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目的:观察缺血再灌注损伤(ischemia/reperfusioninjury,IRI)后,低龄和高龄p53(+/+)和p53(-/-)鼠肾小管上皮细胞的演变,探讨p53基因对IRI后肾小管上皮细胞演变的影响。方法:分别建立低龄(2月龄)和高龄(12月龄)p53(+/+)和p53(-/-)鼠左肾IRI模型。于IRI后0d、1d、3d、7d、1月、3月、6月,用HE染色观察肾小管组织学变化,免疫组织化学法检测肾小管上皮细胞增殖细胞核抗原(prolifera-tingcellnuclearantigen,PCNA)的表达,组织化学染色观察肾小管上皮细胞衰老相关β-半乳糖苷酶(SA-β-gal)的活性,用脱氧核苷酸末端转移酶介导的缺口末端标记(terminaldeoxynu-cleotidyltransferase-mediateddUTP-biotininsitunick-endlabe-ling,TUNEL)法检测肾小管上皮细胞凋亡。结果:IRI后0d,肾小管以坏死为主,高龄鼠比低龄鼠明显(P<0.05),p53(-/-)鼠比p53(+/+)鼠明显(P<0.05);细胞凋亡在7d时达到高峰,且高龄鼠比低龄鼠明显(P<0.05),p53(+/+)鼠比p53(-/-)鼠明显(P<0.05)。p53(+/+)低龄鼠IRI后1月出现衰老细胞,3月和6月显著增多(P<0.05),对侧肾未见衰老细胞;而p53(-/-)低龄鼠IRI后6月才出现明显的衰老细胞;两种基因型高龄鼠在IRI后0d双肾均见衰老细胞,但p53(+/+)鼠显著多于p53(-/-)鼠(P<0.05),1d后,IRI肾的衰老细胞明显减少(P<0.05),对侧肾无变化。p53(+/+)高龄和低龄鼠细胞增殖无统计学意义(P>0.05),而p53(-/-)低龄和高龄鼠在IRI后3、7d细胞增殖进行性增加,且低龄鼠显著多于高龄鼠、p53(-/-)鼠多于p53(+/+)鼠(P<0.05)。对高龄鼠IRI后1d点细胞衰老和凋亡进行相关分析显示,在p53(+/+)鼠二者显著负相关(r=-0.82,P<0.05),在p53(-/-)鼠,二者无显著相关性(r=0.26,P>0.05)。结论:①IRI可促进正常肾小管上皮细胞衰老的进程;②已经进入衰老状态的肾小管上皮细胞在遭受IRI刺激后,更易走向死亡(坏死和/或凋亡);③p53在IRI所致肾小管上皮细胞演变过程中发挥着重要的调控作用。
OBJECTIVE: To observe the changes of renal tubular epithelial cells in young and old p53 (+ / +) and p53 (- / -) mice after ischemia / reperfusion injury (IRI) Effects of cell evolution. Methods: Left kidney (2 months old) and old (12 months) p53 (+ / +) and p53 (- / -) mice were established IRI model of left kidney. The histological changes of renal tubules were observed at 0d, 1d, 3d, 7d, 1 month, 3 months and 6 months after IRI. The proliferating cell nuclear antigen (PCNA) of renal tubular epithelial cells was detected by immunohistochemistry The expression of SA-β-gal in renal tubular epithelial cells was detected by histochemical staining and the expression of terminal deoxynu-cleotidyltransferase -mediated dUTP-biotininsitunick -endlabe-ling, TUNEL) assay was used to detect the apoptosis of renal tubular epithelial cells. Results: At the 0th day after IRI, the renal tubules were mainly necrotic, the aged rats were significantly better than the younger rats (P <0.05), and the p53 (- / - (P <0.05). The p53 (+ / +) mice were more obvious than the p53 (- / -) mice at the 7th day (P <0.05). The senescent cells appeared in the first month after IRI in the p53 (+ / +) young mice, which were significantly increased in March and June (P <0.05) and no senescent cells in the contralateral kidney. However, the IRI of p53 (+ / + (P <0.05). However, p53 (+ / +) mice were significantly more than p53 (- / -) mice (P <0.05) after 1 day , IRI renal aging cells was significantly reduced (P <0.05), contralateral kidney no change. The proliferation of p53 (+ / +) old and young mice was not statistically significant (P> 0.05), while the p53 (- / -) young mice and old mice increased significantly at 3 and 7 days after IRI In aged mice, p53 (- / -) mice were more than p53 (+ / +) mice (P <0.05). Correlation analysis of the cell senescence and apoptosis on the 1st day after IRI showed that there was a significant negative correlation (r = -0.82, P <0.05) between p53 (+ / +) and p53 (- / - There was no significant correlation between them (r = 0.26, P> 0.05). Conclusion: ①IRI can promote the process of normal renal tubular epithelial cell senescence; ② The tubular epithelial cells which have entered the aging state are more likely to die (necrosis and / or apoptosis) after being subjected to IRI stimulation; Cells play an important regulatory role in the evolution.