摘要
目的:探究膜联蛋白A1(ANXA1)拟肽Ac2-26对脓毒症大鼠急性肾损伤(AKI)及中性粒细胞凋亡的影响。方法:实验分组包括对照组、Ac2-26组、AKI组、AKI+Ac2-26组,每组15只大鼠。采用盲肠结扎穿孔术建立脓毒症引发AKI模型后,静脉输注Ac2-26进行治疗,1次/d,持续14 d;结束后,ELISA检测各组大鼠血清肌酐(Scr)、尿素氮(BUN)、IL-1β、IL-6及TNF-α水平,HE染色和过碘酸雪夫氏(PAS)染色观察各组大鼠肾组织病理学变化,免疫组织化学染色检测各组大鼠肾组织内ANXA1表达情况;从大鼠外周血中分离中性粒细胞,吉姆萨染色和台盼蓝染色检测细胞纯度与活力;Annexin V-FITC/PI双染法和TUNEL染色测定各组大鼠中性粒细胞凋亡水平。结果:与对照组比较,AKI组大鼠血清中Scr与BUN水平升高(P<0.05),IL-1β、IL-6及TNF-α水平也升高(P<0.05),肾组织内肾小管和肾小球均发生明显损伤,并伴随大量炎症细胞浸润,病理学评分升高(P<0.05),ANXA1阳性染色面积比例减小(P<0.05);经吉姆萨染色和台盼蓝染色鉴定分离到的中性粒细胞形态完整,活性较高;与对照组比较,AKI组大鼠中性粒细胞凋亡率下降(P<0.05),TUNEL阳性率降低(P<0.05)。与AKI组比较,AKI+Ac2-26组血清中Scr与BUN水平降低(P<0.05),IL-1β、IL-6及TNF-α水平也均降低(P<0.05),肾组织内肾小管与肾小球的病理表现明显减轻,病理学评分降低(P<0.05),而ANXA1阳性染色面积比例增大(P<0.05),同时,大鼠中性粒细胞凋亡率升高(P<0.05),TUNEL阳性率也升高(P<0.05)。结论:ANXA1拟肽Ac2-26能够提高脓毒症大鼠AKI中肾组织内ANXA1表达,促进中性粒细胞凋亡,对脓毒症引发的大鼠肾组织损伤具有保护作用。
Objective:To explore the effect of Annexin A1(ANXA1)peptidomimetic Ac2-26 on acute kidney injury(AKI)and neutrophil apoptosis in septic rats.Methods:Experimental groups included control group,Ac2-26 group,AKI group,AKI+Ac2-26 group,with 15 rats in each group.After cecal ligation and perforation were used to establish a sepsis-induced AKI model,Ac2-26 was intravenously infused for treatment,once a day for 14 days;after the end,ELISA was used to detect levels of serum creatinine(Scr),urea nitrogen(BUN),IL-1β,IL-6 and TNF-α;HE staining and periodic acid Schiff(PAS)staining were used to observe the pathological changes of rat kidney tissues in each group;immunohistochemical staining was used to detect expression of ANXA1 in kidney tissue of each group of rats;neutrophils were isolated from rat peripheral blood,Giemsa staining and trypan blue staining were used to detect cell purity and viability;Annexin V-FITC/PI double staining method and TUNEL staining were used to determine apop-tosis level of neutrophils in each group.Results:Compared with control group,levels of Scr and BUN in serum of rats in AKI group were increased(P<0.05),levels of IL-1β,IL-6 and TNF-αalso increased(P<0.05),renal tubules and glomeruli in kidney tissue were both significantly damaged,accompanied by a large number of inflammatory cell infiltration,and pathological score increased(P<0.05),while proportion of ANXA1 positive staining area was decreased(P<0.05);neutrophils identified by Giemsa staining and trypan blue staining had complete morphology and high activity;compared with control group,apoptosis rate of neutrophils in AKI group was decreased(P<0.05),and the positive rate of TUNEL was decreased(P<0.05).Compared with AKI group,levels of Scr and BUN in serum of rats in AKI+Ac2-26 group were decreased(P<0.05),levels of IL-1β,IL-6 and TNF-αalso decreased(P<0.05),pathological manifestations of renal tubules and glomeruli in renal tissue were significantly reduced,and pathological score was reduced(P<0.05),while the proportion of ANXA1 positive staining area was increased(P<0.05),at the same time,apoptosis rate of rat neu-trophils was increased(P<0.05),positive rate of TUNEL was also increased(P<0.05).Conclusion:ANXA1 peptidomimetic Ac2-26 can increase expression of ANXA1 in kidney tissue of AKI in septic rats,promote neutrophil apoptosis,and have a protective effect on kidney tissue damage in rats caused by sepsis.
作者
黄承
蒲运刚
田仁富
杨先芹
张莉
HUANG Cheng;PU Yungang;TIAN Renfu;YANG Xianqin;ZHANG Li(Department of Critical Care Medicine,Enshi Central Hospital,Enshi 445000,China;Emergency ICU,Enshi Tujia and Miao Autonomous Prefecture Central Hospital,Enshi 445000,China;Department of Critical Care Medicine,Hubei Cancer Hospital,Wuhan 430000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2024年第6期1160-1165,共6页
Chinese Journal of Immunology
基金
湖北省卫生健康委2019年度第一批联合基金立项项目(WJ2019H130)。
