CAJ | 학술논문

目的观察基质细胞衍生因子1(stromal cell-derived factor 1,SDF-1)在小鼠肾脏缺血再灌注损伤(IRI)中表达的动态变化,分析其与巨噬细胞的相关性,探讨其在IRI免疫炎性反应中的作用.方法采用健康雄性C57BL/6小鼠双侧肾蒂夹闭35 min建立肾脏IRI模型.分别于再灌注1d、3d、5d、14 d后收集肾组织样本,HE染色光镜下观察肾脏病理改变;采用实时荧光定量PCR、ELISA法和免疫组织化学法分别检测小鼠肾组织SDF-1的表达及分布.建立小鼠IRI模型前给予腹腔注射氯膦酸二钠脂质体(liposomal clodronate,LC)去除巨噬细胞,免疫荧光染色检测巨噬细胞标志物CD68的表达及分布;检测去除小鼠巨噬细胞后肾脏组织形态学改变及SDF-1的表达变化.结果 HE染色显示IRI小鼠肾脏可见典型肾小管损伤,大量炎性细胞浸润.实时荧光定量PCR和ELISA法结果显示,与假手术组比较,IRI组小鼠肾脏SDF-1mRNA和蛋白表达水平在造模后1d明显升高并达到峰值,差异有统计学意义(均P＜0.05),第5天下降,但仍高于正常水平,14d时恢复至正常水平.免疫组化结果显示,正常小鼠肾脏SDF-1主要表达在皮质,IRI后SDF-1呈高表达并扩展至皮髓交界区域,且SDF-1表达呈时间依赖性下调.与IRI组相比,LC组肾组织CD68表达明显减少,1d时肾组织IR病理改变减轻,且SDF-1表达上调(P＜0.05).结论肾脏IRI初期,SDF-1在肾脏组织中高度表达,并与M1型巨噬细胞有关,临床上也可用作诊断早期急性肾损伤(AKI)的生物学标准.
목적 관찰기질세포연생인자1(stromal cell-derived factor 1,SDF-1)재소서신장결혈재관주손상(IRI)중표체적동태변화,분석기여거서세포적상관성,탐토기재IRI면역염성반응중적작용.방법 채용건강웅성C57BL/6소서쌍측신체협폐35 min건립신장IRI모형.분별우재관주1d、3d、5d、14 d후수집신조직양본,HE염색광경하관찰신장병리개변;채용실시형광정량PCR、ELISA법화면역조직화학법분별검측소서신조직SDF-1적표체급분포.건립소서IRI모형전급여복강주사록련산이납지질체(liposomal clodronate,LC)거제거서세포,면역형광염색검측거서세포표지물CD68적표체급분포;검측거제소서거서세포후신장조직형태학개변급SDF-1적표체변화.결과 HE염색현시IRI소서신장가견전형신소관손상,대량염성세포침윤.실시형광정량PCR화ELISA법결과현시,여가수술조비교,IRI조소서신장SDF-1mRNA화단백표체수평재조모후1d명현승고병체도봉치,차이유통계학의의(균P＜0.05),제5천하강,단잉고우정상수평,14d시회복지정상수평.면역조화결과현시,정상소서신장SDF-1주요표체재피질,IRI후SDF-1정고표체병확전지피수교계구역,차SDF-1표체정시간의뢰성하조.여IRI조상비,LC조신조직CD68표체명현감소,1d시신조직IR병리개변감경,차SDF-1표체상조(P＜0.05).결론 신장IRI초기,SDF-1재신장조직중고도표체,병여M1형거서세포유관,림상상야가용작진단조기급성신손상(AKI)적생물학표준.
Objective To observe the expression of stromal cell-derived factor 1 (SDF-1) in the kidney after ischemic reperfusion injury (IRI),and explore its relationship with macrophage during the IRI kidney.Methods A total of 28 healthy C57BL/6 male mice were used to establish renal IRI model by clamping both pedicles for 35 min followed by reperfusion.Kidney tissue samples were collected at indicated time points.Renal histological changes were estimated.The expression of SDF-1 was determined by immunohistochemistry,ELISA and real-time PCR.After the liposomal clodronate was injected intraperitoneally,the location of CD68 was observed by immunofluorescence.Renal histology and protein expression of SDF-1 were also detected.Results Compared with sham-operated group,classical tubular damage was found in IRI group,accompanied by a large number of inflammatory cells.The expression of total renal SDF-1 peaked on day 1 and decreased to control levels in the following days.SDF-1 in healthy kidney was localized at cortex,but spread to the corticomedullary area of the kidney during IRI.Compared with IRI groups,elimination of macrophage by injection of liposomal clodronate alleviated renal IRI and down-regulated the expressions of CD68 while up-regulating SDF-1.Conclusions SDF-1 expression is up-regulated in IRI kidney and is associated with macrophage.SDF-1 may play a role in the early phase of acute kidney injury and it may be a new marker in diagnosis of AKI.