内源性硫化氢通过抑制NOD1信号通路在缺血性急性肾损伤中保护作用的研究  

Protective role of endogenous hydrogen sulfide in ischemic AKI by inhibiting NOD1 signal pathway

作　　者：王佳丽[1] 李光远 穆丽娜 张哲 刘文丽 王军霞 马芳 覃志成[1] 

机构地区：[1]山西医科大学第二医院肾内科,山西030001 [2]新疆五家渠人民医院肾内科,新疆831399

出　　处：《中华肾病研究电子杂志》2017年第2期73-78,共6页Chinese Journal of Kidney Disease Investigation（Electronic Edition）

基　　金：新疆兵团应用基础研究项目(2015AG017)

摘　　要：目的观察在缺血性急性肾损伤中抑制内源性硫化氢产生是否可通过激活NOD1信号通路加重肾间质炎症反应与细胞凋亡。方法将雄性Wistar大鼠随机分为4组:假手术(Sham)组、肾脏缺血再灌注(I/R)组、I/R+炔丙基甘氨酸(PAG)组、I/R+羟氨(HA)组。通过Western印记法分别对肾组织模式识别受体NOD1、半胱氨酸蛋白酶(caspase-1)及细胞核因子-κB(NF-κB)蛋白的表达进行检测;实时定量PCR法对NOD1的mRNA表达进行检测;HE染色法观察肾脏组织学改变;免疫组织化学法检测肾组织TNF-ɑ的表达;TUNEL法检测肾组织细胞凋亡。采用SPSS 22.0统计软包对实验数据进行统计学处理,P<0.05被认为差异有统计学意义。结果与Sham组比较,I/R组大鼠肾组织NOD1、caspase-1、NF-κB、TNF-ɑ蛋白表达增加(t=17.81,t=7.78,t=12.08,t=10.3,P﹤0.05),NOD1mRNA表达增加(t=8.73,P﹤0.05),HE染色后者表现为急性肾小管坏死,肾小管损伤评分明显增加(t=11.0,P﹤0.05),TUNEL染色显示缺血区凋亡细胞数目增加(t=18.47,P﹤0.05)。与I/R组比较,I/R+PAG组NOD1、caspase-1、NF-κB、TNF-ɑ蛋白表达增加(t=12.51,t=8.81,t=5.88,t=11.04,P﹤0.05),NOD1 mRNA表达增加(t=8.11,P﹤0.05),HE染色后者表现为急性肾小管坏死,肾小管损伤评分明显增加(t=13,P﹤0.05),TUNEL染色显示缺血区凋亡细胞数目增加(t=16.41,P﹤0.05)。与I/R组比较,I/R+HA组的NOD1、caspase-1、NF-κB、TNF-ɑ蛋白表达增加(t=13.35,t=9.4,t=5.27,t=4.98,P﹤0.05);NOD1mRNA表达增加(t=8.94,P﹤0.05);HE染色后者表现为急性肾小管坏死,肾小管损伤评分明显增加(t=13,P﹤0.05);TUNEL染色显示缺血区凋亡细胞数目增加(t=10.77,P﹤0.05)。结论抑制硫化氢可激活NOD1样受体依赖的炎症途径,从而加重肾缺血再灌注损伤,即硫化氢对缺血性急性肾损伤有保护作用。Objective To observe whether restraining production of endogenous hydrogen sulfide in ischemic acute kidney injury can aggravate renal interstitial inflammation and cell apoptosis by activating NOD1 signal pathway. Methods Male Wistar rats were randomly divided into 4 groups： sham group,kidney ischemia/reperfusion（ I/R） group,kidney ischemia/reperfusion（ I/R） group ＋ propargyl glycine（ PAG） group,and kidney ischemia/reperfusion（ I/R） group ＋ hydroxyammonia（ HA） group. Western blotting detected the kidney tissue NOD1,caspase-1,and NF-κB proteins; Real-time quantitative PCR detected NOD1 mRNA; Immunohistochemistry detected the kidney TNF-ɑ expression; HE staining observed pathological change in kidneys; and TUNEL detected kidney tissue cells apoptosis. Results Compared with the sham group,the I/R group kidneys showed significantly increased proteins expression of NOD1（ t =17. 81,P = 0. 006）,caspase-1 protein expressions（ t = 7. 78,P = 0. 04）,NF-κB（ t = 12. 08,P = 0. 04）,and TNF-ɑ（ t = 10. 3, P = 0. 026）; NOD1 mRNA expression also increased significantly（ t =8. 73,P = 0. 007）; HE staining showed that the acute tubular necrosis,and renal tubular injury scores increased significantly（ t = 11. 0,P = 0. 04）; TUNEL dyeing showed that apoptotic cells increased significantly in ischemia zone（ t = 18. 47,P = 0. 023）. Compared with the I/R group,I/R ＋ PAG group showed significantly increased proteins expression of NOD1（ t = 12. 51,P = 0. 031）,caspase-1（ t = 8. 81,P = 0. 024）,NF-κB（ t = 5. 88,P = 0. 02）,and TNF-ɑ（ t = 11. 04,P = 0. 04）; NOD1 mRNA expression increased significantly（ t = 8. 11,P = 0. 001）; HE staining showed that the acute tubular necrosis and renal tubular injury scores increased significantly（ t = 13,P = 0. 03）; TUNEL dyeing showed that apoptotic cells increased significantly in ischemia zone（ t = 16. 41,P = 0. 04）. I/R ＋ HA group also showed significantly increased proteins expression of NOD1（ t = 13. 35,P = 0. 006�

关 键 词：硫化氢 缺血再灌注 NOD1受体 

分 类 号：R692[医药卫生—泌尿科学]