健脾化滞方对急性溃疡性结肠炎小鼠的治疗作用及对炎症因子的影响

Therapeutic effect of Jianpi Huazhi Formula on acute ulcerative colitis mice and its influence on inflammatory factors

目的:探讨健脾化滞方对急性溃疡性结肠炎(UC)模型小鼠的治疗作用,并基于小鼠血清及结肠组织炎症因子表达水平探讨其作用机制。方法:将76只健康雄性C57小鼠随机分为正常组、模型组、美沙拉秦组及健脾化滞方低、中、高剂量组。通过饮用3%葡聚糖硫酸钠(DSS)溶液建立急性UC小鼠模型,健脾化滞方低、中、高剂量组小鼠分别灌胃给予145.8、291.5、583.0 g/L健脾化滞方溶液,美沙拉秦组小鼠灌胃给予10 g/L美沙拉秦溶液。观察各组小鼠症状,进行疾病活动指数(DAI)评分,苏木素-伊红(HE)染色观察小鼠结肠组织病理变化,并采用速率法和终点法检测小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、直接胆红素(DBil)、总胆红素(TBil)、白蛋白(ALB)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(γ-GT)、总胆汁酸(TBA)、肌酐(Cre)、尿酸(UA)、尿素氮(BUN)水平,采用酶联免疫吸附实验(ELISA)测定小鼠血清与结肠组织中白介素(IL)-6、肿瘤坏死因子-α(TNF-α)、IL-10的水平。结果:健脾化滞方可有效改善急性UC小鼠便血、腹泻等临床症状,改善其体质量的下降及小鼠结肠长度的缩减,并能降低小鼠DAI评分(P<0.05),减轻结肠组织病理炎症损伤程度,降低血清和结肠组织中促炎性细胞因子IL-6、TNF-α的表达水平(P<0.05,P<0.01),提高抗炎因子IL-10的表达水平(P<0.05,P<0.01)。结论:健脾化滞方对急性UC小鼠有较好的治疗作用,其作用机制可能为修复UC小鼠结肠黏膜局部损伤,调节血清及结肠组织炎症因子水平。

Objective:To study the therapeutic effect of Jianpi Huazhi Formula(JHF)on acute ulcerative colitis(UC)model mice,and explore its mechanism of action based on the expression levels of inflammatory factors in serum and colon tissue of mice.Methods:Seventy-six healthy male C57 mice were randomly divided into normal group,model group,mesalazine group,and low,medium,and high dose JHF groups.An acute UC mouse model was established by drinking 3%dextran sulfate sodium(DSS)solution.The low,medium,and high dose JHF groups were orally administered with 145.8,291.5,583.0 g/L JHF solution respectively,while the mesalazine group was orally administered with 10 g/L mesalazine solution.The symptoms of mice in each group were observed,the disease activity index(DAI)was scored,and the pathological changes of colon tissue were observed by hematoxylin-eosin(HE)staining.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),direct bilirubin(DBil),total bilirubin(TBil),albumin(ALB),alkaline phosphatase(ALP),γ-glutamyl transpeptidase(γ-GT),total bile acid(TBA),creatinine(Cre),uric acid(UA)and blood urea nitrogen(BUN)were measured by rate method and endpoint method.The levels of interleukin(IL)-6,tumor necrosis factor-α(TNF-α)and IL-10 in serum and colon tissue of mice were determined by enzyme-linked immunosorbent assay(ELISA).Results:JHF could effectively improve clinical symptoms such as bloody stools and diarrhea in acute UC mice,improve body mass loss and colon length reduction of mice,reduce DAI score of mice(P<0.05),alleviate the degree of pathological inflammation damage in colon tissue,reduce the expression levels of pro-inflammatory cytokines IL-6 and TNF-α,and increase the expression level of anti-inflammatory factor IL-10(P<0.05,P0.01).Conclusion:JHF has a good therapeutic effect on acute UC mice,and its mechanism of action may be to repair local damage to the colon mucosa of UC mice,regulate the levels of inflammatory factors in serum and colon tissue.