人参皂苷Rg1减轻热应激致小鼠睾丸损伤的机制研究  

Mechanism of ginsenoside Rg1 alleviating testicular injury induced by heat stress in mice

作　　者：黄寅虎 汪子铃 杜坤航 王程 黄彩虹 杨婷[1] 魏晗 蒋鸿慧 王璐[1] 张庆华 王亚平[1] HUANG Yinhu;WANG Ziling;DU Kunhang;WANG Cheng;HUANG Caihong;YANG Ting;WEI Han;JIANG Honghui;WANG Lu;ZHANG Qinghua;WANG Yaping(Department of Histology and Embryology,Laboratory of Stem Cell and Tissue Engineering,College of Basic Medical Sciences,Chongqing Medical University,Chongqing,400016;Department of Gynaecology and Obstetrics,Army Medical Center of PLA,Chongqing,400042,China)

机构地区：[1]重庆医科大学:干细胞与组织工程研究室,组织学与胚胎学教研室,重庆400016 [2]陆军特色医学中心妇产科,重庆400042

出　　处：《陆军军医大学学报》2024年第10期1123-1131,共9页Journal of Army Medical University

摘　　要：目的探讨人参皂苷Rg1减轻热应激致小鼠睾丸损伤的机制。方法20只6~8周龄雄性C57BL/6J小鼠按随机数字表法分为4组,每组5只。对照组(Control组)和热应激组(HS组)小鼠腹腔注射0.9%生理盐水[10 mL/(kg·d)×14 d],热应激+人参皂苷Rg1组(HS+Rg1组)和人参皂苷Rg1组(Rg1组)小鼠腹腔注射人参皂苷Rg1[20 mg/(kg·d)×14 d]。HS组与HS+Rg1组小鼠在给药第7天时,将麻醉后小鼠下腹部置于43℃水浴中单次热应激30 min。小鼠精母细胞GC-2spd(ts)分为4组:对照组(Control组)细胞常规培养48 h;热应激组(HS组)细胞常规培养36 h,置于43℃水浴中单次热应激30 min,继续培养12 h;热应激+Rg1组(HS+Rg1组)在细胞培养体系中加入Rg1(终浓度50μmol/L),热应激处理同HS组;Rg1组不给予热应激处理,其余同HS+Rg1组。建模完成后1 d采集眼球血,ELISA法检测血清睾酮水平;摘取睾丸观察形态并称量,计算睾丸指数,HE染色观察睾丸组织病理学形态;制备睾丸组织匀浆检测过氧化氢酶(CAT)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平;取附睾精子,计算机辅助精子分析(CASA)系统分析小鼠精子浓度和活力;TUNEL染色检测GC-2spd(ts)细胞凋亡情况;Western blot检测GC-2spd(ts)细胞Nrf2、Keap1、HO-1、Bax、Bcl-2和Caspase3蛋白表达量;RT-qPCR检测GC-2spd(ts)细胞Nrf2蛋白靶基因GCLC、GCLM和NQO1相对表达量。结果Rg1可阻止热应激所致睾丸质量和睾丸指数下降,减轻睾丸组织结构损伤,阻止精子浓度和活力下降,拮抗热应激所致睾丸间质细胞数量减少和血清睾酮水平下降,降低睾丸组织中MDA累积,提高抗氧化酶CAT和SOD的活性。Rg1还可减轻GC-2spd(ts)细胞凋亡,下调Bax、Caspase3和Keap1蛋白表达,增强Bcl-2、Nrf2和HO-1蛋白表达量,增强Nrf2蛋白靶基因GCLC、GCLM和NQO1相对表达量。结论Rg1对正常小鼠睾丸结构和功能影响不显著,但可有效提高小鼠睾丸抗热应激损伤能力,其机制可能与Rg1激活Nrf2信号通路,提�Objective To explore the mechanism of ginsenoside Rg1 in alleviating heat stress-induced testicular injury in mice.Methods A total of 20 C57BL/6 male mice(6~8 weeks old)were randomly divided into 4 groups(n=5).The mice from the control group and heat stress(HS)group were intraperitoneally injected with 10 mL/(kg·d)0.9%normal saline for 14 d,while those in the HS+Rg1 group and the Rg1 group were given an intraperitoneal injection of 20 mg/(kg·d)for 14 d,and then on the 7th day after administration,the mice in the HS group and the HS+Rg1 group had the lower abdomen put into a 43℃water bath for 30 min as a single heat stress after being anesthetized with 4%chloral hydrate.Mouse spermatocytes GC-2spd(ts)were divided into control group(routine culture for 48 h),HS group(placed in a 43℃water bath for 30 min after 36 h of conventional culture,and cultured till the end of 48 h),HS+Rg1 group(50μmol/L Rg1 treatment followed by heat stress injury),and Rg1 group(no heat stress injury).In 1 d after modeling,the eyeball blood samples were collected to detect serum testosterone with ELISA,and the testicles were extracted to observe the morphology and weighed to calculate the testicular index.HE staining was used to observe the histopathology of testis,and corresponding reagents and kits was employed to detect the content of malondialdehyde(MDA)and activities of catalase(CAT)and superoxide dismutase(SOD)in testis tissue.After the epididymal sperm were collected,the sperm concentration and motility were analyzed by computer-assisted sperm analysis(CASA)system.In in vitro experiments,cell apoptosis was detected with TUNEL staining,the protein levels of Nrf2,Keap1,HO-1,Bax,Bcl-2 and Caspase3 were detected with Western blotting,and the mRNA levels of GCLC,GCLM and NQO1 were detected by RT-qPCR.Results Rg1 prevented the decreases in testicular weight and testicular index caused by heat stress,reduced the damage of testicular tissue structure,prevented the decrease of sperm concentration and vitality,antagonized the decreasd nu

关 键 词：人参皂苷RG1 睾丸 热应激 睾酮 氧化应激损伤 机制 

分 类 号：R282.71[医药卫生—中药学] R339.21[医药卫生—中医学] R339.52