摘要
目的探讨移植骨髓间充质干细胞(BMSC)来源的Leydig样细胞对邻苯二甲酸二(2-乙基己基)酯(DEHP)染毒小鼠睾丸功能的修复作用及其可能机制。方法将6只C57小鼠随机分为DEHP组与对照组,进行DEHP染毒28 d后,获取小鼠血清行睾酮检测;同时获取睾丸组织行HE染色及TUNEL染色,并检测睾丸半胱氨酸蛋白酶(caspase)-3、caspase-8、caspase-9、Bcl-2和Bax蛋白变化情况,测定丙二醛(MDA)、谷胱甘肽(GSH)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性。同时,将27只C57小鼠进行DEHP染毒28 d,随机分为Leydig样细胞移植组、BMSC移植组及对照组(n=9),分别将BMSC来源的Leydig样细胞、BMSC移植到相应组别C57小鼠睾丸中,在移植后7、14、21 d时每组选取3只小鼠血液检测血清睾酮水平;同时获取双侧睾丸,检测睾丸组织MDA、GSH含量及SOD、GSH-Px活性。移植后7 d时获取小鼠睾丸行HE染色及TUNEL染色,同时检测caspase-3、caspase-8、caspase-9、Bcl-2及Bax蛋白表达情况。结果与对照组比较,DEHP组C57小鼠血清睾酮水平明显下降,睾丸生精细胞紊乱,组织细胞出现凋亡,凋亡蛋白caspase-3、caspase-8、caspase-9及Bax蛋白表达水平明显增高,Bcl-2蛋白表达水平明显降低,MDA含量升高,GSH含量降低,GSH-Px及SOD活性受到抑制,差异均有统计学意义(P<0.05或P<0.01)。行细胞移植后,与对照组比较,移植后7、14、21 d时Leydig样细胞移植组小鼠血清睾酮水平明显升高,移植后14、21 d时BMSC移植组血清睾酮水平明显升高,在移植后7、14、21 d,Leydig样细胞移植组及BMSC移植组睾丸组织MDA含量明显降低,GSH含量明显升高,GSH-Px及SOD活性恢复(P<0.05或P<0.01);移植后7 d,Leydig样细胞移植组及BMSC移植组睾丸组织细胞凋亡较对照组明显减轻,生精细胞形态有所恢复,同时caspase-3、caspase-8、caspase-9及Bax蛋白表达水平明显降低,而Bcl-2蛋白表达水平明显升高(P<0.05或P<0.01)。结论移�
Objective To investigate the effect and mechanism of transplanting Leydig-like cells from BMSCs to repair the testicular function of diethylhexyl phthalate(DEHP)infected mice.Methods Six C57 mice were randomly divided into two groups(3 each):control group(corn oil)and DEHP group[900 mg/(kg.d)DEHP mixed with corn oil].Venous blood was taken from the eyeball 28 days after treatment to determine the serum testosterone level.One testis was stored in paraformaldehyde for HE and TUNEL staining,the other side of testis,after homogenization,was used to determine the changes of apoptotic proteins of caspase-3,caspase-8,caspase-9,Bcl-2 and Bax with Western blotting,the contents of MDA and GSH and the activities of SOD and GSH-Px were determined.Twenty-seven C57 mice were poisoned with DEHP for 28 days,and then randomly divided into Leydig-like cell transplant group,BMSCs transplant group and control group(9 each).BMSCs and Leydig-like cells from BMSCs were transplanted into the testis of mice in corresponding groups.Venous blood was taken from 3 mice of each group at 7th,14th and 21st day after transplantation for determining the serum testosterone,and bilateral testis were obtained simultaneously to determine the contents of MDA and GSH and the activities of SOD and GSH-Px.The testis of mice was got 7th day after transplantation for HE and TUNEL staining,and the changes of apoptotic proteins of caspase-3,caspase-8,caspase-9,Bcl-2 and Bax were detected simultaneously.Results Compared with control group,the serum testosterone level decreased significantly,testicular spermatogenic cell disorder,tissue cells showed apoptosis,the serum levels of apoptotic protein caspase-3,caspase-8,caspase-9 and Bax increased obviously,while of Bcl-2 decreased markedly in testis of DEHP group,the content of MDA increased,and of GSH decreased,the activities of GSH-Px and SOD were inhibited with statistically significant difference(P<0.05 or P<0.01).Compared with control group,the serum testosterone level in Leydig-like cell transplantation gro
作者
康永明
方琨
宋攀
杨璐辰
刘正欢
王临春
史率克
董强
Kang Yong-Ming;Fang Kun;Song Pan;Yang Lu-Chen;Liu Zheng-Huan;Wang Lin-Chun;Shi Shuai-Ke;Dong Qiang(Department of Urology,West China Hospital of Sichuan University,Chengdu 610041,China;Department of Urology,Suining Central Hospital,Suining,Sichuan 627000,China)
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2021年第8期749-756,共8页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金面上项目(81771569)。
