雷公藤多苷对溃疡性结肠炎大鼠炎症因子及结肠组织丝裂原p38活化蛋白激酶和核因子-κBp65蛋白表达的影响

Effect of glucosidorum tripterygii tororum on the expression of inflammatory factor and p38MAPK,nuclear factor-κBp65 protein in ulcerative colitis rats

目的研究雷公藤多苷对溃疡性结肠炎大鼠炎性因子及结肠组织丝裂原p38活化蛋白激酶(p38MAPK)和核因子-κBp65(NF-κBp65)蛋白表达的影响。方法从32只Wistar大鼠中随机选取8只为正常组不做任何处理,24只构建溃疡性结肠炎大鼠模型,死亡2只,将剩余22只模型大鼠随机分为模型组(n=7)、实验组(n=8)和阳性对照组(n=7)。正常组和模型组均灌胃生理盐水10 m L·kg^(-1),试验组灌胃雷公藤多苷20 mg·kg-1,阳性对照组灌胃柳氮磺砒啶片0.5 mg·kg^(-1),连续灌服14 d。用酶联免疫吸附实验测定各组大鼠血清炎症因子水平,用苏木精-伊红染色法观察各组大鼠结肠组织病理变化,用蛋白质免疫印迹法检测结肠组织p38MAPK、NF-κBp65蛋白的表达。结果模型组大鼠血清IL-4水平低于正常组,实验组和阳性对照组IL-4水平高于模型组(P<0.01);模型组大鼠血清IL-6水平高于正常组、实验组和阳性对照组(P<0.01);模型组、实验组和阳性对照组大鼠血清IL-1β水平高于正常对照组,实验组和阳性对照组IL-1β水平低于模型组(P<0.01)。模型组、实验组和阳性对照组结肠组织损伤评分分别为(4.56±0.88),(1.65±0.53),(1.50±0.54)分,明显高于对照组的(0.23±0.32)分(P<0.01);与模型组比较,实验组和阳性对照组结肠组织损伤评分均降低(均P<0.01)。模型组p38MAPK和NF-κBp65蛋白相对表达量分别为1.62±2.39,1.53±3.21,明显高于正常对照组的0.74±0.11,0.63±0.09;实验组和阳性对照组p38MAPK蛋白相对表达量分别为0.93±0.16,0.78±0.19,NF-κBp65蛋白相对表达量分别为0.81±0.23,0.72±0.16,均显著低于模型组(均P<0.01)。结论雷公藤多苷可有效降低溃疡性结肠炎大鼠血清炎症因子水平,改善结肠黏膜组织形态,其分子实质可能与降低p38MAPK及NF-κBp65蛋白表达量,调控其所介导的炎症反应通路有关。

Objective To observe the effect of glucosidorum tripterygii tororum on the expression of inflammatory factor and p38 mitogen-activated protein kinases（ p38 MAPK）,nuclear factor-κB（ NF-κB） p65 protein in ulcerative colitis rats. Methods Eight Wistar rats were randomly selected as normal group,without any treatment,24 rats were modeled on ulcerative colitis rats,2 rats dead,the remaining 22 rats were randomly divided into model group（ n = 7）,test group（ n = 8） and positive control group（ n = 7）. The normal group and model group weregavaged with 0. 9% Na Cl 10 m L·kg-（-1）,test group was gavaged with glucosidorum tripterygii tororum 20 mg·kg-（-1）,positive control group was gavaged with sulfasalazine tablets 0. 5 mg·kg-1,all rats were continuously gavaged for 14 d. The levels of serum inflammatory cytokines were measured by enzyme-linked immunosorbent assay,the histopathological changes of colon were observed by hematoxylin-eosin staining,the expression of p38 MAPK and NF-κBp65 protein were detected by Western blotting. Results The serum IL-4 level in model group was significantly lower than that in normal group,IL-4 levels in test and positive control group were higher than those in model group（ P〈0. 01）.The level of serum IL-6 in model group was higher than those in normal group,test group and positive control group（ P〈0. 01）. The levels of serum IL-1β in model group,test group and positive control group were higher than that in normal group（ P〈0. 01）,the levels of IL-1β in test group and positive control group were lower than that in model group（ P〈0. 01）. The colon tissue injury score of model group,test group and positive control group were4. 56 ± 0. 88,1. 65 ± 0. 53,1. 50 ± 0. 54,higher than that in normal group,which was 0. 23 ± 0. 32（ P〈0. 01）,compared with model group,the scores of colonic tissue injury in test group and positive control group decreased（ P〈0. 01）. The relative expression of p38 MAPK and NF-κBp65 in model group were 1. 62 ± 2. 39,1. 53 ± 3. 21,higher than those in normal group,wihch were 0. 74 ± 0. 11,0. 63 ± 0. 09. The relative expression levels of p38 MAPK protein in test group and positive control group were 0. 93 ± 0. 16,0. 78 ± 0. 19,the relative expression levels of NF-κBp65 protein were 0. 81 ± 0. 23,0. 72 ± 0. 16,lower than model group（ all P〈0. 01）. Conclusion Glucosidorum tripterygii tororum can effectively reduce the levels of serum inflammatory cytokines in ulcerative colitis rats and improve the morphology of colonic mucosa. The molecular substance may be related to the decrease of p38 MAPK and NF-κBp65 protein expression and the regulation of inflammatory pathway.