甘草酸二铵对急性肝衰竭大鼠肝细胞凋亡的影响

Effects of glycyrrhizinate on apoptosis of hepatocytes in rat acute liver failure model

目的观察甘草酸二铵对急性肝衰竭大鼠肝细胞凋亡的影响。方法将60只健康雌性Wistar大鼠随机分为实验组、对照组、空白对照组，每组20只，实验组大鼠给予天晴甘平灌胃，对照组和空白对照组给予生理盐水灌胃；3h后实验组、对照组腹腔注射D-氨基半乳糖（D—Gal）和脂多糖（LPS）制作急性肝衰竭模型，空白对照组腹腔注射生理盐水；注射后分别于不同时间点采用酶联免疫吸附试验（ELISA）和原位缺口末端标记法（TUNEL）法检测大鼠血清肿瘤坏死因子-α（TNF-α）浓度和细胞凋亡水平。结果实验组和对照组大鼠肝组织分别于注射后6h和4h出现TUNEL阳性细胞，凋亡指数均于注射后12h达高峰，实验组为（5．23±0．28）％，对照组为（29．46±0．97）％，实验组的凋亡指数显著降低，两组间差异有统计学意义（P〈0．05）；对照组和空白对照组比较凋亡指数差异无统计学意义（P〉0．05）。实验组和对照组的血清TNF-α浓度均于注射后2h开始升高，且于注射后8h同步达高峰，实验组为（510．78±93．68）μg/L，对照组为（532．53±89．77）μg/L，两者间差异无统计学意义（P〉0．05）；对照组和空白对照组比较TNF-α差异无统计学意义（P〉0．05）。结论甘草酸二铵具有减轻急性肝衰竭大鼠肝细胞凋亡的作用，这种作用并非是通过降低血清的TNF-α浓度而实现的。

Objective To explore the effects of glycyrrhizinate on apoptosis of hepatocytes in the rats with acute liver failure. Methods Sixty healthy female Wistar rats were randomly divided into 3 groups： experimental group, control group, and normal control group （n =20 in each group）. The rats were injected intraperitoneally with D-galactosamine （D-Gal） and lipopolysaeeharide （LPS） to create the model of acute liver failure. Three h prior to the injection, the rats in experimental group were intragastrically administered diammonium glyeyrrhizinate enteric-coated capsules. Enzyme linked immunosorbent assay （ELISA） and TdT-mediated dUTP nick end labeling （TUNEL） were carried out to detect serum tumor necrosis factor-α （TNF-α） and apoptotic cells of the rats at different time points after injection respectively. Results In experiment and control groups, TUNEL-positive cells appeared at 4 h and 6 h after injection respectively, and reached a peak at 12 h synchronously. The apoptotic index in the experiment group was （5.23 ± 0. 28 ） %, significantly lower than in the control group [ （29.46 ± 0. 97 ） % ] （ P 〈 0. 05 ）. In experiment and control groups, the serum TNF-α was increased at 2 h and reached a peak at 8 h after injection synchronously. The serum TNF-α in the experiment group was （510. 78 ± 93.68 ） p.g/L, significantly different from that in the control group [ （532. 53 ± 89. 77 ） μg/L ] （ P 〉 0. 05 ）. Conclusion Glycyrrhizinate enteric-coated capsules can reduce apoptosis of hepatocytes in rats with acute liver failure, which is not achieved by reducing the concentrations of TNF-α in serum.