中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2014年
8期
604-608
,共5页
覃志成%石媛媛%闫燕%吴莺莺%刘文丽%李光远%李荣山
覃誌成%石媛媛%閆燕%吳鶯鶯%劉文麗%李光遠%李榮山
담지성%석원원%염연%오앵앵%류문려%리광원%리영산
硫化氢%再灌注损伤,肾脏%NOD样受体
硫化氫%再灌註損傷,腎髒%NOD樣受體
류화경%재관주손상,신장%NOD양수체
Hydrogen sulfide%Ischemia-reperfusion,Kidney%NLR
目的 观察NOD样受体途径是否参与硫化氢(H2S)预处理对大鼠肾脏缺血再灌注损伤(IRI)的保护作用.方法 雄性Wistar大鼠被随机分为3组:假手术(Sham)组;肾脏缺血再灌注(I/R)组:夹闭左侧肾蒂45 min后再灌注24 h;硫氢化钠(NaHS)预处理组:在缺血再灌注前通过左肾动脉持续注射NaHS(300 nmol/min) 15 min.HE染色观察肾组织学改变;Western印迹法检测肾组织核苷酸结合寡聚化结构域(NOD)1、NOD2、半胱氨酸蛋白酶(Caspase)1及细胞核因子κB(NF-κB)蛋白的表达;免疫组织化学法检测单核细胞趋化蛋白1(MCP-1)、白细胞介素(IL)-1β的表达变化;TUNEL法检测肾组织细胞凋亡.结果 与Sham组比较,I/R组大鼠肾组织NOD1、NOD2蛋白表达显著增加,细胞核内NF-κB P65表达及其靶基因MCP-1、IL-1β的蛋白表达显著升高(均P＜ 0.01).Caspase-1蛋白表达增加,HE染色发现急性肾小管坏死,TUNEL染色显示缺血危险区凋亡细胞数目显著增加(均P＜0.01).NaHS预处理可逆转I/R诱导的肾组织损伤以及NOD1、NOD2蛋白表达(P＜0.05),下调NF-κB P65的核转位(P＜0.05)以及局部MCP-1、IL-1β表达(P＜0.01).同时,NaHS预处理可减少Caspase-1的活化和细胞凋亡数.结论 硫化氢预处理通过NOD样受体依赖的炎症途径减轻肾缺血再灌注损伤.
目的 觀察NOD樣受體途徑是否參與硫化氫(H2S)預處理對大鼠腎髒缺血再灌註損傷(IRI)的保護作用.方法 雄性Wistar大鼠被隨機分為3組:假手術(Sham)組;腎髒缺血再灌註(I/R)組:夾閉左側腎蒂45 min後再灌註24 h;硫氫化鈉(NaHS)預處理組:在缺血再灌註前通過左腎動脈持續註射NaHS(300 nmol/min) 15 min.HE染色觀察腎組織學改變;Western印跡法檢測腎組織覈苷痠結閤寡聚化結構域(NOD)1、NOD2、半胱氨痠蛋白酶(Caspase)1及細胞覈因子κB(NF-κB)蛋白的錶達;免疫組織化學法檢測單覈細胞趨化蛋白1(MCP-1)、白細胞介素(IL)-1β的錶達變化;TUNEL法檢測腎組織細胞凋亡.結果 與Sham組比較,I/R組大鼠腎組織NOD1、NOD2蛋白錶達顯著增加,細胞覈內NF-κB P65錶達及其靶基因MCP-1、IL-1β的蛋白錶達顯著升高(均P＜ 0.01).Caspase-1蛋白錶達增加,HE染色髮現急性腎小管壞死,TUNEL染色顯示缺血危險區凋亡細胞數目顯著增加(均P＜0.01).NaHS預處理可逆轉I/R誘導的腎組織損傷以及NOD1、NOD2蛋白錶達(P＜0.05),下調NF-κB P65的覈轉位(P＜0.05)以及跼部MCP-1、IL-1β錶達(P＜0.01).同時,NaHS預處理可減少Caspase-1的活化和細胞凋亡數.結論 硫化氫預處理通過NOD樣受體依賴的炎癥途徑減輕腎缺血再灌註損傷.
목적 관찰NOD양수체도경시부삼여류화경(H2S)예처리대대서신장결혈재관주손상(IRI)적보호작용.방법 웅성Wistar대서피수궤분위3조:가수술(Sham)조;신장결혈재관주(I/R)조:협폐좌측신체45 min후재관주24 h;류경화납(NaHS)예처리조:재결혈재관주전통과좌신동맥지속주사NaHS(300 nmol/min) 15 min.HE염색관찰신조직학개변;Western인적법검측신조직핵감산결합과취화결구역(NOD)1、NOD2、반광안산단백매(Caspase)1급세포핵인자κB(NF-κB)단백적표체;면역조직화학법검측단핵세포추화단백1(MCP-1)、백세포개소(IL)-1β적표체변화;TUNEL법검측신조직세포조망.결과 여Sham조비교,I/R조대서신조직NOD1、NOD2단백표체현저증가,세포핵내NF-κB P65표체급기파기인MCP-1、IL-1β적단백표체현저승고(균P＜ 0.01).Caspase-1단백표체증가,HE염색발현급성신소관배사,TUNEL염색현시결혈위험구조망세포수목현저증가(균P＜0.01).NaHS예처리가역전I/R유도적신조직손상이급NOD1、NOD2단백표체(P＜0.05),하조NF-κB P65적핵전위(P＜0.05)이급국부MCP-1、IL-1β표체(P＜0.01).동시,NaHS예처리가감소Caspase-1적활화화세포조망수.결론 류화경예처리통과NOD양수체의뢰적염증도경감경신결혈재관주손상.
Objective To investigate whether the nod-like receptor (NLR) pathway is involved in protection of hydrogen sulfide (H2S) preconditioning during renal ischemia reperfusion.Methods Male Wistar rats were randomly divided into 3 groups:sham operation (Sham) group,renal ischemia/ reperfusion (I/R) group subjected to occlusion of left renal pedicle for 45 min then reperfusion for 24 hours,and sodium hydrosulfide (NaHS) preconditioning group with continuous infusion of NaHS (300 nmol/min) by left renal artery for 15 min before I/R treatment.Renal injuries were evaluated by HE staining.The protein levels of NOD1,NOD2,nuclear NF-κB P65 and caspase-1 were analyzed by Western blot assay.The protein level of MCP-1 and IL-1β expressions was determined by immunohistochemical staining assay.Cell apoptosis were evaluated by Tunel staining assay.Results In I/R group,the renal NOD1 and NOD2 protein expressions were upregulated.Moreover,the nuclear NF-κB P65 expression was also elevated with an increase in its target genes-MCP-1 and IL-1β (All P ＜ 0.01).HE staining revealed the existence of acute tubular necrosis in I/R kidney.TUNEL staining revealed more apoptotic cells in risk zone with the activation of caspase-1 of I/R-treated kidney(P ＜0.01).NaHS preconditioning reversed I/R-induced increase in the expression of NOD1 and NOD2(P ＜0.05).NaHS preconditioning also reduced I/R-induced activation of NF-κB P65 (P ＜ 0.05) and upregulation of MCP-1 and IL-1β (P ＜ 0.01).Moreover,NaHS preconditioning attenuated inflammation,repressed caspase-1 activation and reduced apoptotic cells after I/R.Conclusion Hydrogen sulfide preconditioning can alleviate renal ischemia/reperfusion injury by Nod-like receptor dependent on inflammatory pathway.
