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目的:研究新型重组人肿瘤坏死因子(rhTNF-NC)对培养大鼠肝细胞内游离Ca2+ 浓度([Ca2+ ]i)及小鼠肝组织中Ca2+ 含量的影响。方法:以Fura-2/AM 为Ca2+ 荧光检测,荧光分光光度法测定培养大鼠肝细胞内[Ca2+ ]i;原子吸收分光光度法测定小鼠肝组织中Ca2+ 含量。结果:在加入rhTNF-NC3 h 后,可剂量依赖性地引起培养大鼠肝细胞内[Ca2+ ]i 显著升高;rhTNF-NC可使D-GalN 致敏小鼠出现严重肝损伤,肝组织中Ca2+ 含量明显增加,钙通道阻滞剂维拉帕米、硝苯地平能显著降低肝组织中Ca2+ 含量,同时减轻肝损伤程度。结论:rhTNF-NC可引起培养大鼠肝细胞及小鼠肝组织中钙稳度失调,钙通道阻滞剂通过减少Ca2+ 超载而减轻rhTNF-NC的肝损伤作用
Objective: To investigate the effects of rhTNF-NC on intracellular free calcium concentration ([Ca2 +] i) and the content of Ca2 + in liver of mice. Methods: Fura-2 / AM was used to detect Ca2 + fluorescence and [Ca2 +] i was measured by fluorescence spectrophotometry. Ca2 + content in liver tissue was measured by atomic absorption spectrophotometry. Results: After adding rhTNF-NC3 h, the [Ca2 +] i in the cultured hepatocytes was significantly increased in a dose-dependent manner; rhTNF-NC could cause severe hepatic injury in the D-GalN-sensitized mice, Ca2 + content increased significantly, calcium channel blocker verapamil, nifedipine can significantly reduce the content of Ca2 + in liver tissue, and reduce the degree of liver injury. CONCLUSION: rhTNF-NC can cause imbalance of calcium in cultured rat hepatocytes and mouse liver, and calcium channel blocker can reduce hepatic injury of rhTNF-NC by reducing Ca2 + overload