摘要
目的讨论小鼠肾小管上皮细胞(mRTEC)来源外泌体(mRTEC-EXO)对脓毒症肾脏损伤炎症反应的影响。方法制备mRTEC-EXO,使用透射电子显微镜观察其形态,通过共聚焦显微镜观测其是否被胚胎肾(EK)细胞摄取。将84只SPF小鼠随机分为对照组[腹腔注射生理盐水(NS)]、LPS组(腹腔注射LPS)、LPS+mRTEC-EXO组(腹腔注射LPS同时尾静脉注射mRTEC-EXO),每组各28只。NS注射后或LPS干预12 h后,随机处死各组小鼠均12只,收集新鲜肾脏组织切片后行HE及免疫组化染色,观察肾小管、肾小球形态变化及肾小管间质炎性细胞、中性粒细胞的浸润程度。每组剩余16只小鼠继续观察至150 h,记录其存活情况。采用Western blot检测mRTEC-EXO中CD9、CD63、Calnexin及3组小鼠肾脏组织中MPO、MDA、TNF-α和IL-1β的表达水平。结果Western blot结果显示,mRTEC-EXO表面标志物CD9和CD63呈高表达,而阴性标志物Calnexin呈低表达。共聚焦显微镜观察结果显示,EK细胞可主动摄取mRTEC-EXO。对照组小鼠生存率均高于LPS组和LPS+mRTEC-EXO组,LPS+mRTEC-EXO组生存率高于LPS组(P<0.05)。与对照组比较,LPS组和LPS+mRTEC-EXO组小鼠肾脏组织中TNF-α、IL-1β、MDA、MPO表达水平均明显升高;与LPS组比较,LPS+mRTEC-EXO组TNF-α、L-1β、MDA、MPO表达水平均明显降低(P<0.05)。与对照组比较,LPS组及LPS+mRTEC-EXO组小鼠肾脏组织炎性浸润程度明显增加。与LPS组比较,LPS+mRTEC-EXO组小鼠肾脏组织炎性浸润程度明显降低。结论mRTEC-EXO可抑制中性粒细胞浸润并减弱脓毒症肾脏损伤炎症反应。
Objective To investigate the effect of exosomes derived from mouse renal tubular epithelial cells(mRTEC-EXO)on inflammation response septic kidney injury.Methods mRTEC-EXO was prepared,the morphology of mRTEC-EXO was observed with transmission electron microscopy and its uptake by embryonic kidney(EK)cells with confocal microscopy.Eighty-four SPF mice were randomly divided into control group[normal saline(NS)was injected intraperitoneally],LPS group(LPS was injected intraperitoneally)and LPS+mRTEC-EXO group[LPS was injected intraperitoneally and mRTEC-EXO was injected through the tail vein],with 28 mice in each group.After NS intervention or 12 h after LPS intervention,12 mice in each group were randomly sacrificed.Fresh kidney tissue sections were collected and stained with HE and immunohistochemistry to observe the morphological changes of renal tubules and glomeruli,the infiltration degree of inflammatory cells and neutrophils in renal tubule interstitium.The remaining 16 mice in each group continue to observe 150 h,record its survival situation.The expression levels of CD9,CD63 and Calnexin in mRTEC-EXO and MPO,MDA,TNF-αand IL-1βin kidney tissue of the 3 groups were detected by Western blotting.Results Western blotting results showed that the surface markers CD9 and CD63 of mRTEC-EXO were highly expressed,while the surface marker Calnexin was lowly expressed.The results of confocal microscopy showed that EK cells could actively take up mRTEC-EXO.The survival rate of mice in control group was higher than that in LPS group and LPS+mRTEC-EXO group,and the survival rate of mice in LPS+mRTEC-EXO group was higher than that in LPS group(P<0.05).Compared with the control group,the expression levels of TNF-α,IL-1β,MDA and MPO in kidney tissues of mice in LPS group and LPS+mRTEC-EXO group were significantly increased;Compared with LPS group,the expression levels of TNF-α,L-1β,MDA and MPO in LPS+mRTEC-EXO group were significantly decreased(P<0.05).Compared with the control group,the characteristics of inflammatory infiltration were significantly increased in LPS group and LPS+mRTEC-EXO group;Compared with LPS group,LPS+mRTEC-EXO group showed significantly lower inflammatory infiltration characteristics.Conclusion mRTEC-EXO inhibite neutrophils infiltration and reduce the inflammatory response to septic renal injury.
作者
徐志育
朱永
田佳
李娜
宋海
Xu Zhiyu;Zhu Yong;Tian Jia;Li Na;Song Hai(Department of Critical Care Medicine,Hainan Provincial People’s Hospital,Haikou 570311,China;不详)
出处
《临床内科杂志》
CAS
2022年第9期625-629,共5页
Journal of Clinical Internal Medicine
基金
海南省卫生健康行业科研项目(19A200005)
关键词
外泌体
脓毒症
中性粒细胞
浸润
肾损伤
炎症反应
Exosomes
Sepsis
Neutrophils
Infiltration
Renal injury
Inflammatory response
