摘要
目的探讨强精片对睾丸支持细胞(SC)Toll样受体(TLRs)信号通路的作用。方法制备解脲支原体(UU)感染SC模型,分为模型组(空白血清)、强精片组(强精片含药血清)、TAK242组(TLR4抑制剂TAK242),另设正常SC采用空白血清培养为空白组。采用RT-q PCR和Western Blot分别检测TLR4、胞内下游分子髓样分化因子(My D88)、肿瘤坏死因子受体相关因6(TRAF6)、核因子κB(NF-κB)的mRNA和蛋白表达水平,免疫荧光技术检测NF-κB的核转位情况。结果与空白组比较,模型组TLR4、MyD88、TRAF6、NF-κB mRNA和蛋白表达、p NF-κB蛋白表达以及NF-κB核转位比例增加(P<0.05)。与模型组比较,强精片组TLR4、TRAF6 mRNA及蛋白表达降低,MyD88、NF-κB、p NF-κB蛋白表达及NF-κB核转位比例降低(P<0.05);TAK242组MyD88、TRAF6 mRNA及蛋白表达、NF-κB及pNF-κB蛋白表达及NF-κB核转位比例降低(P<0.05)。与TAK242组比较,强精片组TRAF6 mRNA表达升高,pNF-κB蛋白表达及NF-κB核转位比例降低(P<0.05)。结论强精片可抑制TLR4/MyD88/TRAF6/NF-κB信号通路,作用与TLR4抑制剂TAK242相近。
Objective To explore the effects of Qiangjing tablets(QJT)for Toll-like receptor signaling pathway(TLR)in testicular support cells(SC).Methods SC infected with Ureaplasma urealyticum(UU)model was established,and SC was divided into the model group(blank serum),QJT group(QJT containing serum),TAK242 group(TLR4 inhibitor TAK242).In addition,normal SC was cultured with blank serum as blank group.The mRNA and protein expression of TLR4,myeloid differentiation factor88(MyD88),tumor necrosis factor receptor-associated factor 6(TRAF6)、nuclear factor kappa-B(NF-κB)were detected by RTqPCR and Western Blot.The nuclear translocation of NF-κB was detected by immunofluorescence technique.Results Compared with blank group,TLR4,MyD88,TRAF6,NF-κB mRNA and protein expression,pNF-κB protein expression,and nuclear transposition ratio of NF-κB increased in model group(P<0.05).Compared with model group,TLR4,TRAF6 mRNA and protein expression,MYD88,NF-κB,pNF-κB protein expression and nuclear transposition ratio of NF-κB decreased in the QJT group(P<0.05).Compared with model group,MYD88 and TRAF6 mRNA and protein expression decreased,NF-κB and pNF-κB protein expression,and nuclear transposition ratio of NF-κB decreased in TAK242 group(P<0.05).Compared with TAK242group,TRAF6 mRNA expression increased,pNF-κB protein expression and nuclear transposition ratio of NF-κB decreased in QJT group(P<0.05).Conclusion QJT can inhibit the expression of TLR4,MyD88,TRAF6,NF-κB signaling pathway,the effect is similar to TLR inhibitor TAK242.
作者
尤耀东
张玲
朱坤
俞旭君
李广森
黄晓朋
常德贵
YOU Yao-dong;ZHANG Ling;ZHU KUN;YU Xu-jun;LI Guang-sen;HUANG Xiao-peng;CHANG De-gui(Key Laboratory of TCM Regulation of Metabolic Diseases of Sichuan Province,Affiliated Hospital of Chengdu University of Chinese Medicine,Chengdu 610075;Department of Andrology,Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072;Chengdu University of Traditional Chinese Medicine Clinical Medical College,Chengdu 610075;Department of Chinese Medicine Surgery,Reproductive Maternal and Child Hospital,Chengdu University of Traditional Chinese Medicine,Chengdu 610041)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2022年第11期1363-1368,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然基金面上项目资助(No.81673808,No.81973647)
成都市科技局项目基金资助(No.2015-HM01-00201-SF)
四川省科技厅(No.2019YJ0327)
四川省中医药管理局科技项目(No.2018QN010)。
关键词
强精片
TOLL样受体
睾丸支持细胞
不育
中药
Qiangjing tablets
Toll-like receptor
sertoli cell
infertility
Chinese herbal medicine
