钙库操纵的钙离子通道抑制剂对百草枯致肝肾损伤的作用

The effect of store-operated calcium entry inhibitor on liver and kidney injury induced by paraquat

目的探讨钙库操纵的钙离子通道(store-operated calcium entry, SOCE)抑制剂SKF96365对百草枯(paraquat, PQ)致肝肾损伤的作用。方法体外培养A549细胞, 分为对照组(DMSO组、SKF 2 μmol/L组, SKF 10 μmol/L组), PQ组(PQ+DMSO组、PQ+SKF 2 μmol/L组, PQ+SKF 10 μmol/L组)。采用荧光素酶报告基因技术检测A549细胞活化T细胞核因子(nuclear factor of activated T cells, NFAT)激活水平变化。构建PQ中毒的小鼠模型, 分为对照组、SKF组、PQ组、PQ+SKF组。PQ组按照50 mg/kg腹腔注射PQ;SKF组按照10 mg/kg每天腹腔注射SKF96365, 共注射3 d。PQ+SKF组按照50 mg/kg腹腔注射PQ一次, 按照10 mg/kg每天腹腔注射SKF96365, 共注射3 d。第4天处死小鼠取肝肾组织, 苏木精-伊红(H&E)染色观察肝脏、肾脏组织病理学变化, TUNEL染色观察肝肾组织凋亡情况。正态分布计量资料多组间均数比较采用单因素方差分析, 两组间比较采用LSD-t检验。结果荧光素酶报告基因技术检测结果显示PQ组NFAT明显被激活, 给予SKF96365预处理后, NFAT激活明显下降, 且具有剂量依赖性。HE染色提示PQ染毒组肝脏、肾脏组织轮廓结构不清、细胞肿胀, 大量炎性细胞浸润;PQ+SKF组肝脏细胞肿胀减轻, 炎性细胞浸润明显减少。TUNEL染色提示PQ染毒组肝脏、肾脏组织凋亡细胞增加, PQ+SKF组凋亡明显减少。结论 SOCE抑制剂SKF96365能够明显减轻PQ引起的肝脏、肾脏损伤。

Objective To investigate the effect of SKF96365,store-operated calcium entry(SOCE)inhibitor,on liver and kidney injury induced by paraquat(PQ).Methods A549 cells were cultured in vitro and divided into the control group(DMSO group,SKF 2 μmol/L group,and SKF 10 μmol/L group)and PQ group(PQ+DMSO group,PQ+SKF 2μmol/L group,and PQ+SKF 10μmol/L group).The nuclear factor of activated T cells(NFAT)in A549 cells was detected by luciferase reporter gene technique.The mouse model of PQ poisoning was constructed and divided into the control group,SKF group,PQ group and PQ+SKF group.In the PQ group,mice were injected with 50 mg/kg PQ intraperitoneally;in the SKF group,mice were injected intraperitoneally with 10 mg/kg SKF96365 for 3 days.Mice in the PQ+SKF group received 50 mg/kg intraperitoneal injection of PQ once and 10 mg/kg intraperitoneal injection of SKF96365 daily for 3 days.On the fourth day,the mice were sacrificed,and the liver and kidney tissues were taken.The histopathological changes of liver and kidney tissues were observed by hematoxylin-eosin(H&E)staining,and the apoptosis of liver and kidney tissues was observed by TUNEL staining.One-way analysis of variance was used to compare the mean values of normally distributed measurement data between groups.Comparisons between groups were performed using the least significant difference Mest.Results The luciferase reporter gene technology showed that NFAT was significantly activated in the PQ group.After pretreatment with SKF96365,NFAT activation decreased sharply in a dose-dependent manner.HE staining showed that the outline structure of liver and kidney tissues in the PQ groups were unclear,cells swelled and a large number of inflammatory cells infiltrated;in the PQ+SKF group,liver cell swelling and inflammatory cell infiltration decreased significantly.TUNEL staining showed that the apoptotic cells in liver and kidney tissues increased in the PQ groups,and the apoptosis decreased remarkably in the PQ+SKF group.Conclusions SOCE inhibitor SKF96365 can significantly reduce the liver and kidney injury caused by PQ.