益脾养胃汤治疗溃疡性结肠炎相关癌变小鼠的疗效及其机制研究

Therapeutic effect and mechanism of Yipi Yangwei decoction in treating ulcerative colitis-related cancerous mice

目的:观察益脾养胃汤治疗溃疡性结肠炎相关癌变小鼠的疗效,及其对TLR4/MAPKS信号通路的影响。方法:80只BALB/c雄性小鼠随机分为正常组,模型组,益脾养胃汤高、低剂量组,每组各20只。采用AOM/DSS法进行溃疡性结肠炎相关癌变小鼠模型的制备。造模后同时给予相应剂量的益脾养胃汤治疗。观察各组小鼠一般情况、结肠隐窝变化、结肠组织病理学的变化。采用ELISA法检测各组小鼠血清中炎性细胞因子IFN-γ,IL-4和IL-12的水平变化。Real-time PCR法检测IFN-γ,IL-4、IL-12及NF-κBp65 mRNA的表达。Western blot检测TLR4、p-JNK1/2、p-p38、pERK1/2、NF-κB (p65)、p-NF-κB (p65)、PCNA、COX-2、Bcl-xl蛋白的相对表达量。结果:与模型组比较,各给药组小鼠体质量和结肠长度明显增长,结肠重量、结肠重量长度比值和结肠黏膜组织学损伤评分明显降低;血清IFN-γ、IL-12水平和结肠组织中IFN-γ,IL-12 mRNA的表达量均明显降低,IL-4水平明显升高;结肠组织中TLR4、pJNK1/2、p-p38、p-ERK1/2的mRNA和蛋白表达量和p-NF-κBp65、PCNA、COX-2、Bcl-xl蛋白的表达量均显著降低(P均<0. 05)。与益脾养胃汤低剂量组比较,益脾养胃汤高剂量组结肠黏膜组织学损伤评分显著降低,各项因子水平显著优于低剂量组(P均<0. 05)。各组之间NF-κB(p65)的蛋白表达量比较差异无统计学意义(P>0. 05)。结论:益脾养胃汤可显著改善结肠黏膜的炎性损伤,减少癌瘤的形成,其机制可能与下调TLR4/MAPKS信号通路的表达有关。

AIM: To observe the therapeutic effect of Yipi Yangwei decoction in treating ulcerative colitis-related cancerous mice and its effect on toll-like receptor 4/mitogen-activated protein kinase( TLR4/MAPKS) signaling pathway. METHODS:Eighty BALB/c male mice were randomly divided into the normal group,the model group,the highdose and low-dose groups of Yipi Yangwei decoction. Each group contains 20 cases. AOM/DSS( azoxymethane/dextran sodium sulfate) method was used to establish murine mouse models of ulcerative colitis-associated cancer. After modeling,the corresponding dosage of Yipi Yangwei decoction was given to the models at the same time. The general condition,colon crypt changes,and colon histopathology changes of the mice in each group were observed.ELISA method was used to detect the levels of inflammatory cytokines IFN-γ,IL-4 and IL-12 in serum of each group. Real-time PCR was used to detect the expression of IFN-γ,IL-4,IL-12 and NF-κBp65 mRNA. The relative expression levels of TLR4,pJNK1/2 and p proteins were detected by Western blot. RESULTS: Compared with the model group,the body mass and colon length of the mice in each medication administration group were significantly increased, while the colon weight, colon weight length ratio,and colonic mucosal histological damage score were remarkably decreased; the levels of serum IFN-γ and IL-12,as well as the expression levels of IFN-γ and IL-12 mRNA in colon tissues were all significantly decreased,while IL-4 level was significantly increased; the mRNA and protein expression levels of TLR4,p-JNK1/2,p-p38 and pERK1/2 in colon tissues and the protein expression levels of p-NF-κBp65,PCNA,COX-2,Bcl-xl were significantly decreased( P < 0. 05). Compared with the low-dose group of Yipi Yangwei decoction,the histological injury score of colonic mucosa in the high-dose group of Yipi Yangwei decoction decreased significantly,and the levels of any other elements were all higher( P < 0. 05). The difference of protein expression levels of p-NF-κB( p65) in all groups was without statistical significance( P >0. 05). CONCLUSION: Yipi Yangwei decoction can relieve the inflammatory injury of colonic mucosa and reduce the risk of cancer. The mechanism may be related to the down-regulation of TLR4/MAPKS signaling pathway.