灯盏花素对大鼠急性肝细胞损伤的保护作用

Investigation of the protective effect of breviscapine on damage of rat acute hepatocyte

目的探讨灯盏花素对大鼠急性肝损害的保护作用。方法取Wistar大鼠18只,随机分为Ⅰ、Ⅱ、Ⅲ3组各6只。胶原酶原位灌流法分离大鼠肝细胞后于Ⅰ、Ⅱ、Ⅲ组分别加入PBS 20mmol/L、LPS 3mg/L、LPS 3mg/L+灯盏花素20mmol/L,于施加因素后2、4、6、8、12、24h分别收集肝细胞和肝细胞培养液进行各指标测定。用透射电镜观察肝细胞形态学变化,用Fura-2-AM测定细胞内游离钙离子浓度,用[3H]油酸标记的生物膜法测定分泌型磷脂酶A2(secretory phospholipase A2,sPLA2),用Elisa法检测前列腺素E2(PGE2)的活性。结果Ⅱ组凋亡细胞与坏死细胞数均多于Ⅰ、Ⅲ组,差异均有统计学意义(P<0.01)。LPS加入肝细胞培养液2h、8h,Ⅱ组大鼠肝细胞中Ca2+浓度高于Ⅰ、Ⅲ组(P<0.05);Ⅱ组sPLA2、PGE2水平均高于Ⅰ组、Ⅲ组,差异均有统计学意义(P<0.01)。结论灯盏花素能显著抑制急性肝损害,可能与其对钙的拮抗和对sPLA2的抑制作用有关。

Objective To investigate the protective effect of breviscapine on damage of rat acute hepatocyte. Methods 18 Wistar rats were randomly divided into I , II, III three groups, each of 6 rats. The rat hepatocyte were isolated by perfusion with collagenase. I , II, III group were separately added to PBS 20mmol/L, LPS 3mg/L, LPS 3mg/L ＋ breviscapine 20mmol/L. After administration 2,4,6,8,12,24 h, collected hepatocyte and hepatocyte culture medium for each index determination. Transmission electron microscope technique had been used to observe morphological change of hepatocyte. Intracellular Ca2＋ concentration was measured by Fura-2-AM. sPLA2 activity was detected by means of labeled E. coli membrane with [ 3 H ] oleic acid. Detected the activity of prostaglandin E2 （ PGE2 ） by Elisa method. Results The apoptotic cell and necrotic cell number of lI group were more than those of Ili group, the difference were statistically significant（ P 〈 0.01 ）. 2h,Sh after the LPS added to the hepatocyte culture fluid, the Ca2 ＋ concentration of rat hepatocyte in II group was higher than that in III group（ P 〈 0.05 ）. The sPLA2 .PGE2 average level of II group were higher than those of I group and In group,the difference were statistically significant（P 〈 0.01 ）. Conclusion Breviscapine significantly inhibited acute hepatocyte damage, may be related to calcium antagonistic and sPLA2 inhibition.