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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(P<0.01),HSP70蛋白和mRNA表达均显著降低(P<0.01)。与模型组比较,各给药组结肠黏膜损伤评分均显著降低(P<0.01),各给药组均可增加结肠组织HSP70的蛋白和mRNA表达(P<0.05,P<0.01),其中以健脾益肠散高剂量组最为明显(P<0.05,P<0.01)。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。
Objective: To investigate the effects of Jianpi Yiganchang on the expression of heat shock protein 70 (HSP70) in colonic tissue of rats with ulcerative colitis (UC). Methods: Sixty SPF male Sprague-Dawley rats were randomly divided into 2 groups: normal group and model group. UC model was established by using TNBS / The model group was randomly divided into 5 groups: model group, sulfasalazine group (0.3 g · kg -1), and spleen and intestine group in high, medium and low dose groups (204, 136, 68 g · kg ~ (-1)) for 10 days. After the rats were orally administered with the corresponding drugs for 21 days, the general state of the rats and the damage of colonic mucosa tissues were observed. Immunohistochemistry and Western blot ) And real-time PCR (Real-time PCR) were used to detect the protein and mRNA expression of HSP70 in rat colon. Results: Compared with the normal group, the colon mucosal lesion score of the model group was significantly increased (P <0.01), while the expression of HSP70 protein and mRNA were significantly decreased (P <0.01). Compared with the model group, the colon mucosal lesion score of each administration group was significantly decreased (P <0.01), and the protein and mRNA expression of HSP70 in the colon tissue of all the treated groups were increased (P <0.05, P <0.01) Spleen and intestine scattered high dose group was the most obvious (P <0.05, P <0.01). Conclusion: Jianpi Yigan San may play a therapeutic role by promoting HSP70 expression in colonic mucosa of UC rats.