何首乌饮对运动疲劳大鼠睾丸组织P450胆固醇侧链裂解酶和类固醇激素急性调节蛋白的影响

Effects of Heshouwuyin on the expression of P450 side-chain cleavage enzyme and steroidogenic acute regulatory protein in testis tissue of the exercised-induced fatigue rat

目的探讨何首乌饮对运动疲劳大鼠睾丸组织细胞色素P450胆固醇侧链裂解酶(P450scc)和类固醇激素急性调节蛋白(StAR)的影响。方法 60只雄性SD大鼠,随机分为安静对照组(A组)、安静何首乌饮组(B组)、模型组(C组)、自然恢复组(D组)、何首乌饮治疗组(E组);何首乌饮预防组(F组),每组10只。C、D、E和F组大鼠复制运动疲劳动物模型,其中F组每天训练前给何首乌饮20g/(kg.d)(含生药4.8kg/L)灌胃60 d。模型成功后E组给何首乌饮20 g/(kg.d)(含生药4.8kg/L)灌胃治疗60 d。采用美国Beckmancoulter Unicel Dxl 800仪器测定睾酮水平;采用RT-PCR、Western blotting和免疫组织化学法检测各组睾酮合成限速酶P450scc和StAR的表达变化。结果 P450scc免疫组织化学阳性颗粒主要表达于间质细胞及精母细胞,B组和F组表达最强,C组最弱,与其余各组相比差异有统计学意义。P450scc蛋白和mRNA表达变化为B、F组明显高于E、D和A组(P<0.05),C组明显低于A组(P<0.05),A组和D组以及B组和F组两两之间差异无显著性;StAR免疫组织化学阳性颗粒主要表达于睾丸间质细胞胞质,阳性强度表现为E组和F组最强,C组最弱。StAR蛋白和mRNA表达变化为E和F组明显高于A、D组(P<0.05),C组明显低于A组(P<0.05),A、D组之间无差异。结论何首乌饮可以提高睾酮合成限速酶P450scc和StAR的表达。

Objective To explore the effect of Heshouwuyin on the expression of P450 side-chain cleavage enzyme（P450scc） and steroidogenic acute regulatory protein （STAR） in testis tissue of rats with exercised-induced fatigue. Methods Sixty SD rats were randomly divided into 6 groups, 10 rats in each group, including a normal control group （group A）, a Heshouwuyin administered normal group（ group B）, a model control group（ group C）, a spontaneously recovering group（ group D）, a Heshouwuyin treated group （group E）, and a Heshouwuyin prevented group （group F）. Groups C, D, E and F were exercised-induced fatigue animal model, groups in which animals were administrated Heshouwuyin [ 20g/（ kg. d） , contained crude drug 4. 8kg/L]. Animals in group F were administrated Heshouwuyin before modeling, and animals in group E were treated with Heshouwuyin for 60 days after success of modeling. Beckmancoulter Unicel Dxl 800 was used to detect the level of serum testosterone, according to the manufacture＇ s instructions. Western blotting and RT-PCR were used to observed the differential expression of P450scc and STAR. Result P450sce protein was expressed in Leydig ceils and spermatocyte. Compared with group A, the expression of P450sec protein and mRNA in groupC decreased （ p 〈 0.05） , and the expression of P450scc protein and mRNA in groups B and F increased more than E, D, A. StAR protein was expressed in Leydig cells. Compared with group A, the expression of StAR protein and mRNA in group C decreased （P 〈 0. 05） , and the expression of StAR protein and mRNA in groups E and F increased more than A, D, but there was no difference between groups A and D （P 〉 0. 05）. Conclusion The expression of P450scc and StAR may be up-regulated by Heshouwuyin.