连理汤对TNBS诱导大鼠溃疡性结肠炎模型结肠及外周血中炎症相关因子影响的拆方研究

Decomposition Study on Effects of Lianli Tang on Colon and Peripheral Blood Inflammatory Factors in TNBS-induced Rat Model

摘要:

目的：采用2，4，6-三硝基苯磺酸（TNBS）诱导的大鼠溃疡性结肠炎（UC）模型，观察连理汤各拆方药组及其配伍对UC大鼠外周血及结肠组织中细胞因子等含量的影响。方法：将连理汤拆分成补脾益肠药（A因素）、清热燥湿药（B因素）、温中散寒药（C因素），以含和不含该因素为1，2水平，采用正交设计表L₈（2⁷）设试验1~7组和UC组，增设柳氮磺吡啶组（SASP）与正常对照组。采用Sprague Dawley大鼠，前9组按TNBS乙醇灌肠法塑造UC模型，试验1~7组分别灌胃相应的拆方浸膏混悬液，剂量依次为1.02，1.47，0.98，1.24，0.29，0.145，0.087 g·kg^-1，SASP组灌服柳氮磺吡啶（0.4 g·kg^-1），以UC大鼠外周血及结肠组织中细胞因子等含量作为考察指标，观察灌服连理汤拆方14 d后各组对其含量的影响。结果：UC组大鼠结肠炎症环氧合酶-2（Cox-2）、白介素-1α（IL-1α）含量较正常对照组有显著升高（P<0.01），金属蛋白酶类组织抑制剂（tissue inhibitor of metalloproteinases 1，TIMP-1）和肿瘤坏死因子-α（tumor necrosis factor alpha，TNF-α）表达也有明显增强。补脾益肠药（A）在抑制结肠Cox-2方面影响显著（P<0.05）；各因素对结肠中IL-1α含量的影响均不显著，但全方效果显著（P<0.01）；补脾益肠药和清热燥湿药配伍（即A×B交互作用）对降低结肠TNF-α含量影响显著（P<0.05）；温中散寒药（C）以及补脾益肠药和温中散寒药配伍（即A×C交互作用）对抑制UC大鼠结肠组织中TIMP-1 含量有极显著影响（P<0.01），清热燥湿药（B）以及清热燥湿药和温中散寒药配伍（即B×C交互作用）对该指标也有显著影响（P<0.05）。UC大鼠血清中IL-1α，TNF-α明显下降，拆方各因素均无显著作用；UC大鼠血清TIMP-1明显升高，温中散寒药（C）对抑制UC模型大鼠外周血中TIMP 含量有显著性影响（P<0.05）。结论：补脾益肠药在连理汤的整体疗效中起主导作用，温中散寒药及其与温补药配伍也起重要作用。

Abstract:

Objective: To carry out decomposition study on effects of Lianli Tang on colon and peripheral blood inflammatory factors in TNBS-induced rat model. Method: Lianli Tang were divided into three groups, tonify the spleen drugs (Panax Ginseng, Poria cocos, Atractylodes macrocephala koidz, Glycyrrhizae Radix et Rhizoma; symbol as A), Warming the middle and dissipating cold drugs (Zingiberis Rhizoma;symbol as B), clearing heat and drying dampness drugs (Coptidis Rhizoma; symbol as C), as three factors to design the experiment using orthogonal design table L₈ (2⁷), set up 1-7 group and UC group, then add sulfasalazine (SASP) and normal controls group (Control). Sprague Dawley rats were used to duplicate UC model by TNBS/ethanol solution through enema, 1-7 group was administrated respectively by Lianli Tang decompositions through intragastric treatment, which the corresponding doses is 1.02, 1.47, 0.98, 1.24, 0.29, 0.145, 0.087 g·kg^-1, SASP group rats were fed by sulfasalazine (0.4 g·kg^-1). Cytokines of peripheral blood and colon were observed after rats were treated for 14 days. Result: Cyclooxygenase-2 (Cox-2), interleukin-1α ( IL-1α) of UC rats were increased significantly than control (P<0.01), tumor necrosis factor-α (TNF-α) expression were also significantly enhanced, A of Liali Tang significantly affect the effect of inhibiting colon Cox-2 (P<0.05), each factors except the whole formula, have no effect in regulating colonic IL-1α, A×B significantly affected the results (P<0.05) in reducing colonic TNF-α, C, A×C (P<0.01) and B, B×C (P<0.05) in inhibiting colon tissue inhibitor of metall oproteimses 1 (TIMP-1) levels. Serum IL-1α, TNF-α of UC rats decreased apparently, each decomposition factor had no effect on the result;serum TIMP-1 was obviously increased, decomposition C show effect on inhibiting TIMP-1 (P<0.05). Conclusion: Tonify the spleen drugs played a major role in the overall effect of Lianli Tang, Warm the middle and dissipate cold drugs and its compatibility with tonify the spleen drugs also play an important role.