CXCR1/2拮抗剂G31P对大剂量顺铂所致小鼠急性肝损伤的保护作用

Protective effect of CXCR1/2 antagonist G31P against acute liver injury induced by high dose cisplatin in mice

目的利用趋化因子受体拮抗剂G31P探讨CXC趋化因子在大剂量顺铂(DDP)所致小鼠急性肝损伤中的作用及其可能的作用机制。方法利用大剂量DDP(15 mg/kg体质量)诱导小鼠急性肾功能衰竭的动物模型,C57BL/6小鼠被随机分为DDP用药组、DDP+G31P用药组(G31P用药组)和生理盐水(NS)对照组。15 mg/kg体质量DDP单次腹腔内注射,500μg/kg体质量G31P每日皮下注射1次,连用3 d,等量NS对照。分别取每组小鼠各10只,观察其用药后的反应及生存时间。取对照组小鼠及DDP用药后3、6、24、48 h的DDP用药组和G31P用药组小鼠各10只,称重后内眦静脉取血检测肝、肾功能。剖腹取肝组织,制备10%肝组织匀浆检测其中髓过氧化物酶(MPO)及白细胞介素6(IL-6)的含量,HE染色进行肝组织形态学观察。结果 DDP用药后1周内,70%的DDP用药组小鼠死亡,而G31P用药组小鼠全部存活。DDP用药后48 h小鼠出现急性肾功能衰竭,G31P可有效预防DDP所致的小鼠肾损害。DDP用药3～6 h后,小鼠外周血谷丙转氨酶(ALT)增高,并出现肝细胞变性、灶状坏死及炎细胞浸润;G31P能有效预防大剂量DDP所致的肝细胞坏死及炎细胞浸润。此外,DDP用药后6 h,G31P用药组小鼠肝组织MPO及IL-6的含量显著低于DDP用药组小鼠(P<0.01)。结论 G31P通过抑制中性粒细胞在肝组织内浸润及MPO和IL-6在肝组织内的释放从而对DDP所致的肝损害有保护作用,提示CXC趋化因子在大剂量DDP所致肝肾损害中发挥重要作用。

Objective To explore the role and possible mechanisms of CXC chemokine in acute liver injury induced by high dose cisplatin （DDP） in mice using chemokine receptor antagonist G31P. Methods Animal model of acute renal failure induced by high dose DDP （1.5 mg/kg） was used in this experiment. C57BL/6 mice were randomly divided into DDP group, DDP ＋ G31P group （G31P group） and natural saline （NS） control group. Single dose of DDP （15 mg/kg） was injected in- traperitoneally, 500 μg/kg G31P was administrated subcutaneously once daily for 3 d and the same volume of NS was given as control. Toxic effects and survival time were observed in 10 mice from each group subsequently. Blood samples were ob- tained from the orbital venous sinus of 10 mice in DDP group and G31P group before and 3, 6, 24, 48 h after DDP adminis- tration to measure blood urea nitrogen （BUN）, creatinine （Cre） and alanine aminotransferase （ALT）. The liver tissues were harvested and the morphological changes of liver tissue were observed by light microscope. The levels of peroxidase （MPO） and interleukin-6 （IL-6） in 10% liver tissue homogenate were determined by colorimetry. Results 70% mice in DDP group died within 1 week after DDP injection; while all mice in G31P group were alive. The mice suffered from acute renal failure at 48 h after DDP administration, and G3I P could protect mice from acute nephrotoxicity. Three to six hours after DDP administration, serum ALT activity in mice of DDP group increased, and liver cell degeneration, spotty necrosis and inflammatorycell infiltration could be seen in livers of mice in DDP group. G3IP could prevent liver cells from spotty necrosis and inflamma- tory cell infiltration in mice induced by DDP. The contents of MPO and IL-6 in 10% liver tissue homogenate of mice in G31P group were significantly lower than those in DDP group at 6 h after DDP administration （ P 〈0.01）. Conclusion G31P could effectively prevent liver cells from spotty necrosis and neutrophils infiltration and reduce the release of MPO and IL-6 in liver tissue of mice induced by high dose DDP, indicating CXC chemokine played an important role in high dose DDP induced acute hepato-nephrotoxicity.