摘要
目的探讨褪黑素（MT）对百草枯（PQ）染毒小鼠肝损伤的保护作用及其可能机制。方法将48只雄性BALB／c小鼠按随机数字表法分为正常组（n=6）、MT对照组（n=6）、PQ染毒组（n=18）、MT干预组（n=18，PQ染毒1h＋MT）；腹腔注射给药，MT为15mg／kg，PQ为30mg／kg；后两组再按给药后不同时间点分为12、24、72h亚组，每个亚组6只。干预后不同时间点于小鼠内眦取血并取肝组织，用碘比色法检测血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）含量，用双抗夹心酶联免疫吸附试验（ELISA）测定肝组织肿瘤坏死因子-α（TNF-α）、白细胞介素-1B（IL-1β）水平，用蛋白质免疫印迹试验（Western Blot）检测肝细胞核内核转录因子-κB p65（NF-κB p65）的蛋白表达；经苏木素-伊红（HE）染色后光镜下观察肝组织病理学改变。结果与正常组比较，PQ染毒组在染毒后12h血清ALT、AST水平和肝组织TNF-α水平即明显升高，24h达峰值[ALT（U／L）：417．88±76．16比41．76±3．63，AST（U／L）：469．79±69．81比53．19±6．31，TNF—α（pg／mg）：46．39±9．81比13．01±3．19，均P〈0．05]，之后逐渐下降；肝组织IL-1β水平和细胞核内NF-κBp65表达随时间延长持续升高，72h达峰值[IL-1β（pg／mg）：30．74±5．81比3．81±0．71，NF-κB p65蛋白（灰度值）：1．70±0．14比0．85±0．08，均P〈0．05]。MT干预可明显抑制PQ染毒导致的上述指标异常升高，与PQ染毒组比较，MT干预组血清AST和肝组织TNF—α、IL-1β水平于12h起即明显降低[AST（U／L）：269．35±11．34比391．11±8．71，TNF-α（pg／mg）：15．10±5．03比28．77±5．96，IL-1β（pg／mg）：6．23±1．03比10．89±3．02，均P〈0．05]，血清ALT水平和细胞核内NF-κB p65蛋白表达于24h起明显降低[ALT（U／L）：249．38±21．71比417．88±76．16，NF-κB p65蛋白表达（灰度值）：1．13±0．07比1．45±
Objective To observe the protective effect of melatonin （MT） on paraquat （PQ）-induced acute liver injury in mice and its possible mechanism. Methods A total of 48 male BALB/c mice were randomly divided into normal group （n=6）, MT control group （n = 6）, PQ poisoning group （n = 18）, and MT treatment group （n = 18, 1 hour-PQ poisoning ± MT）. The drugs were intraperitoneally injected, MT 15 mg/kg, PQ 30 mg/kg. The mice in the later two groups were subdivided into 12, 24, 72 hours subgroups according to different time points after administration, with 6 mice in each subgroup. Blood from medial angle of eye and liver tissue were collected at different time points after intervention to determine the serum levels of alanine transaminase （ALT） and aspartate aminotransferase （AST） with iodine colorimetry. The levels of tumor necrosis factor-α （TNF-α ） and interleukin-1β （IL-1β ） in liver tissue were determined with double-antibody sandwich enzyme linked immunosorbent assay （ELISA）. The protein expression of nuclear factor-κB p65 （NF-κB p65） in liver nuclei was determined by Western Blot. The pathological Changes in liver tissue were observed with light microscope using hematoxylin and eosin （HE） staining. Results Compared with the normal group, the levels of serum ALT and AST, and TNF-α in liver tissue were significantly increased from 12 hours after poisoning, and peaked at 24 hours [ALT （U/L）： 417.88±76.16 vs. 41.76±3.63, AST （U/L）： 469.79±69.81 vs. 53.19±6.31, TNF-α （pg/mg）： 46.39±9.81 vs. 13.01 ±3.19, all P 〈 0.05], then they were gradually decreased; the levels of IL-1β in liver tissue and NF-κB p65 in nucleus were continuously rose with time prolongation, peaked at 72 hours [IL-1β （pg/mg）： 30.74±5.81 vs. 3.81±0.71, NF-κB p65 protein （gray value）： 1.70±0.14 vs. 0.85±0.08, both P 〈 0.05]. MT intervention could significantly inhibit the above parameters with abnormal increase induced by PQ poisoning. Compared wit
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2016年第4期324-328,共5页
Chinese Critical Care Medicine
基金
浙江省温州市科技计划项目（Y20120094）
浙江省医学扶植重点建设学科项目（07-F04）
浙江省医学创新学科项目（11-CX26）
浙江省中医药重点学科项目（2012-XK-A28）
