外源性硫化氢在大鼠肾脏缺血再灌注损伤中的保护作用

Protective effect of exogenous hydrogen sulfide on renal ischemia reperfusion injury in rats

目的观察不同剂量外源性硫化氢（H2S）供体硫氢化钠对大鼠肾脏缺血再灌注损伤（IRI）的保护作用。方法健康雄性Wistar大鼠28只随机分为4组，即假手术组（Sham）、肾缺血再灌注（IR）组、硫氢化钠（NaHS）高剂量组、硫氢化钠低剂量组。大鼠右肾切除后，以NaHS作为硫化氢的供体，NaHS高、低剂量组分别经左肾动脉插管，按照1．5μmol／min、300nmol／min的剂量连续15min给药，假手术组及IR组给予同体积生理盐水。停药5min后，NaHS组和IR组用无损伤微动脉夹夹闭左侧肾蒂45min后解除阻断，建立大鼠急性IRI模型，假手术组不夹闭左肾动脉，其他操作同模型组。于肾脏恢复血流24h时留取血和肾组织标本，检测血清尿素氮（BUN）、血肌酐（Scr）；半定量分析肾脏病理损伤；检测肾组织H2S生成率；采用实时定量PCR法检测胱硫醚-8-合成酶（CBS）、胱硫醚-1-裂解酶（CSE）mRNA表达。结果与假手术组相比，IR组H2s生成率显著降低（P〈0．01）；CBS、CSEmRNA表达显著下降（P〈0．01）；Scr、BUN显著升高（P〈0．01）；肾脏病理表现为急性肾小管坏死，且最严重。与IR组相比，NaHS预处理组H2s生成率升高（P〈0．05）；CBS、CSEmRNA表达升高（P〈0．01）；Scr、BUN降低（P〈0．01）；病理损伤明显减轻。NaHS两个剂量组之间差异无统计学意义。结论外源性H2S对大鼠IRI具有保护作用。

Objective To observe the protective effect of different doses sodium hydrosulfide （NariS） as a donor of hydrogen sulfide on renal ischemia reperfusion injury （IRI）. Methods Twenty-eight healthy male Wistar rats were randomly divided into 4 groups, namely sham operation （Sham） group （n =7）, renal ischemia reperfusion （IR） group （n =7）, sodium hydrosulfide （NariS） high dose group （n=7）, sodium hydrosulfide low dose group （n=7）. After excision of the fight kidney, two dose Naris group （300 nmol/min, 1.5 μmol/min） received 15- minute continuous administration via left renal artery. Sham group and IR group received same volume of saline. Five minutes after stopping drug, the left renal pedicle in Naris group and IR group was clipped with no damage arteriole occlusion. After 45 minutes blockade, a model of acute renal ischemia reperfusion injury was established. Sham group underwent the same procedure as model groups without clamping the left renal artery. Specimens of renal tissue and blood were harvested at 24-hour after blood flow restore in the kidney. BUN and Set were measured. Kidney pathological damage was semi-quantitatively analyzed. Production of H2S in renal tissue was detected. Expression of cystathionine T lyase （CSE） and cystathionine 13 synthase （CBS） mRNA in kidney tissue was examined by real-time PCR. Results Compared with sham group, BUN and Scr increased significantly （P〈O.01）, kidney tissue H2S production decreased significantly （P〈0.01）, mRNA expression of CSE and CBS down-regulated significantly （P〈0.O1） in IR group. The kidney pathology of sham group was normal, while acute tubular necrosis was found in IR group. Compared with IR group, BUN and Scr decreased significantly （P〈O.01）, kidney tissue H2S production increased significantly （P〈0.05）, mRNA expression of CSE and CBS up-regulated significantly （P〈 0.01） in NariS groups. Pathological damage of acute tubular necrosis was significantly improved in NariS pretreatment group. There was no significant difference between two NariS doses groups. Conclusion Hydrogen sulfide has a protective effect on renal IRI.