内毒素诱导的库普弗细胞活化对肝细胞损伤的影响及姜黄素的干预作用

目的观察脂多糖(LPS)激活库普弗细胞(KC)后对肝细胞损伤的影响及姜黄素的干预作用。方法①分离大鼠KC,培养48h后分别添加不同浓度的姜黄素培养3h,检测乳酸脱氢酶(LDH)分析药物毒性剂量;②取无细胞毒性的3种浓度姜黄素(10mmol/L～30mmol/L)分别温育KC,另设正常组和模型对照组,1h后加LPS(浓度0.1mg/L)作用2h,然后取上清以ELISA法测定TNF-α含量;③将上述各组KC换液洗涤后,继续培养12h,然后取其培养液分别作用于分离培养24h的肝细胞,检测各时间点(作用后3h,6h,12h,24h)肝细胞上清的ALT、LDH活性。结果①姜黄素低于30mmol浓度对KC无细胞毒性;②姜黄素能显著抑制LPS刺激后KC的TNF-α的分泌量,量效关系显著,其中30mmol组接近正常水平;③经LPS作用的KC培养液对正常肝细胞有明显的损伤作用,而经姜黄素保护的各组培养液,对肝细胞的损伤作用明显减轻,呈显著的浓度依赖性。结论姜黄素可以通过抑制LPS诱导的KC活化而减轻肝细胞损伤。

Impaired Effect on Hepatocyte by LPS-induced Kupffer Cells Activation and Intervention of Curcumin

Objective To study effect of Curcumin on hepatocyte injury challenged by activated Kupffer cells induced by lipopolysaccharide(LPS).Method Kupffer cells(KC)and hepatocytes were isolated by in situ perfusion with collagenase and Nycodenz density gradient centrifugation,the isolated Kupffer cells were cultured for 48 hours,then culture supernatant was added to hepatocytes cultured for 24 hours after isolation,ALT and LDH activity in hepatocytes culture supernatant was determined after 3,6,12,24 hours treatment.Result KCs conditioned culture supernatant KCM obtained after LPS treatment showed markedly time-dependent injury to normal hepatocytes,while KCs conditioned culture supernatant treated with Curcumin showed marked dose dependent alleviated injury to hepatocytes.