美他多辛对酒精性肝病大鼠肝损伤的保护作用与抑制肝组织内炎性细胞因子水平有关

doi:10.3969/j.issn.1672-5069.2016.05.014

摘要/Abstract

摘要： 目的观察美他多辛对酒精性肝病（ALD）大鼠的保护作用及其对血清细胞因子水平的影响。方法将24只雄性Wistar大鼠分为对照组（NC,n=8）、酒精性肝病组（ALD,n=8）和美他多辛治疗组（MT,n=8）。在NC组和ALD组,给予等渗盐水灌胃,给予MT组等渗盐水和美他多辛（300 mg·kg^-1·d^-1）灌胃。2 w后,继续给予NC组等渗盐水,而在ALD组和MT组,给予50%酒精5 g·kg^-1灌胃,1次/h,共3次。在末次灌胃8 h取血,采用ELISA法检测NF-κB、TNF-α、IL-1β、IL-6、IL-8和IL-10;取肝组织,采用RT-PCR法检测NF-κB和炎症因子mRNA水平。两个样本均数的比较采用t检验或近似t检验,多个样本均数的比较采用LSD检验。结果ALD组血清ALT较NC组显著升高[（100.13±10.64） U/L对（33.37±4.81） U/L,P<0.05],血清AST和GGT水平也显著升高（P值均<0.05）;正常组动物血清IL-10、IL-1β、TNF-α、IL-6、IL8和NF-ΚB水平分别为（14.73±2.03） pg/ml、（92.38±12.85） pg/ml、（2.66±0.81） pg/ml、（43.57±10.62）ng/ml、（0.29±0.07） ng/ml和（679.45±36.38） pg/ml,ALD动物分别为（16.19±1.94） pg/ml、（1927±233.69）pg/ml、（16.92±2.38） pg/ml、（127.49±9.33） ng/ml、（2.63±0.22） ng/ml和（1247.35±146.05） pg/ml,而美他多辛处理组则分别为（36.81±4.53） pg/ml、（304.13±34.79） pg/ml、（8.83±1.01）pg/ml、（81.98±8.02） ng/ml、（1.45±0.22） ng/ml和（814.84±82.40） pg/ml,提示ALD组血清TNF-α、IL-1β、IL-6和IL-8水平显著高于NC组（P值均<0.05）,MT组血清TNF-α、IL-1β、IL6和IL-8水平显著低于ALD组（P值均<0.05）,IL-10水平显著高于ALD组（P<0.05）;ALD组肝组织TNF-α、IL-1β、IL-6和IL-8 mRNA水平显著高于NC组（P值均<0.05）,MT组TNF-α、IL-1β、IL-6和IL-8 mRNA水平显著低于,而IL-10 mRNA水平显著高于ALD组（P<0.05）。结论美他多辛对酒精性肝病大鼠的肝损伤有显著的保护作用,其机制可能是抑制了与NF-κB相关的炎症反应,进而抑制了TNF-α、IL-1β、IL6和IL-8等促炎因子水平,升高了抗炎因子IL-10水平有关。

关键词: 酒精性肝病, 美他多辛, NF-κ, B, 细胞因子, 大鼠

Abstract: Objective To investigate the effect of metadoxine（MT） on acute alcoholic liver injury in rats with alcohol-induced liver diseases（ALD）. Methods Twenty-four Wistar rats were randomly divided into normal control,alcoholic liver disease and metadoxine-intervened group,with 8 rats in each group. All rats were fed with normal chow. In the first two weeks,the rats in control and in ALD groups were administered normal saline,while in MT group were treated with metadoxine at dose of 300 mg·kg^-1·d^-1. The protocol lasted for 2 weeks. Then,the rats in ALD group and MT group were made acute liver injuries by alcohol gavage with 50%（v/v） ethanol at dose of 5 g·kg^-1 once every 12 hours,and a total of 3 times,while in rats in NC group were still administered normal saline. Eight hours after the last ethanol administration,the rats were sacrificed,and the blood and liver tissues were collected. The hepatic inflammatory cytokine mRNA was assayed by RT-PCR,and serum NF-κB,TNF-ɑ,IL-1β,IL-6,IL-8 and IL-10 levels were detected by ELISA. The t-test was used for statistical analysis. Results Serum ALT levels in ALD group elevated obviously than in control [（100.13±10.64） U/L vs. （33.37±4.81） U/L,P<0.05],and serum AST and GGT levels increased also（all P<0.05）;serum IL-10, IL-1β,TNF-α,IL-6,IL8 and NF-ΚB levels in control group were（14.73±2.03） pg/ml,（92.38±12.85） pg/ml,（2.66±0.81）pg/ml,（43.57±10.62） ng/ml,（0.29±0.07） ng/ml and （679.45±36.38） pg/ml,in ALD group were （16.19±1.94）pg/ml,（1927±233.69） pg/ml,（16.92±2.38） pg/ml,（127.49±9.33） ng/ml,（2.63±0.22） ng/ml and（1247.35±146.05）pg/ml,while in MT group were （36.81±4.53） pg/ml,（304.13±34.79） pg/ml,（8.83±1.01） pg/ml,（81.98±8.02） ng/ml,（1.45±0.22） ng/ml and（814.84±82.40） pg/ml,respectively,suggesting serum TNF-α,IL-1β,IL-6 and IL-8 levels were higher than those in control（all P<0.05）,serum TNF-α,IL-1β, IL6 and IL-8 levels in MT group were lower,and serum IL-10 level in MT group was higher than in ALD group（all P<0.05）;hepatic TNF-α,IL-1β,IL-6 and IL-8 mRNA levels in ALD group were much higher than in NC group（all P<0.05）,hepatic TNF-α,IL-1β,IL-6 and IL-8 mRNA levels in MT group were much lower,while IL-10 mRNA level much higher than in ALD group（P<0.05）. Conclusion Metadoxine can protect the alcohol-induced acute liver injuries by down-regulating pro-inflammatory cytokines and alleviating hepatic inflammation.

Key words: Alcoholic liver injuries, Metadoxine, Cytokines, NF-κB, Rats

温博, 徐贝贝, 王炳元. 美他多辛对酒精性肝病大鼠肝损伤的保护作用与抑制肝组织内炎性细胞因子水平有关[J]. 实用肝脏病杂志, 2016, 19(5): 565-569.

Wen Bo, Xu Beibei, Wang Bingyuan.. Metadoxine relieving acute alcoholic liver injuries in rats with alcohol-induced liver diseases[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2016, 19(5): 565-569.

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