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目的探讨大鼠供肾在不同冷保存时间,其肾小管近端上皮细胞结缔组织生长因子(connective tissue growth factor,CTGF)的表达。方法健康SPF级Wistar雄性大鼠40只,体重230～250g。采取原位灌注切取整块双侧肾脏,置于0～4℃高渗枸橼酸盐嘌呤肾保存液中保存。根据保存时间不同,分为0、12、24、36、48h组(n=8)。采用免疫组织化学、原位杂交方法分别检测肾小管近端上皮细胞CTGF表达水平。结果在0、12h组大鼠供肾肾小管近端上皮细胞中CTGF极少表达,经图像分析两组CTGF蛋白阳性单位值(positive unit,PU)分别为5.91±2.30、5.57±2.40;CTGF mRNA PU值分别6.24±2.79、6.51±2.43,差异均无统计学意义(P>0.05)。随着冷缺血时间延长,24、36、48h组CTGF蛋白和CTGF mRNA表达均逐渐增加,24h组PU值分别为10.25±2.92、15.24±3.95;36h组分别为14.31±2.83、19.20±4.73;48h组分别为18.11±3.94、23.09±4.40;各组间比较差异均有统计学意义(P<0.05),与0、12h组比较差异亦均有统计学意义(P<0.05)。结论冷缺血损伤促进大鼠肾脏CTGF表达增加,可能在肾小管上皮细胞损伤再生与修复中起重要作用。
Objective To investigate the expression of connective tissue growth factor (CTGF) in proximal tubular epithelial cells of rats during different cold storage periods. Methods Forty SPF Wistar male rats weighing 230-250 g were used. In situ perfusion cut the entire bilateral kidneys, placed 0 ~ 4 ℃ hypertonic citrate purine kidney preservation solution stored. According to the different storage time, divided into 0,12,24,36,48 h group (n = 8). Immunohistochemistry and in situ hybridization were used to detect the expression of CTGF in proximal tubular epithelial cells. Results The expression of CTGF protein in proximal renal proximal tubular epithelial cells of 0, 12h rats was poorly expressed. CTGF protein positive units (PUs) of the two groups were 5.91 ± 2.30 and 5.57 ± 2.40, respectively. CTGF mRNA PU value were 6.24 ± 2.79,6.51 ± 2.43, the difference was not statistically significant (P> 0.05). With the prolongation of cold ischemia, the expression of CTGF protein and CTGF mRNA in 24, 36 and 48h groups increased gradually, the PU values in 24h group were 10.25 ± 2.92 and 15.24 ± 3.95, respectively, and those in 36h group were 14.31 ± 2.83 and 19.20 ± 4.73 respectively; 48h group were 18.11 ± 3.94 and 23.09 ± 4.40, respectively. There was significant difference between the two groups (P <0.05), but there were also significant differences between the two groups (P <0.05). Conclusion Cold ischemia injury promotes the expression of CTGF in rat kidney, which may play an important role in the regeneration and repair of renal tubular epithelial cells.