肾缺血/再灌注时肾组织损伤及细胞凋亡的实验研究

目的：检测肾缺血／再灌注不同时问点肾组织、功能损伤及细胞凋亡的变化，探讨细胞凋亡在肾缺血／再灌注损伤中发生的机制。方法：应用苦味酸法和二乙酰一肟反应法测定大鼠肾缺血／再灌注不同时间点血肌酐和尿素氮值检测肾功能变化，用HE染色光镜下观察缺血／再灌注石蜡包埋切片肾组织损伤形态学改变，用酚／氯仿抽提小片断DNA，在琼脂糖电泳上测定不同时间点DNA Ladder及利用原位凋亡检到法观察各时间点TUNEL阳性细胞数检测细胞凋亡情况。结果：肾功能检测发现，缺血45min后血肌酐和尿素氮值明显增高和正方对照差异显(P＜0.05)，再灌注早期上升缓慢，3h后再次增高，组内对比差异显(P＜0．05)。HE染色光镜下观察肾组织细胞以近端肾小管变性为主、坏死轻微，缺血／再灌注各时间点组织损伤变化与肾功能变化规律相一致。琼脂糖电泳和TUNEL检测在缺血45min再灌注3h时可测到DNA Ladder和TUNEL阳性细胞增加，再灌注12h细胞凋亡最明显。结论：肾缺血／再灌注可引起肾组织轻、中度的损伤和明显的肾功能损害；肾缺血／再灌注后期肾组织及功能损伤加重可能与细胞凋亡的发生有关。

The Studies of Renal Tissue Injure and Cell Apoptosis in Ischemic/Reperfused Kidney

Objective:To examine the renal histological and functional damage and mechanism of apoptosis in ischemic/reperfused kidney.Methods:Renal function was measured by Bun and Scr. The kidney samples were examined by HE stain under light microscopy. Apoptosis was detected by agarose gel electrophoresis and Terminal Deoxynucleotidyl Transferase-Mediated dUTP-Biotin In Situ Nick-End Labeling(TUNEL).Results:Significant histological damage and impaired renal function were found after 45 minutes of ischemia and 3 hours of reperfusion. Proximal and inner medulla tubular cells showed significant degeneration with minimal necrosis. Significant apoptosis of tubular cells was also noted, which is most obvious at 12 hour.Conclusion:Renal histological and functional injury following ischemia/reperfusion may be associated with apoptosis of cells.