【摘要】 目的:探讨人参皂苷Rg3对邻苯二甲酸二丁酯（DBP）诱导的雄性生殖功能损伤小鼠的生精保护作用,并阐明其作用机制。方法:30只6周龄清洁级C57BL/6小鼠随机分为对照组、DBP组(给予400 mg·kg^-1DBP)和DBP+Rg3组(给予400 mg·kg^-1DBP和20 mg·kg^-1Rg3),每组10只。实验期末处死小鼠并分析各组小鼠精子质量,HE染色观察各组小鼠睾丸组织形态表现,免疫组织化学法观察各组小鼠睾丸组织中间隙连接蛋白43 （Cx43）的表达,实时荧光定量PCR （RTqPCR）法检测各组小鼠睾丸组织中Src、磷脂酰肌醇-3-激酶（PI3K）和蛋白激酶B （Akt） mRNA表达水平,Western blotting法检测各组小鼠睾丸组织中磷酸化Src （p-Src）、磷酸化PI3K （p-PI3K）和磷酸化Akt （p-Akt）蛋白表达水平。结果:与对照组比较,DBP组小鼠的精子密度和精子活力均明显降低（P<0.01）;与DBP组比较,DBP+Rg3组小鼠精子密度和精子活力均明显升高（P<0.01）。HE染色,与DBP组比较,人参皂苷Rg3组小鼠睾丸组织病理学损伤有所改善。免疫组织化学法检测,与对照组比较,DBP组小鼠睾丸组织中Cx43蛋白表达量降低;与DBP组比较,DBP+Rg3组小鼠睾丸组织中Cx43蛋白表达量升高。RT-qPCR法检测,与对照组比较,DBP组小鼠睾丸组织中Src、PI3K和Akt mRNA的表达水平降低（P<0.01）;与DBP组比较,DBP+Rg3组小鼠睾丸组织中Src、PI3K和Akt mRNA表达水平升高（P<0.01）。Western blotting法检测,与对照组比较,DBP组小鼠睾丸组织中p-Src、p-PI3K和p-Akt蛋白表达水平降低（P<0.01）,与DBP组比较,DBP+Rg3组小鼠睾丸组织中p-Src、p-PI3K和p-Akt蛋白表达水平升高（P<0.01）。结论:人参皂苷Rg3对DBP诱导的小鼠生殖功能损伤具有改善作用,其机制可能是Rg3通过激活Src/PI3K/Akt通路,使Cx43表达上调,降低DBP暴露所致的血睾屏障的通透性,发挥生精损伤的保护作用。更多还原

【Abstract】 Objective:To investigate the protective effect of ginsenoside Rg3 on spermatogenesis in the male mice with reproductive function injury induced by dibutyl phthalate（DBP）,and to elucidate its mechanism. Methods:Thirty 6-week-old clean grade C57BL/6 mice were randomly divided into control group,DBP group(given 400 mg·kg^-1 DBP) and DBP+Rg3 group(given 400 mg·kg^-1 DBP and20 mg·kg^-1 Rg3)and there were 10 mice in each group. At the end of the experiment,the mice were sacrificed and the sperm quality was analyzed. The pathomorphology of testis tissue of the mice in various groups were observed by HE staining. The expression of gap junction connexin 43（Cx43）protein in testis tissue of the mice in various groups was observed by immunohistochemistry. The expression levels of Src,phosphatidylinositol-3-kinase（PI3K） and protein kinase B（Akt） mRNA in testis tissue of the mice in various groups were detected by Real-time quantitative PCR（RT-qPCR）. Western blotting method was used to detect the expression levels of phosphorylated Src（p-Src）,phosphorylated PI3K（p-PI3K） and phosphorylated Akt（p-Akt）proteins in testis tissue of the mice in various groups. Result:Compared with control group,the sperm density and sperm motility of the mice in DBP group were significantly decreased（P<0. 01）;compared with DBP group,the sperm density and sperm motility of the mice in DBP+Rg3 group were significantly increased（P<0. 01）. The HE staining results showed that compared with DBP group,the histopathological injury of testis tissue of the mice in ginsenoside Rg3 group was improved. The immunohistochemical analysis results showed that the expression amont of Cx43 protein in testis tissue of the mice in DBP group was lower than that in control group;compared with DBP group,the expression amount of Cx43 protein in testis tissue of the mice in DBP+Rg3 group was increased. The RT-qPCR results showed that compared with control group,the expression levels of Src,PI3K and Akt mRNA in testis tissue of the mice in DBP group were decreased（P<0. 01）;compared with DBP group,the expression levels of Src,PI3K and Akt mRNA in testis tissue of the mice in DBP+ Rg3 group were increased（P<0. 01）. The Western blotting detection results showed that compared with control group,the expressions levels of p-Src,p-PI3K,and p-Akt proteins in testis tissue of the mice in DBP group were decreased（P<0. 01）;compared with DBP group,the expressions levels of p-Src,p-PI3K,and p-Akt proteins in testis tissue of the mice in DBP+Rg3 group were increased（ P<0. 01）.Conclusion :Ginsenoside Rg3 can improve the DBP-induced reproductive function injury in the mice,and the mechanism may be that Rg3 can up-regulate the expression of Cx43 by activating the Src/PI3K/Akt pathway,reduces the permeability of blood-testosterone barrier induced by DBP exposure,and plays a protective role in the spermatogenic injury.更多还原