摘要
目的:研究沙苑子总黄酮(TFA)对腺嘌呤诱导的雄性生殖功能障碍模型大鼠的改善作用及机制。方法:将雄性SD大鼠分为空白组、模型组、阳性组(五子衍宗丸,2 g/kg)和TFA低、中、高剂量组(0.05、0.1、0.2 g/kg),每组12只。各组雄性大鼠每日早上9:00灌胃相应药物,并于同日下午3:00灌胃腺嘌呤混悬液以复制生殖功能障碍模型(空白组除外),连续给药35 d。给药结束后,将各组雄性大鼠与雌性大鼠按1∶1合笼交配7 d后取出,检测致雌鼠受孕率;采用酶联免疫吸附法检测雄性大鼠血清中睾酮(T)、雌二醇(E2)、促卵泡激素(FSH)、促黄体激素(LH)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)、肌酐(Scr)、尿素氮(BUN)水平;摘取雄性大鼠睾丸、附睾、精囊腺、胸腺、肾,并计算其脏器指数;采用苏木精-伊红(HE)染色法观察雄性大鼠睾丸、附睾及肾组织的病理学变化;采用免疫组织化学染色检测雄性大鼠肾组织中B细胞淋巴瘤/白血病2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶3(Caspase-3)、Caspase-9的蛋白表达水平。结果:与空白组比较,模型组雄性大鼠致雌鼠受孕率,睾丸、附睾、精囊腺、胸腺指数,血清中T、E2、SOD、GSH-Px水平,肾组织中Bcl-2蛋白表达水平均显著降低(P<0.01);肾指数和血清中FSH、LH、MDA、Scr、BUN水平,肾组织中Bax、Caspase-3、Caspase-9蛋白表达水平均显著升高(P<0.01);睾丸组织中生精小管直径变细、间距较宽,生精细胞排列紊乱;附睾管管壁增厚、排列疏松、间质增大,管腔增生萎缩、腔内精子聚集成团;肾组织中肾间质纤维增生,肾小管内可见大量黄色结晶。与模型组比较,阳性组和TFA中、高剂量组雄性大鼠致雌鼠受孕率,睾丸、附睾、精囊腺、胸腺指数,血清中T、E2、SOD、GSH-Px水平,肾组织中Bcl-2蛋白表达水平均显著升高(P<0.05或P<0.01),肾指数和血清中FSH、LH、MDA、Scr、BUN水平,肾组织中Bax、Caspase-3、Caspase-9蛋白表达水平均显著降低(P<0.05或P<0.01);TFA低剂量组上述指标变化趋势相同,且大部分有显著性差异(P<0.05或P<0.01);各给药组雄性大鼠睾丸、附睾、肾组织病理学变化均明显减轻。结论:TFA对腺嘌呤诱导的雄性大鼠的生殖障碍具有显著改善作用,其机制可能与降低性激素FSH、LH水平和下调肾组织中凋亡相关蛋白表达,升高性激素T、E2水平和改善机体氧化应激水平有关。
OBJECTIVE: To study the improvement effects of Astragalus complanatus total flavonoids(TFA) on adenine-induced reproductive dysfunction model male rats and its mechanism. METHODS:Male SD rats were divided into blank group,model group,positive group(Wuzhi yanzong pill,2 g/kg),TFA high-dose,medium-dose and low-dose groups(0.05,0.1,0.2 g/kg),with 12 rats in each group. Male rats in each group were given the corresponding drugs intragastrically at 9:00 a.m.every day,while the model of reproductive dysfunction was reproduced by intragastric administration of adenine suspension(except for blank group)at 3:00 p.m. on the same day,for consecutive 35 days. After medication,male rats in each group were mated with female rats by ratio of 1 ∶ 1 in cages;7 days later,inducing-pregnancy rate of male rats were measured. ELISA assay was used to detect the serum levels of T,E2,FSH,LH,GSH-Px,SOD,MDA,Scr and BUN in male rats;the testis,epididymis,seminal vesicle,thymus and kidney of male rats were extracted and the organ index was calculated;HE staining was used to observe the pathological changes of testis,epididymis and kidney;the protein expression of Bcl-2,Bax,Caspase-3 and Caspase-9 in the renal tissue of male rats were detected by immunohistochemistry. RESULTS:Compared with blank group,the inducing-pregnancy rate of male rats,the testis,epididymis,seminal vesicle gland,thymus indexes,the serum levels of T,E2,SOD and GSH-Px,the protein expression levels of Bcl-2 in renal tissue were reduced significantly in model group(P<0.01);the renal index,the serum levels of FSH,LH,MDA,Scr and BUN,the protein expression levels of Bax,Caspase-3 and Caspase-9 in renal tissue were increased significantly(P<0.01);the diameter of seminiferous tubules in testis became thinner,the spacing was wider,and the arrangement of seminiferous cells was disordered;the wall of epididymis was thickened,the arrangement was loose,the stroma was enlarged,the lumen was proliferated and atrophied,and the sperm in the lumen wass aggregated;in renal tissue,interstitial fibers was proliferated,and a large number of yellow crystals were seen in renal tubules. Compared with model group,the inducing-pregnancy rate,the testis,epididymis,seminal vesicle gland,thymus indexes,the serum levels of T,E2,SOD and GSH-Px,the protein expression levels of Bcl-2 in renal tissue of male rats were increased significantly in the positive group,TFA medium-dose and high-dose groups(P<0.05 or P<0.01),while the renal index,the serum levels of FSH,LH,MDA,Scr and BUN,the protein expression levels of Bax,Caspase-3 and Caspase-9 in renal tissue were decreased significantly(P<0.05 or P<0.01);there was the same chage trend and most with significant difference in above indexes of TFA low-dose group(P<0.05 or P<0.01);the pathalogic changes of testis, epididymis and renal tissue were improved significantly.CONCLUSIONS:TFA can significantly improve adenine-induced reproductive dysfunction model male rats. The mechanism may be associated with reducing hormone FSH and LH level,the expression of apoptosis-related proteins in renal tissue,while increasing the level of hormone T and E2 and improving oxidant stress level.
作者
刘雯
侯晨辉
李江
LIU Wen;HOU Chenhui;LI Jiang(Nursing College,Zhengzhou Railway Vocational and Technical College,Zhengzhou 451460,China;Henan Natural Medicine Extraction and Medical Technology Application Engineering Research Center,Zhengzhou 451460,China)
出处
《中国药房》
CAS
北大核心
2020年第15期1854-1860,共7页
China Pharmacy
基金
河南省科技发展计划项目(No.202102310088)
河南省高等学校重点科研项目(No.20A320086)。
关键词
沙苑子总黄酮
生殖功能障碍
性激素
氧化应激
机制
大鼠
Astragalus complanatus total flavonoids
Reproductive dysfunction
Horone
Oxidant stress
Mechanism
Rat
