冷缺血损伤对大鼠供肾转化生长因子β1表达的影响

Effects of cold ischemia injury on expression of transforming growth factor-β1 in rat kidney

目的探讨冷保存对转化生长因子β1(TGFβ1)在大鼠肾小管上皮细胞表达的影响。方法选择Wistar大鼠40只,采取原位灌注整块肾脏切取,HC-A(肾脏保护液)灌注,在0～4℃下分别保存0,12,24,36和48h,采用免疫组织化学术,原位杂交术分别检测肾小管上皮细胞TGFβ1表达水平。结果在0h组,12h组大鼠肾小管上皮细胞中TGFβ1极少表达,两组TGFβ1蛋白,mRNA平均阳性单位(PU值)分别为(6.37±2.77),(5.29±2.15);(6.11±1.82),(5.90±2.40);差异均无显著性(P>0.05)。随着冷缺血时间延长至24、36及48h,TGFβ1蛋白,mRNA表达逐渐增加[24h组PU值分别为(10.20±3.27),(11.32±3.34);36h组PU值分别为(13.84±3.91),(15.47±4.21);48h组PU值分别为(17.17±3.96),(19.01±3.53)],各组间比较差异有显著性(P<0.05),各组与0h,12h组相比较差异亦有显著性(P<0.05)。结论冷缺血损伤促进大鼠肾脏转化生长因子-β1表达增加,促进细胞外基质的合成。

[Objective] To investigate the effect of cryopreservation on the expression of transforming growth factor β1（TGFβ1） in the renal tubular epithelial. [Methods] A total of 40 Wistar rats were used in this study. In terms of the time of cryopreservation of donor kidney, the 40 Wistar rats were divided into 5 groups： 0 h group （control group）, 12 h group, 24 h group, 36 h group, 48 h group. A block removal of donor kidney with in situ perfusion of cooling HC-A preservation solution was adopted. The rat kidney was preserved 0, 12, 24, 36 and 48 h at 0~4℃, respectively. The expression of TGF-β1 protein and mRNA in the proximal tubular epithelial cells were detected by in situ hybridization and immunohistochemistry analysis. Value of positive unit was calculated by image analysator. [Results] Only a small amount of TGF-β1 protein and mRNA were expressed in CP 0 h group and CP 12 h group, TGF-β1 protein PU value was （6.37±2.77） and （6.11±1.82） （P 〉0.05）, respectively. TGF-β1 mRNA PU value was （5.29±2.15） and （5.90±2.40） （P 〉0.05）, respectively. As the cold ischemia time prolonged, the protein PU value of TGF-β1 is increased at CP 24 h group （10.20±3.27）, CP 36 h group （13.84±3.91）, CP 48 h group （17.17±3.96）, respectively. There is significant difference between these groups and when compared with CP 0 h and 12 h group （P 〈0.05）. The mRNA PU value of TGF-β1 was also increased at CP 24 h group （11.32±3.34）, CP 36 h group （15.47±4.21）, CP 48 h group （19.01±3.53）, respectively. There is significant difference between these groups and when compared with CP 0 h and 12 h group （P 〈0.05）. [Conclusion] TGFβ1 expression is increased with severe cold ischemia injury, which accelerates the formation of extracellular matrix.