委陵菜酸对小鼠急性肝损伤保护作用  

Protective effect of tormentic acid on LPS/D-GalN-induced acute liver injury in mice

作　　者：谭实美 梁春宏[1] 韦玲[1] 吕淑娟[1] 黄权芳[2] 林兴[1] 

机构地区：[1]广西医科大学药学院药理学教研室,广西南宁530021 [2]广西中医药大学第一附属医院

出　　处：《中国公共卫生》2016年第8期1054-1058,共5页Chinese Journal of Public Health

基　　金：国家自然科学基金(81260674);广西自然科学基金(2013GXNSFAA019146;2014GXNSFAA118155;2014GXNSFAA118154)

摘　　要：目的观察委陵菜酸(TA)对脂多糖(LPS)/D-半乳糖胺(D-Gal N)诱导的小鼠急性肝损伤的保护作用及其可能机制。方法雄性C57BL/6小鼠90只,随机分为对照组、模型组、阳性对照组及委陵菜酸低、中、高剂量组,各组小鼠分别灌胃给予受试物,连续3 d,末次给予受试物8 h后,除对照组外,其余小鼠腹腔注射LPS/D-Gal N,1.5 h后眼球取血,6 h后处死小鼠,取肝脏,观察肝组织损伤程度;采用试剂盒测定小鼠血清中门冬氨酸氨基转移酶(AST)、谷丙氨酸氨基转移酶(ALT)、总胆红素、肿瘤坏死因子-α(TNF-α)、一氧化氮(NO)、白细胞介素-6(IL-6)和IL-10水平;RT-PCR法检测TNF-α和诱导型一氧化氮合酶(i NOS)基因表达;ELISA法测定NF-κB p65蛋白含量;Western blot法检测细胞色素C和Bcl-2含量;另外,取肝组织匀浆检测caspase-3、8、9活性。结果与对照组比较,模型组小鼠ALT、AST、TNF-α、NO含量[分别为(1 833.5±125.4)、(2 183.5±138.9)U/L、(656.8±64.7)pg/m L、(205.28±16.79)μmol/L]明显升高(P<0.05),肝脏中NF-κB p65表达[(228.9±14.2)pg/mg]升高、Bcl-2蛋白表达(0.18±0.03)下降、Caspase-3蛋白表达(275.3±14.7)升高(P<0.05);与模型组比较,委陵菜酸3 mg/kg组小鼠血清中ALT、AST、TNF-α、NO含量[分别为(991.3±70.2)、(968.5±68.6)U/L、(226.5±35.9)pg/m L、(133.69±9.79)μmol/L]明显降低(P<0.05)、肝脏中NF-κB p65表达[(151.2±6.5)pg/mg]降低、Bcl-2蛋白表达(0.35±0.02)升高、caspase-3蛋白表达(156.3±5.9)下降(P<0.05)。结论委陵菜酸可保护LPS/D-Gal N诱导的小鼠急性肝损伤,其机制可能与抑制NF-κB信号通路减轻炎症反应、抑制凋亡蛋白表达、继而减少细胞凋亡有关。Objective To study the protective effect and mechanism of tormentic acid（ TA） on acute hepatic injury induced by lipopolysaccharides（ LPS） / D-galactosamine（ D-Gal N） in mice. Methods A total of 90 male C57 BL /6mice were divided into 6 groups（ 15 mice per group） ： normal control,model control,silymarin control,and low-,mediumand high-dose TA-treated groups. The mice were orally administered the drugs consecutively for 3 days. Eight hours after the last treatment,all the groups,with the exception of normal control,were intraperitoneally administered with LPS / DGal N. Blood and liver samples were sequentially collected 1. 5 and 6 hours after the treatment. The degree of tissue injury was observed with pathological section; serum levels of aspartate aminotransferase（ AST）,alanine aminotransferase（ ALT）,total bilirubin,tumor necrosis factor-α（ TNF-α）,nitric oxide（ NO）,interleukin-6（ IL-6）,and interleukin-10（ IL-10） were detected using commercial test kits; the expression of TNF-α and inducible nitric oxide synthase（ i NOS）genes were detected with reverse transcriptase-PCR（ RT-PCR）; the nuclear factor κB（ NF-κB） p65 content was measured using a enzyme-linked immunosorbent assay（ ELISA） kit; cytochrome C and Bcl-2 were analyzed with Western blot immunoassay; the activities of caspase-3,8 and 9 were detected using fluorescence assay. Results Compared to those of the normal control group,the levels of ALT（ 1 833. 5 ± 125. 4 U / L）,AST（ 2 183. 5 ± 138. 9 U / L）,TNF-α（ 656. 8 ± 64. 7 pg /m L）,NO（ 205. 28 ± 16. 79 μmol/L）,NF-κB p65（ 228. 9 ± 14. 2 pg /mg）,caspase-3（ 275. 3 ±14. 7） were significantly increased and Bcl-2（ 0. 18 ± 0. 03） were significantly decreased in the model control group（ P 0. 05）; compared with those of the model control group,the levels of ALT（ 991. 3 ± 70. 2 U /L）,AST（ 968. 5 ±68. 6 U / L）,TNF-α（ 226. 5 ± 35. 9 pg / m L）,NO（ 133. 69 ± 9. 79 μmol / L）,NF-κB p65（ 151. 2 ± 6

关 键 词：委陵菜酸(TA) 脂多糖/D-半乳糖胺(LPS/D-GalN) NF-ΚB信号通路 细胞凋亡 

分 类 号：R575[医药卫生—消化系统]