肝肾病证研究重点实验室,石家庄050091 [2]河北省沧州中西医结合医院,沧州061000 [3]河北生殖妇产医院,石家庄050091

出　　处：《中华中医药杂志》2023年第1期131-135,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774359)。

摘　　要：目的:探讨补肾调经方对体外培养小鼠卵母细胞线粒体功能及其与卵丘颗粒细胞缝隙连接的影响。方法:50只SD大鼠随机分为补肾组和血清对照组,每组25只,分别灌胃补肾调经方和蒸馏水,采血并分离血清后得到补肾组血清和血清对照组血清。将270只雌性ICR小鼠随机分为补肾组、血清对照组、空白对照组,每组90只。分别用含有补肾组血清、血清对照组血清和不含大鼠血清的培养液培养卵泡至成熟,收集MⅡ期卵母细胞计算卵母细胞第一极体排出率,活细胞荧光探针观察线粒体均匀分布比率,Western Blot法检测UCP2、PGC-1α蛋白表达;收集卵丘颗粒细胞,通过流式细胞术和Western Blot法分别检测细胞凋亡率和缝隙连接蛋白(Cx)43、Cx37蛋白表达。结果:补肾组卵母细胞第一极体排出率较其余两组显著升高(P<0.05)。补肾组卵丘颗粒细胞凋亡率较其余两组显著降低(P<0.05)。血清对照组和补肾组Cx37、Cx43蛋白表达均较空白对照组显著降低(P<0.05),补肾组Cx37、Cx43蛋白表达较血清对照组显著升高(P<0.05)。补肾组PGC-1α、UCP2蛋白表达较其余两组显著升高(P<0.05)。结论:补肾调经方可通过改善线粒体功能和减少卵丘颗粒细胞凋亡,增加Cx,从而提高卵母细胞质量。Objective:To explore the the effects of Bushen Tiaojing Formula on mitochondria function and gap connection with cumulus granulosa cells in vitro cultured mouse oocytes.Methods:A total of 50 SD rats were randomly divided into Bushen group and serum control group with 25 in each group and fed Bushen Tiaojing Formula and distilled water respectively,then collected blood and separated serum,serum of Bushen group and serum control group were obtained.A total of 270 female ICR mice were randomized into Bushen group,serum control group,and blank control group with 90 mice in each group.Follicles were cultured until maturity with culture medium containing Bushen group serum,serum control group serum and no serum and collecting MI oocytes,to observe the mitochondrial distribution with live cell fluorescent probes,and the protein expression of UCP2 and PGC-1αwas detected by Weston Blot;The cumulus granulosa cells were collected and apoptosis rate and protein expression of gap junction proteins Cx43 and Cx37 were determined by flow cytometry and Western Blot,respectively.Results:The rate of first pole discharge of oocytes in the Bushen group was significantly higher than that in the remaining two groups(P<0.05).The apoptosis rate of ovarian mound granulosa cells in the kidney tonifying group was significantly reduced compared with the other two groups(P<0.05).Cx37 and Cx43 protein expression in the serum control group and Bushen group was significantly reduced compared with the blank control group(P<0.05),and the expression of Cx37 and Cx43 proteins in the Bushen group was significantly increased compared with the serum control group(P<0.05).The expression of PGC-1αand UCP2protein was significantly increased in the Bushen group(P<0.05).Conclusion:Bushen Tiaojing Formula can improve the quality of oocytes by improving mitochondrial function and reducing granulosate apoptosis and increasing of gap connexin.

关 键 词：补肾调经方 卵母细胞 体外成熟培养 颗粒细胞 线粒体 

分 类 号：R285.5[医药卫生—中药学]