人脐带间充质干细胞上调Nrf2信号改善衰老小鼠睾丸功能  

Human umbilical cord mesenchymal stem cells improve testicular function in aging mice by upregulating Nrf2 signaling

作　　者：王圆圆 薛函 李林艳 刘娟 吴雨瞳 何青 余丽梅 Wang Yuanyuan;Xue Han;Li Linyan;Liu Juan;Wu Yutong;He Qin;Yu Limei(Guizhou Province Key Laboratory of Cell Engineering,Affiliated Hospital of Zunyi Medical University Guizhou Provincial Prenatal Diagnosis Sub-center(Affiliated Hospital of Zunyi Medical University)Engineering Research Center for Guizhou Clinical Application of Stem Cells and Drug Research and Development(Zunyi Medical University),Zunyi 563003,China;Collaborative Innovation Center of Tissue Damage Repair and Regenerative Medicine of Ministry of Education,Zunyi Medical University,Zunyi 563003,China)

机构地区：[1]遵义医科大学附属医院贵州省细胞工程重点实验室、贵州省产前诊断分中心、贵州省干细胞临床应用与药物开发工程研究中心(遵义医科大学),遵义563003 [2]遵义医科大学教育部组织损伤修复与再生医学协同创新中心,遵义563003

出　　处：《中华生殖与避孕杂志》2024年第6期579-586,共8页Chinese Journal of Reproduction and Contraception

基　　金：国家自然科学基金(82360047);遵义医科大学附属医院博士后基金(rc220330427)。

摘　　要：目的探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)移植对氧化损伤衰老小鼠睾丸功能的影响及分子机制。方法共纳入18只6~8周SPF级C57BL/C雄性小鼠,完全随机分组法分为3组,对照组:小鼠注射等量生理盐水;模型组:颈背部皮下注射D-半乳糖连续9周,造模第4周末,每只小鼠尾静脉注射生理盐水;hUCMSCs组:颈背部皮下注射D-半乳糖连续9周,造模第4周末,每只小鼠尾静脉注射hUCMSCs。9周后,称取小鼠体质量和睾丸重量,计算睾丸指数,酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测小鼠血清睾酮水平、睾丸组织丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)含量。肉眼观察睾丸外观,苏木精-伊红(hematoxylin-eosin,HE)染色观察睾丸组织病理学变化,实时荧光定量聚合酶链反应和Western blotting分别检测核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)信号转导相关基因和蛋白表达。结果模型组小鼠睾丸指数[(0.64±0.05)%]较对照组[(0.81±0.13)%,P=0.006]降低,睾丸体积缩小,生精小管完整性破坏,生精细胞和精子减少,间质稀疏;血清睾水平[(4.10±0.67)μg/L]、睾丸组织SOD活性[(48.87±6.40)U/mg Prot]较对照组[(5.71±0.81)μg/L,P=0.002;(78.53±9.70)U/mg Prot,P=0.001]均降低,而MDA含量[(1.11±0.19)nmol/mg Prot]较对照组[(0.77±0.07)nmol/mg Prot,P=0.001]增加。Keap1 mRNA和蛋白表达量较对照组均增高(P=0.006,P=0.043),Nrf2、SOD和NQO1 mRNA表达量较对照组减少(P=0.002,P<0.001,P=0.001),Nrf2、HO-1蛋白表达量较对照组均明显降低(P=0.011,P=0.021)。与模型组比较,hUCMSCs组小鼠睾丸指数[(0.79±0.03)%,P=0.010]增高;睾丸组织结构较为清晰、完整,生精小管各级生精细胞和精子及间质细胞较丰富;血清睾酮水平[(5.24±0.21)μg/L,P=0.028]及睾丸组织SOD活性[(79.47±14.32)U/mg Prot,P=0.001]增高,MDA含量[(0.77±0.08)nmol/mg Prot,P=0.001]降低;NObjective To investigate the effect and mechanism of human umbilical cord mesenchymal stem cells(hUCMSCs)transplantation on testicular function in aging mice with oxidative damage.Methods Totally 18 SPF grade C57BL/C male mice aged 6-8 weeks were randomly divided into 3 groups using a complete randomization method.In control group,mice were injected with an equal amount of physiological saline;in model group,mice were subcutaneous injected D-galactose into the neck and back for 9 consecutive weeks,on the 4th weekend of modeling,the mice were injected with physiological saline via the tail vein;in hUCMSCs group:mice were subcutaneous injected D-galactose into the neck and back for 9 consecutive weeks,on the 4th weekend of modeling,the mice were injected with hUCMSCs via the tail vein of each mouse.After 9 weeks,body weight and testicular weight of the three groups mice were measured and testicular index was calculated.The contents of testosterone,malondialdehyde(MDA)and superoxide dismutase(SOD)in serum and testicular tissue were detected by enzyme-linked immunosorbent assay(ELISA)method.Visual observation of testicular appearance,the histopathological changes of testis were observed by hematoxylin-eosin(HE)staining,and the expression of NF-E2-related factor 2(Nrf2)signal transduction-related genes and proteins were detected by RT-PCR and Western blotting,respectively.Results Compared with control group[(0.81±0.13)%],the testicular index of mice in model group[(0.64±0.05)%,P=0.006]was decreased.In model group,the volume of testis was reduced,the integrity of spermatogenic tubules was damaged,spermatogenic cells and sperm were reduced,and the interstitium was sparse.In model group,serum testosterone[(4.10±0.67)μg/L]and SOD[(48.87±6.40)U/mg Prot]were decreased compared with control group[(5.71±0.81)μg/L,P=0.002;(78.53±9.70)U/mg Prot,P=0.001],MDA[(1.11±0.19)nmol/mg Prot]was increased compared with control group[(0.77±0.07)nmol/mg Prot,P=0.001],Keap1 mRNA and protein expression were increased(P=0.006,P=0.043

关 键 词：睾丸 核因子E2相关因子2 衰老 人脐带间充质干细胞 D-半乳糖 抗氧化 

分 类 号：R965[医药卫生—药理学]