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目的探讨中华眼镜蛇毒(CCV)对大鼠肾缺血再灌注(I/R)损伤的肾保护作用及其机制。方法雄性SD大鼠32只,随机分为4组。Ⅰ、Ⅱ组建立肾I/R模型,分别于再灌注前0.5h、12h腹腔注射0.1%CCV(0.4mg/kg);Ⅲ组制备肾I/R模型作病理对照:Ⅳ组为假手术组。应用苦味酸法和二乙酰一肟法分别测定大鼠缺血前、再灌注24h的Scr和BUN值。用免疫比浊法测定再灌注0、0.5、2、24h的血清补体C3水平。HE染色光镜下观察肾组织损伤形态学改变,并利用原位凋亡检测法(TUNEL)检测细胞凋亡情况。结果再灌注后24h的Scr和BUN值在Ⅱ组明显降低,与Ⅰ、Ⅲ组相比差异有统计学意义(P<0.05)。补体C3水平在Ⅱ组于再灌注0h显著降低,与其他组0h时比较,差异均有统计学意义(P<0.05):Ⅰ、Ⅲ组C3于再灌注2h明显下降,与再灌注0h比较,差异有统计学意义(P<0.05)。HE染色可见Ⅰ、Ⅲ组损伤较重,以近端小管变性,坏死为主;Ⅱ组病变明显减轻。在Ⅲ和Ⅰ组均可见大量TUNEL阳性细胞,而在Ⅱ组其数量与上两组相比显著减少(P<0.05)。结论肾I/R可明显引起肾组织损伤和功能损害。再灌注前12h用CCV预处理大鼠,能够明显降低补体C3,从而有效抑制补体介导的肾I/R损伤。
Objective To investigate the protective effect of Chinese cobra venom (CCV) on renal ischemia-reperfusion (I/R) injury in rats and its mechanism. Methods Thirty-two male SD rats were randomly divided into 4 groups. Kidney I/R models were established in groups I and II, and 0.1% CCV (0.4 mg/kg) was injected intraperitoneally 0.5h and 12h before reperfusion respectively; III group was used to establish renal I/R model for pathological control: IV group was sham-operated group . The levels of Scr and BUN in rats before and after reperfusion were determined by picric acid method and diacetyl-leptin method. Serum complement C3 levels at 0, 0.5, 2, 24 h after reperfusion were measured by immunonephelometry. HE staining was performed under light microscope to observe the morphological changes of renal tissue injury, and TUNEL was used to detect cell apoptosis. Results The Scr and BUN values at 24 h after reperfusion were significantly decreased in group II, and there were significant differences compared with group I and III (P<0.05). The level of complement C3 was significantly lower in group II at 0 h after reperfusion, compared with 0 h in other groups, the difference was statistically significant (P<0.05): C3 in group I and III was significantly decreased at 2 h after reperfusion, compared with 0 h after reperfusion. The difference was statistically significant (P<0.05). Hematoxylin-eosin staining showed that group I and group III had more severe lesions, with degeneration and necrosis of proximal tubules. The lesions of group II were significantly reduced. A large number of TUNEL-positive cells were seen in group III and I, while the number of TUNEL-positive cells in group II was significantly reduced compared with the two groups (P<0.05). Conclusion Renal I/R can obviously cause renal tissue damage and functional impairment. Pretreatment with CCV 12 h before reperfusion could significantly reduce complement C3 and effectively inhibit complement-mediated renal I/R injury.