活血健脾补肾法对结肠炎小鼠结肠组织TNF-α及其mRNA表达的影响

Effects of Huoxue, Jianpi and Bushen recipe on expression of TNF-α and its mRNA in mice with colitis

目的:观察TNF-α及其mRNA在结肠炎小鼠结肠组织的表达,探讨活血健脾补肾法方药对其影响．方法:用右旋葡聚糖硫酸钠(DSS)诱导小鼠产生溃疡性结肠炎模型．将小鼠随机分为空白对照、模型、柳氮磺胺吡啶(SASP)、活血法、健脾法及补肾法组;免疫组化、原位杂交法分别观察治疗后小鼠结肠组织TNF-α及其mRNA的表达．结果:模型组小鼠结肠组织学损伤积分(炎症、病度深度、隐窝破坏)分别为4．85±2．1,5．77±2．2,7．76±2．4;活血法组 (2．24±2．4,2．53±2．5,3．49±2．3)、健脾法组(2．76+2．2,2．89±2．4, 3．87±2．3)、补肾法组(2．12±2．3,2．33±2．2,3．44±2．4)与模型组比较有显著意义(4．85±2．1,5．77±2．2,7．76±2．4)(均 P<0．05)．模型组小鼠结肠组织TNF-α及其mRNA的表达(染色积分、平均光密度、平均灰度)分别是6．8±1．4,0．35±0．03． 78．6±4．4;活血法组(3．7±1．1,0．18±0．05,137．9±6．7)、健脾法组(3．4±1．3,0．16±0．03,155．1±8．8)、补肾法组(3．1± 1．5,0．17±0．04,145．6．2±7．6)与模型组比较有显著意义(6．8± 1．4,0．35±0．03,78．6±4．4)(均P<0．05)．结论:TNF-α参与DSS诱导小鼠溃疡性结肠炎的形成,活血、健脾及补肾法方药通过下调TNF-α的表达发挥防治作用．

AIM： To observe the expression of tumor necrosis factor-α （TNF-α） and its mRNA in colitis mice, and to explore the effects of Huoxue JianpiBushen recipe on it. METHODS： The mouse model of ulcerative colitis was induced by dextran sulfate sodium （DSS）. The mice were randomly divided into six groups： normal control, model, sulfasalazine （SASP）, Huoxun, Jianpi, and Bushen group. The expression of TNF-α and its mRNA in the mice were examined by immunohistochemistry and in situ hybridization. RESULTS： The colonic histological scores （for inflammation, depth of the illness, damage of the recess） of the mice in the model group were 4.85±2.1, 5.77±2.2, and 7.76±2.4, respectively, which were significantly higher than those in Huoxue group （2.24±2.4, 2.53± 2.5, 3.49±2.3）, Jianpi group （2.76±2.2, 2.89±2.4, 3.87± 2.3）, and Bushen group （2.12±2.3, 2.33±2.2, 3.44±2.4） （all P 〈0.05）. The expression of TNF-α （integral after staining, mean optical density, mean grey） in model group （6.8±1.4, 0.35±0.03, 78.6±4.4） was significantly different from those in Huoxue group （3.7±1.1, 0.18± 0.05, 137.9±6.7）, Jianpi group （3.4±1.3, 0.16±0.03, 155.1±8.8）, and Bushen group （3.1±1.5, 0.17±0.04 145.62±7.6）（all P 〈0.05）. CONCLUSION： TNF-α plays an important role in the occurrence of ulcerative colitis induced by DSS, and the recipe of Huoxue, Jianpi, and Bushen can prevent and cure this disease by down-regulating the expression of TNF-α.