Sun Zi-Xue, Qiu Quan, He Xin, Zhang Fang, Zhao Pei-Pei
Department of Reproductive Medicine, Henan Provincial Hospital of Traditional Chinese Medicine, Zhengzhou, Henan 450002, China.
Henan University of Chinese Medicine, Zhengzhou, Henan 450000, China.
Zhonghua Nan Ke Xue. 2022 May;28(5):437-444.
To study the effect of Yishen Tongluo Prescription (YTP) on the testis tissue of the male rats with oligoasthenospermia (OAS) and its action mechanism based on the PI3K-AKT-mTOR pathway.
We randomly divided 48 SPF male SD rats into 8 groups: normal control, OAS model control, L-carnitine (LC), high-dose YTP, low-dose YTP, Omipalisib inhibitor (OI), OI + high-dose YTP, and OI + low-dose YTP, with 6 rats in each group. We established a model of OAS in the latter 7 groups by intragastric administration of tripterygium wilfordii polyside, followed by intervention with corresponding drugs. After treatment, we obtained semen parameters from the rats, observed pathological changes in the testis tissue by HE staining, and determined the expressions of the PI3K-AKT-mTOR signaling pathway-related proteins and mRNA by Western blot and real-time fluorescence quantitative PCR (qRT-PCR).
Sperm concentration and total sperm motility were significantly improved in the LC and YTP groups compared with those in the OAS model control group (P < 0.001). HE staining showed irregularly arranged spermatogenic cells and narrowed lumina and widened gaps of seminiferous tubules in the OAS model controls, as well as similar pathological changes in the LC, YTP and OI + YTP groups. Significant up-regulation was observed in the protein expressions of p-Akt, CatSper-1 and HSPA2 in the LC group (P < 0.05), those of p-Akt, mTOR, catsper-1 and HSP2 in the low-dose YTP group (P < 0.05) and that of PI3K in the high-dose YTP group (P < 0.05) compared with those in the model controls. There were no statistically significant differences in the expressions of PI3K, mTOR and catsper-1 between the OI and model control groups (P > 0.05), nor in those of PI3K, p-Akt, mTOR, CatSper-1 and HSPA2 between the OI + YTP and the former two groups (P > 0.05). The mRNA expressions of PI3K, mTOR, CatSper-1 and HSPA2 were remarkably higher in the LC and YTP groups than in the model control (P< 0.05), with those of catsper-1 and PI3K even more significantly up-regulated in the high-dose than in the low-dose YTP group (P< 0.001; P< 0.05). Statistically significant differences were not observed in the mRNA expressions of PI3K, mTOR, CatSper-1 and HSPA2 between the model control and OI groups (P > 0.05), nor in those of PI3K, mTOR, catsper-1 and HSPA2 between the model control and OI + YTP groups (P > 0.05).
Yishen Tongluo Prescription can improve sperm quality and pathological changes of the testis tissue in rats with Tripterygium glycoside-induced OAS, which might be attributed to its ability of up-regulating the expressions of the PI3K Akt mTOR pathway-related proteins and mRNA in the testis tissue.
基于PI3K-AKT-mTOR通路研究益肾通络方（YTP）对少弱精子症（OAS）雄性大鼠睾丸组织的影响及其作用机制。
将48只SPF级雄性SD大鼠随机分为8组：正常对照组、OAS模型对照组、左旋肉碱（LC）组、高剂量YTP组、低剂量YTP组、奥米帕利西布抑制剂（OI）组、OI+高剂量YTP组、OI+低剂量YTP组，每组6只。后7组通过灌胃雷公藤多苷建立OAS模型，随后给予相应药物干预。治疗后，获取大鼠精液参数，通过HE染色观察睾丸组织病理变化，采用蛋白质印迹法和实时荧光定量PCR（qRT-PCR）检测PI3K-AKT-mTOR信号通路相关蛋白和mRNA的表达。
与OAS模型对照组相比，LC组和YTP组的精子浓度和精子总活力显著提高（P<0.001）。HE染色显示，OAS模型对照组生精细胞排列不规则，生精小管管腔狭窄、间隙增宽，LC组、YTP组和OI+YTP组也有类似病理变化。与模型对照组相比，LC组中p-Akt、CatSper-1和HSPA2的蛋白表达显著上调（P<0.05），低剂量YTP组中p-Akt、mTOR、catsper-1和HSP2的蛋白表达上调（P<0.05），高剂量YTP组中PI3K的蛋白表达上调（P<0.05）。OI组与模型对照组之间PI3K、mTOR和catsper-1的表达无统计学差异（P>0.05），OI+YTP组与前两组之间PI3K、p-Akt、mTOR、CatSper-1和HSPA2的表达也无统计学差异（P>0.05）。LC组和YTP组中PI3K、mTOR、CatSper-1和HSPA2的mRNA表达显著高于模型对照组（P<0.05），高剂量YTP组中catsper-1和PI3K的mRNA表达上调幅度比低剂量YTP组更显著（P<0.001；P<0.05）。模型对照组与OI组之间PI3K、mTOR、CatSper-1和HSPA2的mRNA表达无统计学差异（P>0.05），模型对照组与OI+YTP组之间PI3K、mTOR、catsper-1和HSPA2的mRNA表达也无统计学差异（P>0.05）。
益肾通络方可以改善雷公藤多苷诱导的OAS大鼠的精子质量和睾丸组织病理变化，这可能与其上调睾丸组织中PI3K Akt mTOR通路相关蛋白和mRNA的表达有关。
