去甲斑蝥素对LPS诱导的体外培养的肝细胞损伤的保护作用

Effects of norcantharidin on lipopolysaccharide-induced hepatocyte injury in vitro

目的:通过体外观察去甲斑蝥素(NCTD)对LPS所致肝细胞损伤和TNF-α表达的影响,探讨NCTD的作用及其机制。方法:胶原酶Ⅳ灌流分离培养大鼠肝细胞,将细胞分为对照组、LPS组、NCTD组。对照组用无血清DMEM培养,LPS组用LPS(40 mg/L)诱导,NCTD组用不同浓度NCTD(0.5,1.0,2.5μg/mL)与LPS(40 mg/L)共同作用,各组作用时间均为24 h。MTT法检测肝细胞的增殖情况、测定培养上清液乳酸脱氢酶(LDH)含量,ELISA法检测TNF-α和IL-6表达。结果:LPS(40 mg/L)作用于原代培养大鼠肝细胞24 h后,与对照组相比,细胞生长抑制率达27%,培养上清液LDH含量增加20倍,TNF-α和IL-6的表达以及NF-κB DNA结合活性均明显增加(P<0.05)。给予不同浓度NCTD后,培养上清液LDH,TNF-α和IL-6的含量及NF-κB DNA结合活性均明显下降,且呈量效关系。结论:NCTD拮抗LPS所致的肝细胞损伤,其保护作用的机制可能与其抑制TNF-α和IL-6的表达有关。

Objective： To study the effects ofnorcantharidin （NCTD） on lipopolysaccharide （LVS）-induced hepatocyte injury and the expression of TNF-α and IL-6 in vitro. Results： 40 mg/L LPS caused a 27% growth inhibition in primary hepatocytes. LDH leakage was 20- fold higher in NCTD-treated hepatocytes than in normal ones. TNF-α and IL-6 expression significantly increased. In cells treated with NCTD at doses of 0.5, 1.0 and 2.5 Fg/mL, LDH leakage, TNF-α and IL-6 expression, and NF-κB DNA binding activity were attenuated in a dose dependent manner. Conclusion： NCTD protects hepatocytes from in）ury induced by LPS; the protection is associated with suppression of the inflammatory cytokine TNF-α and IL-6.