五子衍宗丸对肾精亏虚证大鼠细胞色素C氧化酶7a2及细胞外信号调节激酶磷酸化水平表达的影响

Effects of Wuzi Yanzong Pill on the expression of Cox7a2 gene and ERK phosphorylation in rats with kidney essence deficiency pattern

目的观察五子衍宗丸对肾精亏虚证大鼠睾丸组织细胞色素C氧化酶7a2(Cox7a2)表达和细胞外信号调节激酶(ERK)磷酸化水平的影响。方法 SD雄性大鼠随机分为5组,即空白组,模型组,五子衍宗丸低、中、高剂量组,每组5只。空白组每天给予羧甲基纤维素钠灌胃,模型组和五子衍宗丸组每天给予雷公藤多甙(20 mg/kg)灌胃,五子衍宗丸组分别给予五子衍宗丸低、中、高剂量(0.5,1.0,2.0 g/kg)灌胃30 d,第31天取材。采用荧光定量PCR检测Cox7a2表达,Western Blot法检测ERK磷酸化水平表达,Johnsen’s评分评价睾丸组织病理学变化。结果雷公藤多甙可诱导Cox7a2 mRNA表达增加及ERK磷酸化,同时睾丸生精功能下降,Johnsen’s评分降低。与模型组相比,五子衍宗丸高剂量组ERK磷酸化水平下降,五子衍宗丸中、高剂量组Cox7a2 mRNA水平明显下调,并且五子衍宗丸中、高剂量组睾丸组织Johnsen’s评分显著上调,睾丸生精功能改善。结论五子衍宗丸可通过ERK信号传导途径调节线粒体能量代谢,从而改善生精功能。

Objective To observe the effects of Wuzi Yanzong Pill（ Five-Seed Progeny Pill,WZYZP） on Cox7a2 gene and extracellular signal-regulated kinase（ ERK） phosphorylation level in rats with kidney essence deficiency pattern. Methods Sprague-Dawley（ SD） male rats were randomly divided into five groups（ n = 5 in each group） ： blank control group,model group,low-dose WZWZP（ WZWZP-L）group,mid-dose WZWZP（ WZWZP-M） group,and high-dose WZWZP（ WZWZP-H）. The blank control group were given sodium carboxy methyl cellulose（ Na CMC） via intragastric gavage. The model group and three WZWZP groups received intragastric gavage of Leigongteng（ Tripterygium Wilfordii） glycosides（ GTW） alone（ 20 mg / kg） or GTW followed by WZWZP（ low dose of 0. 5g / kg,medium dose of 1.0g / kg,or high dose of 2. 0 g / kg daily） respectively for 30 consecutive days. Cox7a2 mRNA expression in testis was determined by using real-time reverse transcriptase polymerase chain reaction（ RTPCR）. The extracellular signal-regulated kinase1 /2（ ERK1 /2） phosphorylation level was assessed with Western blot assay. Histopathological changes in the testes were evaluated using Johnsen＇s scale. Results GTW induced an increase in Cox7a2 mRNA expression and an activation of ERK as well as a decline of Johnsen＇s score and significant decrease in testicular spermatogenesis. Compared with GTW group,ERK phosphorylation level in high-dose WZWZP group decreased markedly while high-dose and mid-dose WZWZP downregulated Cox7a2 mRNA levels. Furthermore,Johnsen＇s score in all WZWZP groups increased significantly. Conclusion WZWZP could protect the impaired spermatogenesis,probably by mediating mitochondrial energy metabolism via ERK pathway.