清瘟败毒饮对脓毒症急性肺损伤大鼠JAK2/STAT3信号通路及其负反馈机制的影响

The effect and mechanism of Qingwen Baidu Decoction on JAK2/STAT3 pathway and negative feedback in sepsis rats

目的:观察清瘟败毒饮对脓毒症急性肺损伤(ALI)大鼠JAK2/STAT3信号通路及其负反馈机制的影响。方法:将50只雄性Wistar大鼠按照体重随机分5组,即空白对照组、ALI模型组、地塞米松0. 27mg/kg组、清瘟败毒饮7. 5和15 g/kg组。除空白对照组尾静脉注射等体积生理盐水外,其余各组均通过尾静脉注射内毒素3mg/kg复制脓毒症急性肺损伤大鼠模型;清瘟败毒饮给药组和地塞米松组分别于造模前预先灌胃给药,每天1次,连续6天,于末次给药30分钟后尾静脉注射内毒素造模;造模24小时后,酶联免疫吸附法(ELISA)测定大鼠肺组织中TNF-α、IL-1β和IL-6表达水平;电镜观察大鼠肺组织肺泡Ⅱ型细胞的微观病理变化; Westernblot测定各组大鼠肺组织中p-JAK2、p-STAT3、SOCS1和PIAS3蛋白的表达水平。结果:模型组肺组织中TNF-α、IL-1β、IL-6、p-JAK2、p-STAT3、SOCS1和PIAS3蛋白表达水平显著高于空白对照组,肺泡Ⅱ型细胞形态不规则,细胞表面微绒毛显著减少,胞核变形,染色质边集,板层小体空泡化并排空显著增多,线粒体、高尔基体和内质网等细胞器明显肿胀。与模型组相比,清瘟败毒饮给药15 g/kg和7. 5g/kg组肺泡Ⅱ型细胞微观病理损伤明显减轻,肺组织中TNF-α、IL-1β、IL-6、p-JAK2、p-STAT3和PIAS3蛋白表达水平显著降低,SOCS1负反馈调节蛋白表达显著升高。结论:清瘟败毒饮可通过有效抑制脓毒症ALI大鼠肺组织炎性因子(TNF-α、IL-1β、IL-6)及其介导的JAK2/STAT3信号通路,提高该通路负反馈调节机制,对ALI模型大鼠起到保护作用。

Objective： Through the observation of LPS induced by sepsis induced acute lung injury（ALI） expression in pathological model and JAK2/STAT3 pathway protein in lung tissue of rats,to explore the Qingwen Baidu Decoction on sepsis induced acute lung injury（ALI） in treatment effect of intervention and mechanism in rats. Methods： From 50 male healthy adult Wistar,Were randomly divided into five groups,such as the normal control group（normal saline Ig）,ALI model group（saline,Ig） Qingwen Baidu Decoction high dose group（Ig15 g crude drug/kg）,Qingwen Baidu Decoction low dose group（Ig 7. 5 g crude drug/kg）,positive control group（dexamethasone0. 27 mg/kg Ig） except the blank group were by cecal ligation of sepsis induced acute lung injury rat model. Through the EVLW,Pa CO2,Pa O2 to be observed,The structural change of ultraminiature lung tissues to determine the degree of inflammatory reaction by transmission electron microscope（TEM）,rats in each group were determined the expression of p-JAK2,p-STAT3,SOCS1 and PIAS3 proteins of lung tissue modeling using Western blotting and the expression of TNF-α、IL-1β and IL-6 by ELISA after 24 hours. Results： Compared to the model group,EVLW,Pa CO2,pathological TypeⅡ alveolar cell injury of observation data and expression of TNF-α,IL-1β,IL-6,p-JAK2,p-STAT3 and PIAS3 protein in lung content in Qingwen Baidu Decoction groups（7. 5 - 15 g/kg） significantly reduced（P 〈 0. 05 或 P 〈 0. 01）,In addition the expression of SOCS1 and Pa O2 in content in Qingwen Baidu Decoction groups（7. 5 - 15 g/kg） and Dexamethasone group（0. 27 mg/kg） significantly increased（P 〈 0. 05）. ALI model group had shows the typeⅡ alveolar cells became small,and cell superficial microvilli disappeared,Osmiophilic lamellar body became atrophy,lacked lamellar structure and formed a solid globule with high electrom dense. Conclusion： The Qingwen Baidu Decoction could effectively improve the pathological and histological changes in Type Ⅱalveolar cell and inhibit expression of TNF-α,IL-1β,IL-6 of the lung in ALI rats,the therapeutic mechanism might be related to the expressions of the JAK2/STAT3 pathway and negative feedback.