芍药汤调节湿热型溃疡性结肠炎大鼠JAK2/STAT3和SOCS3的分子机制

Effect of Shaoyaotang in Adjusting Molecular Mechanism of JAK2/STAT3 and SOCS3 in Syndrome Ulcerative Colitis Rats

目的:观察芍药汤通过治疗湿热型溃疡性结肠炎(ulcerative colitis,UC)大鼠模型,对结肠组织JAK2/STAT3和SOCS3的影响,从分子水平上探讨芍药汤治疗湿热型UC的机制。方法:Wistar大鼠雌雄各60只,分为空白组、模型组、芍药汤高、中、低(24,12,6 g·kg^(-1))剂量组、阳性药组(柳氮磺砒啶组,1 g·kg^(-1)),通过高脂高糖辛辣食物加免疫复合法,2,4,6-三硝基苯磺酸(TNBS)结合乙醇复合法复制湿热型UC大鼠模型,芍药汤高、中、低剂量灌服,柳氮磺砒啶组予柳氮磺砒啶研磨成粉灌服,空白组及模型组予等体积生理盐水灌胃,连续21 d。采集结肠组织,运用苏木素-伊红(HE)染色观察病理切片、实时荧光定量PCR法(Real-time PCR)检测基因含量、蛋白免疫印迹法(Western blot)检测蛋白含量。结果:模型组产生炎症反应,镜下出现黏膜损伤、溃疡,提示造模成功。与空白组比较,模型组JAK2,STAT3蛋白及基因表达明显升高(P<0.05),SOCS3明显下降(P<0.05);与模型组比较,各治疗组JAK2,STAT3蛋白及基因表达明显下降,其中芍药汤高剂量组最为显著(P<0.01),SOCS3明显升高,芍药汤高剂量组最为显著(P<0.05)。结论:芍药汤能够改善湿热型UC大鼠结肠黏膜组织的病变程度,有效减轻炎症反应,与SOCS3抑制JAK2/STAT3信号通路,产生负反馈调节,降低其活化有关。

Objective： To observe the effect of Shaoyaotang on colon tissues JAK2/STAT3 and SOCS3, in order to discuss the mechanism of Shaoyaotang by treating the rat model of syndrome type ulcerative colitis （UC）. Method： A total of 120 Wistar rats, including 60 male rats and 60 female rats, were divided into 6 groups： blank group, model group, high-dose （24 g·kg^-1）, medium-dose （12 g·kg^-1）, low-dose （6 g·kg^-1） Shaoyaotang groups, and western medicine group （sulfasalazine group, 1 g·kg^-1）. High-fat, high-sugar, spicy food plus immune complex, TBNS （2, 4, 6-trinitro-benzene-sulfonic acid） and ethanol were used to replicate the syndrome type UC rat model. Wistar rats in different groups were given different treatments for consecutively 21 days. In Shaoyaotang group, Wistar rats were given high, medium and low-dose Shaoyaotang by garage. In western medicine group, they were administered with sulfasalazine powder. In blank group and model group, they were given isopyknic normal saline for intragastric administration. After collecting abdominal aorta serum and colon tissues, sue-eosin methyl （HE） staining was used to observe the pathological section; Real-time PCR method was used to detect the contents of genes. And Western blot method was used to detect the contents of protein. Result： The model group showed inflammatory reaction, mucosa damage and ulceration under the endoscope, which indicated the successful modeling. Compared with blank group, model group＇s JAK2 and STAT3 protein and gene expressions increased significantly （P 〈 0.05） , while SOCS3 decreased significantly （P 〈 0.05 ）. Compared with the model group, in Shaoyaotang treatment groups, JAK2 and STAT3 protein and gene expressions were significantly decreased, and high-dose Shaoyaotang group showed the most obvious effect （P 〈 0.01 ） ; SOCS3 increased dramatically, and high-dose Shaoyaotang group showed the most obvious effect （P 〈 0.05 ）. Conclusion： Shaoyaotang can improve the pathological changes in colon mucosal tissues of rats with syndrome type UC and reduce inflammation, which may be related to the inhibition of JAK/STAT3 signaling pathway by SOCS3, the generation of negative feedback regulation and the decrease in activation.