健脾益肠散对溃疡性结肠炎大鼠结肠组织HSP70表达的影响

目的:探讨健脾益肠散对溃疡性结肠炎(UC)大鼠结肠组织热休克蛋白70(HSP70)蛋白和mRNA表达的影响。方法:将健康SPF级雄性SD大鼠60只,随机分为2组,正常组和造模组;造模组采用二硝基苯磺酸(TNBS)/乙醇法复制UC大鼠模型;待复制模型成功后将造模组随机分为5组,分别为模型组、柳氮磺吡啶组(0.3 g·kg~(-1))以及健脾益肠散高、中、低剂量组(204,136,68 g·kg~(-1)),每组10只;灌胃相应药物21 d后,观察各组大鼠的一般状态和结肠黏膜组织损伤情况,免疫组化、蛋白免疫印迹法(Western blot)和实时荧光定量PCR(Real-time PCR)分别检测大鼠结肠组织中HSP70的蛋白和mRNA表达。结果:与正常组比较,模型组大鼠结肠黏膜损伤评分显著升高(P0.01),HSP70蛋白和mRNA表达均显著降低(P0.01)。与模型组比较,各给药组结肠黏膜损伤评分均显著降低(P0.01),各给药组均可增加结肠组织HSP70的蛋白和mRNA表达(P0.05,P0.01),其中以健脾益肠散高剂量组最为明显(P0.05,P0.01)。结论:健脾益肠散可能通过促进HSP70的表达而达到对UC大鼠结肠黏膜的免疫保护,从而发挥治疗作用。

Effect of Jianpi Yichang Powder on Expression Levels of HSP70 in Colon Tissues of Rats with Ulcerative Colitis

Objective: To explore the effect of Jianpi Yichang powder (JP) on protein and mRNA expression levels of heat shock protein 70 (HSP70) in colon tissues of rats with ulcerative colitis (UC). Method: The 60 healthy SPF male SD rats were randomly divided into normal group and model group. The UC models were established by trinitro-benzene-sulfonic acid(TNBS)/ethanol method. Then the successfully modeled rats were randomly divided into the model group, sulfasalazine group (0.3 g ·kg^-1), and JP high, middle and low dose groups (204, 136 and 68 g ·kg^-1) with 10 rats in each group. After ig administration for 21 days, the general condition and the colonic mucosa injury of rats were observed. Immunohistochemistry, Western blot and Real-time PCR were used to detect the protein and mRNA expression levels of HSP70 in colon tissues of rats. Result: As compared with the normal group, the colon mucosal injury score was increased significantly (P<0.01), and the protein and mRNA expression levels of HSP70 were significantly decreased (P<0.01) in model group. As compared with the model group, the colon mucosal injury scores in each drug group were significantly lower (P<0.01), and the protein and mRNA expression levels of HSP70 were significantly increased (P<0.05, P<0.01). The effect was most obvious in JP high dose group (P<0.05, P<0.01). Conclusion: JP may promote the expression of HSP70 to protect intestinal mucosal immunity in UC rats, so as to play a therapeutic role.