摘要
目的观察熊果酸(UA)对肝纤维化模型大鼠肝内胶原沉积的改善作用及其对NADPH氧化酶2(NOX2)/活性氧簇(ROS)/NLRP3炎性小体活化的影响。方法将大鼠随机分为对照组、CCl4模型组及UA治疗组。用CCl4诱导构建SD大鼠肝纤维化模型,一半用作UA治疗组。用试剂盒检测血清ALT含量;HE染色观察肝脏病理;天狼星红染色观察肝内胶原沉积;RT-q PCR法检测Nox2、Nlrp3、caspase1及IL-1βmRNA表达量;Western blot及免疫组化检测肝脏中NOX2、NLRP3、casepase-1 p45、caspase-1 p10及IL-1β蛋白表达;DCFH-DA荧光探针法检测肝脏组织内ROS水平。结果与对照组相比,CCl4模型组血清ALT水平升高(P<0.05),Ishak's肝纤维化评分明显上升,胶原沉积明显增多(P<0.05),Nox2、Nlrp3、Caspase1及IL-1βmRNA的表达水平明显上升(P<0.05);NOX2、NLRP3、caspase-1 p10及IL-1β蛋白的表达均出现明显增加,肝组织内ROS含量增加(P<0.05);与CCl4模型组相比,UA治疗组血清ALT含量下降(P<0.05),肝脏Ishak's肝纤维化评分及胶原沉积减少(P<0.05),Nox2、Nlrp3、caspase1及IL-1βmRNA的表达水平明显下降(P<0.05),NOX2、NLRP3、caspase-1 p10及IL-1β蛋白表达明显下降,肝脏组织内ROS水平显著改善(P<0.05)。结论 UA减轻肝脏炎性反应并减少肝内胶原沉积,其机制可能与其抑制肝纤维化大鼠肝内NOX2/ROS/NLRP3炎性小体活化及IL-1β的释放减少有关。
Objective To investigate the effect of ursolic acid(UA)on NOX2/ROS/NLRP3 inflammasome activation in carbon tetrachloride(CCl 4)-induced liver fibrosis SD rat,and to observe the improvement of collagen deposition in liver tissues.Methods All rats were randomly divided into 3 group:control group,CCl 4 model group,UA treatment group.Liver fibrosis model SD rats was established by the CCl 4-induced method and half of them was used as UA treatment group.Serum ALT was detected by ALT detection kit.The liver pathology and collagen deposition were observed by HE and Sirius-red staining.The mRNA expression of Nox2,Nlrp3,Caspase1,IL-1βin liver tissues was detected by RT-qPCR.The protein expression of NOX2,NLRP3,caspase-1 and IL-1βin liver tissues was detected by Western blot and immunohistochemistry and the ROS generation in liver tissues was detected by DCFH-DA fluorescence probe.Results Compared with control group,in the CCl 4 model group,the serum ALT was much higher(P<0.05);the Ishak s fibrosis score and collagen deposition was significantly increased(P<0.05)and mRNA of Nox2,Nlrp3,Caspase1,IL-1βwas increased.In addition,both the NOX2,NLRP3,caspase-1 p10 and IL-1βprotein expression and ROS level(P<0.05)of CCl 4 model group were significant increased.Compared with CCl 4 model group,in the UA treatment group Ishak s fibrosis score,collagen deposition and ALT decreased.Both mRNA expression of the Nox2,Nlrp3,Caspase1,IL-1βand protein expression of NOX2,NLRP3,caspase-1 p10 and IL-1βas well as ROS were significant decreased,but the caspase-1 p45 protein level has no difference among all these groups(P>0.05).Conclusions Ursolic acid attenuates the liver injury and reduces the collagen deposition,which may relate to its inhibitory effects on NOX2/ROS/NLRP3 inflammasome activation to reduce IL-1βreleasing.
作者
黄晨恺
甘达凯
张望
罗方云
陈江
汪安江
李弼民
朱萱
HUANG Chenkai;GAN Dakai;ZHANG Wang;LUO Fangyun;CHEN Jiang;WANG Anjiang;LI Bimin;ZHU Xuan(Dept.of Digestive Diseases,the First Affiliated Hospital,Nanchang University;Jiangxi Provincial Institute of Gastroenterology,Nanchang 330006,China)
出处
《基础医学与临床》
CSCD
2018年第4期485-491,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(81660110)
