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目的:探讨高压氧对大鼠肾缺血再灌注损伤(IRI)的影响及其机制。方法:成年健康Wistar大鼠60只,随机分为4大组,正常对照组(n=6)、假手术组(n=18)、肾缺血再灌注组(IR组,n=18)和肾缺血再灌注+HBO治疗组(HBO治疗组,n=18)。后3组又按再灌注后1h、3h和5h各分为3个亚组,每个亚组大鼠6只。各组先切除右肾,取左肾研究。正常对照组切除右肾后立即切取左肾备用。假手术组不阻断左肾动脉,肾IR组和HBO治疗组则采用钳夹肾动脉法建立左肾缺血再灌注损伤模型。HBO治疗组:在肾缺血再灌注后0h、2h和4h时分别将大鼠置于动物实验用高压氧舱内治疗1h之后,治疗压力为2(Atmosphere absolute,ATA)(0.1MPa),舱内氧浓度在98%以上。各组分别于再灌注后1h、3h和5h时检测血中SCr、BUN、MDA和TNF-α的含量,并取左肾组织用于石蜡切片和HE染色、流式细胞术(Flow cytometry,FCM)行Amn-nexin V/PI双染法检测细胞凋亡以及电镜检查。结果:(1)在IR组,血中检测指标量显著高于同时点正常对照组和假手术组(P<0.05);在HBO治疗组,各指标在术后不同时点的含量均较IR组明显降低(P<0.05)。(2)FCM结果显示,HBO治疗组细胞凋亡也显著低于同期IR组(P<0.05)。(3)肾组织中细胞凋亡与血中BUN、SCr、MDA和TNF-α值呈显著正相关(P<0.05)。(4)肾组织病理和肾组织超微结构的变化在HBO治疗组的损伤也明显轻于其他组。结论:(1)肾缺血再灌注后,血中氧自由基和促炎症因子的大量产生可促进肾组织细胞凋亡,从而加重肾损伤。(2)肾缺血再灌注后,肾组织大量细胞凋亡是导致肾功能损害的重要原因。(3)HBO抑制氧自由基和促炎症因子的产生,抑制组织细胞凋亡是HBO在肾缺血再灌注损伤过程中发挥肾保护效应的重要机制。(4)早期HBO治疗对减轻肾缺血再灌注损伤进而保护肾功能更为有利。
Objective: To investigate the effect of hyperbaric oxygen on renal ischemia-reperfusion injury (IRI) in rats and its mechanism. Methods: Sixty adult healthy Wistar rats were randomly divided into 4 groups: normal control group (n = 6), sham operation group (n = 18), renal ischemia reperfusion group (n = 18) Kidney ischemia-reperfusion + HBO treatment group (HBO treatment group, n = 18). The latter 3 groups were divided into 3 subgroups according to 1h, 3h and 5h after reperfusion, and 6 rats in each subgroup. The first group removed the right kidney, left kidney. Normal control group immediately after excision of the right kidney left kidney reserve. Sham-operation group did not block the left renal artery, kidney IR group and HBO treatment group were used to establish renal artery ischemia-reperfusion injury model of renal artery clamp. HBO treatment group: Rats were placed in animal experimental hyperbaric oxygen chamber for 1 h at 0h, 2h and 4h after renal ischemia-reperfusion, respectively. The treatment pressure was 2 (ATA) (0.1MPa) The oxygen concentration is above 98%. The levels of SCr, BUN, MDA and TNF-α in the blood were measured at 1h, 3h and 5h after reperfusion, and the left kidney tissue was taken for paraffin section and HE staining. Flow cytometry (FCM ) Amn-nexin V / PI double staining was used to detect apoptosis and electron microscopy. Results: (1) In the IR group, the amount of blood test index was significantly higher than that of the normal control group and the sham operation group (P <0.05); in the HBO treatment group, Group was significantly lower (P <0.05). (2) FCM results showed that the apoptosis of HBO-treated group was also significantly lower than that of the same period IR group (P <0.05). (3) There was a significant positive correlation between apoptosis in renal tissue and BUN, SCr, MDA and TNF-α in blood (P <0.05). (4) The changes of renal histopathology and ultrastructure of renal tissue in the HBO treatment group were also significantly less than other groups. Conclusion: (1) After renal ischemia-reperfusion, a large number of production of oxygen free radicals and proinflammatory cytokines can promote renal cell apoptosis, thereby aggravating renal injury. (2) renal ischemia-reperfusion, a large number of renal cell apoptosis is an important cause of renal dysfunction. (3) HBO inhibits the production of oxygen free radicals and proinflammatory cytokines and inhibits the apoptosis of tissues is an important mechanism of renal protective effect of HBO during renal ischemia-reperfusion injury. (4) Early HBO treatment is more beneficial to reduce renal ischemia-reperfusion injury and thus renal function.