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目的研究姜黄素对乙醇诱导的大鼠原代肝细胞氧化损伤的防护作用。方法以二步胶原酶技术分离的SD大鼠原代肝细胞经不同剂量(0～200mmol/L)和不同时间(0～24h)暴露于乙醇,确定乙醇对肝细胞损伤的敏感的暴露时间与剂量。乙醇暴露前用姜黄素进行不同暴露剂量(0～50μmol/L)和暴露时间(0～5h)的预暴露。检测细胞培养上清液中天门冬氨酸氨基转移酶(AST)和乳酸脱氢酶(LDH)活力及肝细胞的氧化-抗氧化指标[谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)]来观察姜黄素对肝细胞免受乙醇氧化损伤的保护效应。结果与对照组相比,乙醇暴露组随着乙醇暴露浓度的增加和时间的增长,肝细胞培养液上清液中的LDH、AST活力和MDA水平升高,GSH含量和SOD活力下降。以100mmol/L暴露8h最为明显。与乙醇100mmol/L暴露8h组相比,姜黄素预暴露组(0～50μmol/L)随姜黄素剂量的增加,LDH、AST活力下降(P<0.05),姜黄素预暴露剂量为15和30μmol/L时接近对照组水平;MDA生成明显减少(P<0.05);但SOD、GSH水平明显升高(P<0.05),姜黄素预暴露剂量为15和30μmol/L时,SOD基本回复到对照组水平。15μmol/L姜黄素预暴露1和5h,各个指标与乙醇组相比,差异均有统计学意义,1h干预效果优于5h。结论姜黄素可能通过降低GSH消耗,提高抗氧化酶活力,抑制脂质过氧化来保护大鼠原代肝细胞的酒精性氧化损伤,保护作用与姜黄素剂量及暴露时间有关。
Objective To study the protective effect of curcumin on ethanol-induced oxidative damage in rat primary hepatocytes. Methods Primary hepatocytes from SD rats isolated by two-step collagenase technique were exposed to ethanol at different doses (0-200 mmol / L) and different times (0-24 h) to determine the sensitive exposure time of hepatocytes to ethanol dose. Pre-exposure with curcumin at various exposure doses (0-50 μmol / L) and exposure time (0-5 h) before ethanol exposure. The activities of aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) and the oxidative-anti-oxidative indicators of hepatic cells [glutathione (GSH), malondialdehyde (MDA) , Superoxide dismutase (SOD)] to observe the protective effects of curcumin on hepatocytes from ethanol oxidative damage. Results Compared with the control group, with the increase of ethanol exposure and the increase of the time, the activities of LDH, AST and MDA in hepatocyte culture supernatant increased, and the content of GSH and the activity of SOD decreased. The most obvious exposure to 100mmol / L for 8h. Curcumin pre-exposure group (0-50 micromol / L) decreased the activity of LDH and AST with the increase of curcumin dose (P <0.05), and the curcumin pre-exposure dose was 15 and 30 micromol / L, the level of MDA was significantly decreased (P <0.05), but the levels of SOD and GSH were significantly increased (P <0.05). When the curcumin pre-exposure dose was 15 and 30 μmol / L, SOD basically returned to the control Group level. Curcumin 15 micromol / L pre-exposure 1 and 5h, each index compared with the ethanol group, the difference was statistically significant, 1h intervention better than 5h. Conclusion Curcumin may protect alcohol oxidative damage of rat primary hepatocytes by reducing GSH consumption, increasing antioxidant enzyme activity and inhibiting lipid peroxidation. The protective effect of curcumin may be related to the dosage of curcumin and the exposure time.