丹参酮Ⅱ_A治疗肾缺血-再灌注损伤及对缺氧诱导因子-1α表达的影响

Tanshinone II A treatment renal ischemia-reperfusion injury and effect the expression of hypoxia inducible factor-lalpha

目的探讨丹参酮ⅡA治疗肾缺血-再灌注损伤及其对缺氧诱导因子-1α(HIF-1α)表达的影响。方法选用Wistar大鼠66只,随机分为正常组、缺血组、缺血-再灌注组(1h、3h、5h)、丹参酮ⅡA组(1h、3h、5h)及假手术组(1h、3h、5h)。每组6只。各组均先切除右肾,取左肾研究。取左肾部分皮髓质用4%的多聚甲醛固定,进行切片HE染色及免疫组化检测各组肾HIF-1α蛋白的表达水平,另一部分采用荧光定量PCR检测各组肾HIF-1α mRNA的表达水平。取左肾后立即通过下腔静脉采血,检测MDA、TNF-α、Urea及Scr。结果 (1)Urea、HIF-1α蛋白及HIF-1α mRNA分别在假手术组1h、3h、5h增高,与正常组比较,差异有显著意义(P<0.05)。(2)Urea、Scr、MDA、TNF-α分别在正常组、缺血组、缺血-再灌注组(1h、3h、5h)增高(P<0.05)。HIF-1α mRNA在缺血组呈高表达,缺血-再灌注组(1h、3h、5h)呈低表达。HIF-1α蛋白在缺血组和缺血-再灌注组(1h、3h、5h)均呈高表达。(3)Urea、Scr、MDA、TNF-α、HIF-1α蛋白表达水平在丹参酮ⅡA组(1h、3h、5h)均较缺血-再灌注组降低(P<0.05),而HIF-1α mRNA表达水平在丹参酮ⅡA组(1h、3h、5h)增高(P<0.05)。(4)HE染色:与正常组比较,假手术组、缺血组变化不明显,其余各组均有不同程度肾小球、肾间质充血,肾小管上皮细胞水肿、细胞内玻璃样变及脂肪变性。缺血-再灌注组肾小管腔内蛋白管型明显。丹参酮ⅡA组仅个别标本存在蛋白管型,水肿减轻。结论丹参酮ⅡA可减轻肾缺血-再灌注损伤,可增强HIF-1α mRNA的表达和降低HIF-1α蛋白的表达。

Objective Approach the therapy of tanshinone ⅡA（Tan II a） to treat renal ischemia-reperfusion injury and effect the expression of hypoxia inducible factor-lalpha（hqF-ltz）. Methods 66 of Wistar rats were adopted and divided them into normal group,ischemia group,ischemia-reperfusion 1 h,3 h and 5 h group,Tan ⅡA Ih,3 h and 5 h group and sham operation 1 h,3 h and 5 h group,randomly. Every group was 6 and every rat was taken out the right renal before experiment and remaining the left one to research. A piece of cortico-medulla was cut out to make paraffin block by 4% Polyoxymethylene to fix in every left renal. The HE staining was carried out through section to the paraffin block and the expression of HIF-1α protein was detected through the paraffin section by immunohistochemistry. The other moiety was stored by subambient temperature to extract the total RNA and follow to detect the expression of HIF-1α mRNA through real-time PCR. After resection the left renal get blood through pricking inferior vena cave to determine MDA,TNF-α,Urea and Scr. Results （1）it was statistical significance（P0.05） compare Urea,the expression of HIF-1α protein and HIF-1α mRNA in the sham operation group 5 h,3 h and Ih with the normal group.（2）Urea,Scr,MDA,TNF-α were increase in the normal group,ischemia group and ischemia-reperfusion 1 h,3 h,5 h group in turn（P0.05）. It was high-expression about HIF-1α mRNA in ischemia group and the low-expression in ischemia-reperfusion 1 h,3 h,5 h group.High-expression about HIF-1α protein was found in ischemia group and ischemia-reperfusion 1 h,3 h,5 h group.（3）ll of Urea,Scr,MDA,TNF-α,the expression of HIF-1α protein were lower in Tan ⅡA 1 h,3 h,5 h group,however the expression of HIF-1 ct mRNA were higher in Tm ⅡA 1 h,3 h,5 h group than ischemia-re -perfusion every group（P0.05）. FHE staining：it was no much change in the sham and ischemia group compare with normal group.The other groups were found different extent hyperemia in glomcrulus and renal interstitium,and different extent cellularedema in renal tubular epithelial cell. It was obviously that Protein cast in renal tubules of the ischemia-reperfusion every group. But it was individual in Tan II,every group and the ceUularedema abatement. Conclusion Tan ⅡA treatment could attenuate renal IRI and could increase expression of HIF-lct mRNA,lower the expression of HIF-1 tx protein.