大黄蛰虫丸抑制坏死性凋亡信号通路改善大鼠睾丸衰老的作用和机制  

Effect and mechanism of Dahuang Zhechong Pills against testicular aging in rats by inhibiting necroptosis signaling pathway

作　　者：李欢[1] 涂玥 万毅刚[4] 穆耕林 吴薇[4] 陈佳鑫 王美子 王杰[1] 符燕 蔡玉丰 王玉[1] WAN Zi-yue LI Huan;TU Yue;WAN Yi-gang;MU Geng-lin;WU Wei;CHEN Jia-xin;WANG Mei-zi;WANG Jie;FU Yan;CAI Yu-feng;WANG Yu;WAN Zi-yue(Deparment of Traditional Chinese Medicine,Nanjing Drum Touer Hospial Clinical Cllege of Nanjing Uniersity of Chinese MedicineNanjing 20008,China;Acupuncture and Moxibustion and Massage Colge&Health Preservation and Rehabilitation College,Nanjing Universiy of Chinese Medicine,Nanjing 20023,China;Jiangsu Provuincial TCM Technology Engineering Research Cenerof Heath and Heath Preseration,Nanjing Unersity,Chinse Medicine,Nanjing 210003 China;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospial,The Afiliated Hospital of Nanjing University Medical School,Nanjing 210008,China;Institute of Chinese Medicine,Nanjing University,Nanjing 210008,China;Graduate School of Social Sciences,Faculty of Social Sciences,Hitotsubashi University,Tokyo 186-8601,Japan)

机构地区：[1]南京中医药大学鼓楼临床医学院中医科,江苏南京210008 [2]南京中医药大学针灸推拿学院·养生康复学院,江苏南京210023 [3]江苏省中医药健康养生技术工程研究中心,江苏南京210023 [4]南京大学医学院附属鼓楼医院中医科,江苏南京210008 [5]南京大学中医研究院,江苏南京210008 [6]Graduate School of Social Sciences Faculty of Social Sciences Hitotsubashi University,Tokyo 186-8601

出　　处：《中国中药杂志》2022年第15期4119-4127,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82174472,81573903);江苏省自然科学基金面上项目(BK20211298);江苏省中医药科技发展计划项目(MS2021037);江苏省研究生科研与实践创新计划项目(SJCX21_0672,KYCX21_1699);中央高校基本科研业务费专项资金项目(021414380518);南京市医学科技发展项目(YKK20085);南京市名中医专家工作室建设项目。

摘　　要：为了探究传统经方——大黄蛰虫丸(Dahuang Zhechong Pills,DHZCP)在体内改善睾丸衰老(testicular aging,TA)的作用和机制,笔者将24只雄性大鼠随机分成正常组、模型组、DHZCP组和维生素E(vitamin E,VE)组,通过D-半乳糖(D-galactose,D-gal)连续灌胃建立大鼠TA模型。在实验期间,每天经灌胃分别给予DHZCP组和VE组大鼠DHZCP混悬液和VE混悬液;此外,经灌胃给予正常组和模型组大鼠生理盐水。在D-gal和不同药物联合干预60 d后,处死全部大鼠,收集血液及睾丸组织,进而,针对模型鼠TA和睾丸细胞坏死性凋亡相关的各项指标进行检测和观察。这些指标包括各组大鼠衰老表型、睾丸总质量和睾丸指数、睾丸组织病理学特征和生精细胞数、性激素水平、睾丸组织活性氧(reactive oxygen species,ROS)产物表达特征、睾丸组织细胞周期蛋白表达水平和表达特征以及受体相互作用蛋白激酶1(receptor interacting serine/threonine protein kinase 1,RIPK1)/受体相互作用蛋白激酶3(receptor interacting serine/threonine protein kinase 3,RIPK3)/底物混合谱系激酶样蛋白(mixed lineage kinase domain like pseudokinase,MLKL)信号通路关键信号分子蛋白表达水平。结果显示,对于TA模型鼠,DHZCP和VE都能改善衰老表型、睾丸总质量和睾丸指数、睾丸组织病理性特征和生精细胞数、血清睾酮和血清卵泡刺激素水平、睾丸组织ROS表达特征以及P21、P53蛋白表达水平和表达特征;此外,DHZCP与VE都能改善模型鼠睾丸组织RIPK1/RIPK3/MLKL信号通路关键信号分子蛋白表达水平;其中,对于RIPK3的改善作用,DHZCP优于VE。总之,在该研究中,笔者发现,DHZCP与VE相似,在体内能改善D-gal诱导的大鼠TA,其作用机制与抑制RIPK1/RIPK3/MLKL信号通路活性而减轻睾丸细胞坏死性凋亡有关。该研究为传统经方在男性健康领域的开发应用提供了初步的药理学证据。To explore the effect and mechanism of Dahuang Zhechong Pills(DHZCP),a classical prescription,in improving testicular aging(TA)in vivo,the authors randomly divided 24 male rats into four groups:the normal,model,DHZCP and vitamin E(VE)groups.The TA rat model was established by continuous gavage of D-galactose(D-gal).During the experiment,the rats in the DHZCP and VE groups were given DHZCP suspension and VE suspension,respectively by gavage,while those in the normal and model groups were gavaged saline separately every day.After the co-administration of D-gal and various drugs for 60 days,all rats were sacrificed,and their blood and testis were collected.Further,various indexes related to TA and necroptosis of testicular cells in the model rats were examined and investigated,which included the aging phenotype,total testicular weight,testicular index,histopathological features of testis,number of spermatogenic cells,sex hormone level,expression characteristics of reactive oxygen species(ROS)in testis,expression levels and characteristics of cyclins in testis,and protein expression levels of the key molecules in receptor-interacting serine/threonine-protein kinase 1(RIPK1)/receptor-interacting serine/threonine-protein kinase 3(RIPK3)/mixed lineage kinase domain like pseudokinase(MLKL)signaling pathway in each group.The results showed that,for the TA model rats,both DHZCP and VE improved their aging phenotype,total testicular weight,testicular index,pathological features of testis,number of spermatogenic cells,serum testosterone and follicle stimulating hormone levels,expression characteristics of ROS and protein expression levels and characteristics of P21 and P53 in testis.In addition,DHZCP and VE improved the protein expression levels of the key molecules in RIPK1/RIPK3/MLKL signaling pathway in testis of the model rats.Specifically,DHZCP was better than VE in the improvement of RIPK3.In conclusion,in this study,the authors found that DHZCP,similar to VE,ameliorated D-gal-induced TA in model rats in vivo,and its m

关 键 词：大黄蛰虫丸 睾丸衰老 坏死性凋亡 RIPK1/RIPK3/MLKL信号通路 

分 类 号：R285.5[医药卫生—中药学]