摘要
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45min后再灌注24h；硫氢化钠（NariS）预处理组：在缺血再灌注前通过左肾动脉持续注射NariS（300nmol／min）15min。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KB（NF—KB）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NODl、NOD2蛋白表达显著增加，细胞核内NF．KBP65表达及其靶基因MCP-1、IL-lβ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NODl、NOD2蛋白表达（P＜0．05），下调NF。KBP65的核转位（P＜0．05）以及局部MCP-1、IL．1B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。
Objective To investigate whether the nod-like receptor （NLR） pathway is involved in protection of hydrogen sulfide （H2S） preconditioning during renal isehemia reperfusion. Methods Male Wistar rats were randomly divided into 3 groups： sham operation （Sham） group, renal isehemia/ reperfusion （I/R） group subjected to occlusion of left renal pedicle for 45 min then reperfusion for 24 hours, and sodium hydrosulfide （NariS） preconditioning group with continuous infusion of Naris （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 - KB 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 stainingrevealed more apoptotic cells in risk zone with the activation of caspase-1 of I/R- treated kidney（P 〈 0.01）. NariS preconditioning reversed I/R-induced increase in the expression of NOD1 and NOD2（P〈0.05）. NariS preconditioning also reduced I/R- induced activation of NF- KB P65 （P〈0.05） and upregulation of MCP-1 and IL-113 （P〈0.01）. Moreover, NariS 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.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2014年第8期604-608,共5页
Chinese Journal of Nephrology
基金
山西省高等学校科技创新项目（20131106）
新疆第六师五家渠科技项目（1422）
