沙苑子总黄酮对雄性生殖功能障碍模型大鼠的改善作用及机制研究
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篇名: 沙苑子总黄酮对雄性生殖功能障碍模型大鼠的改善作用及机制研究
TITLE: Study on Improvement Effects and Mechanism of Astragalus complanatus Total Flavonoids on Reproductive Dysfunction Model Male Rats
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,2g/kg)和TFA低、中、高剂量组(0.05、0.1、0.2g/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35d。给药结束后,将各组雄性大鼠与雌性大鼠按1∶1合笼交配7d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。
ABSTRACT: OBJECTIVE: To stu dy 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 E 2 and improving oxidant stress level.
期刊: 2020年第31卷第15期
作者: 刘雯,侯晨辉,李江
AUTHORS: LIU Wen,HOU Chenhui ,LI Jiang
关键字: 沙苑子总黄酮;生殖功能障碍;性激素;氧化应激;机制;大鼠
KEYWORDS: Astragalus complanatus total flavonoids ;Reproductive dysfunction ;Horone;Oxidant stress ;Mechanism;Rat
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