丹参酮ⅡA保护大鼠肾移植术后缺血再灌注损伤的机制研究

Tanshinone ⅡA protection mechanism of renal transplantation in rats after ischemia reperfusion injury

目的观察丹参酮ⅡA对大鼠同种异体肾移植术后肾脏保护性因子Klotho基因和氧化应激对肾功能等的影响,分析其对肾缺血/再灌注损伤的保护作用机制。方法 SD大鼠48只,分为假手术组、缺血/再灌注组、丹参酮A组和B组,采用同种异体左肾移植手术。术后丹参酮A、B组分别用5%和10%丹参酮按5 m L/kg灌胃治疗7 d,假手术组和缺血/再灌注组按5 m L/kg体积分别灌服生理盐水。采用RT-PCR法检测肾组织Klotho基因表达,从尾静脉采血测定4组大鼠肾功能:肌酐(Cr)、尿素氮(BUN)、胱抑素C(Cys C)]指标。检测肾组织匀浆中氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)和血清丙二醛(MDA)。结果丹参酮A组和B组大鼠肾移植术后BUN、Cr、Cys C显著降低,肾组织匀浆中MDA降低、SOD提高,Klotho蛋白和lotho-m RNA明显上调,与缺血/再灌注组比较,P<0.05。丹参酮A组和B组组间比较,P>0.05。结论丹参酮ⅡA在肾移植术后可提高SOD活性、抑制MDA的产生,上调Klotho蛋白和lotho-m RNA表达,抑制肾小管上皮细胞凋亡而达到肾缺血/再灌注损伤的保护作用。

Objective To observe the effect of tanshinone ⅡA on rats＇ kidney after taking allograft transplantation and renal, protective factor Klotho gene and oxidative stress on renal function and analysis of the renal ischemia/reperfusion injury on protection mechanism. Methods 48 SD rats were randomly divided into sham operation group, ischemia/reperfusion group and tanshinone A group and B group, the whole of which took the left renal allograft transplantation. After operations tanshinone A group and B group were respectively treated to lavage for 5% and 10% by Tanshinone 5m L/kg for 7 days（d）. According to the volume fraction, sham operation group and ischemia/reperfusion group were treated of 5m L/kg normal saline. The expression of Klotho gene in renal tissue was detected by RT-PCR method.Collected vein blood from the tail of four rats in order have determination of renal function [creatinine（Cr）; urea nitrogen（BUN） and cystatin C（Cys C）three. Then had detection of superoxide dismutase in kidney homogenates of enzyme（SOD）, lactate dehydrogenase（LDH） and serum malondialdehyde（MDA）. Results After renal transplantation,rats of Tanshinone A group and B group, BUN, Cr and Cys C decreased significantly, the kidney tissue in MDA decreased, SOD increased, Klotho protein and lotho-m RNA was up-regulated, compared with ischemia/reperfusion group, P 0.05. When having comparison between tanshinone A group and B group, P 0.05. Conclusion Tanshinone ⅡA can improve SOD＇s activity and inhibit the production of MDA after renal transplantation, so as the expression of Klotho and lotho-m RNA, thus inhibit the apoptosis of renal tubular epithelial cells to protect renal ischemia/reperfusion injury.