外源性硫化氢对大鼠梗阻肾组织中血管紧张素Ⅱ和PCNA及TGF-β_1表达的影响

Effects of exogenous hydrogen sulfide on the expressions of angiotensin Ⅱ and proliferating cell nuclear antigen of tubulointerstitial fibrosis in rats with unilateral ureteral obstruction

目的肾小管间质纤维化(TIF)是反映肾功能下降严重程度和及判断预后的最重要指标,硫化氢(H_2S)在维持肾的正常功能和多种肾损伤疾病中都起着关键作用。文中通过建立单侧输尿管梗阻模型(UUO)观察外源性补充硫化氢(H_2S)供体硫氢化钠(Na HS)对单侧输尿管梗阻(UUO)大鼠肾组织中血管紧张素Ⅱ(AngⅡ)、增殖细胞核抗原(PCNA)和转化生长因子-β1(TGF-β1)表达的影响。方法单侧输尿管梗阻建立TIF动物模型,96只SD大鼠随机数字表法分为假手术组、模型组,以及Na HS低、高剂量组,每组24只。模型组大鼠行左侧输尿管结扎,假手术组大鼠仅游离输尿管而不进行结扎。Na HS低、高剂量组治疗组大鼠在UUO制备后分别给予腹腔注射Na HS 1.4μmol/kg(0.2μmol/m L)和7μmol/kg(1μmol/m L),假手术组及模型组分别腹腔注射等量等渗盐水。术后第7、14、21天分别进行HE、Masson染色和免疫组化染色。测定大鼠肾间质纤维化相对面积以及肾组织AngⅡ、TGF-β1、PCNA相对表达。结果假手术组各时间点AngⅡ、PCNA和TGF-β1在肾小管间质微量表达。模型组较假手术组7、14、21 d AngⅡ表达升高(6.27±0.78 vs 1.09±0.75,8.94±0.31 vs 1.20±0.06,13.75±0.39 vs1.34±0.69;P<0.01)、PCNA表达升高(82.81±5.36 vs 4.63±0.83,101.00±4.3 vs 8.57±1.09,118.11±4.70 vs 13.15±1.43;P<0.01),TGF-β1表达亦升高(9.18±0.83 vs 0.98±0.14,16.87±0.77 vs 1.85±0.20,20.29±0.89 vs 2.43±0.14,P<0.01)。与模型组比较,Na HS低、高剂量组AngⅡ、PCNA和TGF-β1表达均降低(P<0.05)。但Na HS低、高剂量组之间差异无统计学意义(P>0.05)。结论外源性补充H_2S可部分通过下调AngⅡ、PCNA和TGF-β1的表达延缓TIF的进展。

Objective Tubulointerstitial fibrosis （TIF） is the most important marker reflecting the degree of renal function decline and prognosis and hydrogen sulfide （H2 S） is crucial in maintaining normal renal function and many diseases of renal injury. The aim of the article was to investigate the effects of exogenous H2S on the expressions of angiotensin Ⅱ （Ang Ⅱ ）, proliferating cell nuclear antigen （PCNA） and transforming growth factor beta-1 （TGF-β1） in rats with unilateral ureteral obstruction （UUO）. Methods TIF rat model was built with UUO. Ninety-six SD rats were randomly divided into four groups： sham operation group, model group, UUO＋low-dose NariS treatment group （low dose group） and UUO＋high-dose Naris treatment group （high dose group） （n= 24, respectively）. Rats in model group were treated with left-side ureteral obstruction and ureteral separation without obstruction was done in sham operation group. UUO rats in two treatment groups were injected intraperitoneally with two different doses of sodium hydrosulfide （ NariS, donor of endogenous H2 S）, respectively. HE and Masson staining and immunohistochemical staining were performed at the 7 d, 14 d and 21 d, respectively. Results In sham operation group, the expressions of Ang Ⅱ, PCNA, and TGF-β1were found in microamount in tubulointerstitium at each time points. Compared with sham operation group, the expressions of Ang Ⅱ , PCNA and TGF-β1 in model group increased（P〈0.01）. While in comparison to model group, the expressions of Ang Ⅱ , PCNA and TGF-β1 decreased in low dose group and high dose group, but no significant difference was found between low dose group and high dose group. Conclusion Exogenous H2S supplementation can attenuate TIF partly via downregulating the expressions of Ang Ⅱ, PCNA and TGF-β1.