中药排气汤对术后肠梗阻小鼠胃肠功能的作用及机制研究

Effect and mechanism of Chinese medicine Paiqi Decoction on gastrointestinal function of mice with postoperative ileus

目的:研究中药排气汤对术后肠梗阻小鼠胃肠功能的作用及机制。方法:取7周龄SPF级C57BL/6J小鼠24只,随机分为假手术组、模型组、莫沙必利组、排气汤组,每组6只,采用经典小肠干扰术诱导小鼠术后肠梗阻模型。手术24 h后观测各组小鼠胃肠运动功能、肠道组织结构,ELISA试剂盒检测小鼠血清促炎因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和抗炎因子白细胞介素-10(IL-10)的水平,免疫组织化学法检测小鼠小肠Cajal间质细胞(interstitial cells of Cajal,ICC)及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的表达情况,Western Blot检测小鼠小肠中Toll样受体4(TLR4)、活化丝裂原激活的蛋白激酶p38(p38MAPK)和核转录因子-κB p65(NF-κB p65)表达水平。结果:与假手术组相比,模型组小鼠小肠推进率显著下降(P<0.01),血清炎症因子含量明显升高(P<0.05);与模型组相比,排气汤组小鼠小肠推进率明显升高(P<0.05),血清中炎症因子含量明显降低(P<0.05)。Western Blot结果显示,排气汤组能显著降低术后肠梗阻小鼠小肠TLR4、p38MAPK和NF-κB p65蛋白表达(P<0.05)。结论:中药排气汤可以改善术后肠梗阻小鼠胃肠道功能,其作用机制与恢复ICC的功能,抑制TLR4、p38MAPK和NF-κB p65的表达,减轻炎症反应有关。

OBJECTIVE To study the effect and mechanism of Chinese medicine Paiqi Decoction on gastrointestinal function of mice with postoperative ileus.METHODS Twenty-four seven-week-old SPF C57 BL/6 J mice were randomly divided into sham operation group,model group,moxapbide group and Paiqi Decoction group,with 6 mice in each group.Intestinal obstruction model was induced by classical intestinal interference.Gastrointestinal motor function and intestinal tissue structure of mice in each group were observed 24 h after operation.Serum levels of pro-inflammatory cytokine interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and anti-inflammatory cytokine interleukin-10(IL-10)were detected by ELISA kit.The expression of Interstitial cells of Cajal(ICC)and inducible nitric oxide synthase(iNOS)in small intestine of mice were detected by immunohistochemistry.The expression levels of toll-like receptor 4(TLR4),activated mitogen-activated protein kinase p38(p38 MAPK)and nuclear transcription factor-κB p65(NF-κB p65)in small intestine were detected by Western Blot.RESULTS Compared with sham operation group,small intestine propulsion rate of model group was significantly decreased(P<0.01),and serum inflammatory factor content was significantly increased(P<0.05).Compared with model group,intestinal propulsion rate of Paiqi Decoction group was significantly increased(P<0.05),and the content of inflammatory factors in serum was significantly decreased(P<0.05).Western Blot results showed that the expression of TLR4,p38 MAPK and NF-κB p65 protein in small intestine of mice with postoperative ileus was significantly decreased in Paiqi Decoction group(P<0.05).CONCLUSION Chinese medicine Paiqi Decoction can improve gastrointestinal function of mice with postoperative ileus,and its mechanism is related to restoring ICC function,inhibiting the expression of TLR4,p38 MAPK and NF-κB p65,and alleviating inflammation.