4-苯基丁酸对四氯化碳诱导小鼠急性肝损伤的影响  

Effects of 4-phenylbutyric acid on carbon tetrachloride-induced acute liver injury in mice

作　　者：齐军[1] 陈熙[1] 张程[2] 陶莉[1] 何薇[1] 王建青[4] 李俊[3] 徐德祥[2] 

机构地区：[1]安徽医科大学第一附属医院消化内科,合肥230022 [2]安徽医科大学公共卫生学院毒理学教研室 [3]安徽医科大学药学院 [4]安徽医科大学第二附属医院药剂科

出　　处：《中华肝脏病杂志》2015年第4期286-291,共6页Chinese Journal of Hepatology

基　　金：国家自然科学基金（81172711、81001480）2013年高等学校博士学科点专项（20133420110005）;安徽省自然科学基金（1308085MH120）安徽省高校省级自然科学研究重点项目（KJ201IA159）

摘　　要：目的探讨内质网应激抑制剂4-苯基丁酸（PBA）对四氯化碳（CC14）诱导急性肝损伤的影响及其部分分子机制。方法将60只成年健康雄性ICR小鼠随机分为对照组、PBA组和CC1412、24、48、72h组、PBA＋CC1412h组、PBA＋CC1424h组、PBA＋CC1448h组、PBA＋CC1472h组，每组6只。对CC14及PBA＋CC14各组小鼠经腹腔注射CC14（300μl／kg），PBA＋CC14各组小鼠在给予CC14前经腹腔注射PBA（400mg／kg）。采集各组小鼠血液和肝组织，检测血清ALT水平，HE染色分析肝脏病理学改变，TUNEL技术检测细胞凋亡情况，免疫组织化学检测肝脏增殖细胞核抗原（PCNA）分布，Western blot检测肝脏葡萄糖调节蛋白78（GRP78）、C／EBP家族同源蛋白（CHOP）、磷酸化c-Jun氨基末端激酶（p-JNK）、磷酸化真核生物翻译起始因子2a（p-eIF2a）、PCNA、磷酸化丝氨酸／苏氨酸蛋白激酶B（p-akt）及核因子-kB p65（NF-kB 965）蛋白水平。对数据进行t检验分析。结果与相同时点CC14组相比，PBA＋CC14组小鼠肝脏ALT水平有不同程度的降低，其中24h时点最明显[（7423．81±581．75）U／L与（2876．19±179．76）U／L，r=14．984，P〈0．01]，而小鼠肝质量体质量比在48h时点的差异存在统计学意义（0．072±0．001与0．064±0．004，f=3．915，P〈0．01）；病理学及TUNEL检测结果显示，PBA处理可减轻CC14引起的肝细胞坏死及凋亡程度，Westernblot结果显示PBA处理可降低肝脏内质网应激相关蛋白分子（GRP78、CHOP、p-JNK、p-eIF2a）水平（P〈0．01）。进一步研究结果显示，与CC14组相比，在相同时点PBA＋CC14组中与肝细胞增殖相关的p-akt、PCNA及NF-1cBp65蛋白水平明显降低（P〈0．01）；免疫组织化学方法检测PCNA结果也显示，在CC14组48h及72h时，肝细胞增殖明显，而PBA＋CCh组肝细胞增殖有所减弱。结论内质网应激抑制剂PBA减轻肝细胞坏死和凋亡的程度，但同时也抑制肝�Objective To explore the effects and the molecular mechanisms of 4-phenylbutyric acid （PBA） on carbon tetrachloride （CC14）-induced acute liver injury in mice. Methods Sixty adult, healthy, male ICR mice were divided equally into the control group, PBA group, CC14 12 h group, CC14 24 h group, CCL 48 h group, CCh 72 h group, PBA＋CC14 12 h group, PBA＋CC14 24 h group, PBA＋CCI4 48 h group, and PBA＋CCI4 72 h group. The CC14 groups and the PBA＋CCI4 groups were intrapefitoneally （i.p.） injected with CC14 （300 mL/kg）. In the PBA＋CCI4 groups, the mice were i.p. injected with PBA （400 mg/kg）. All mice were sacrificed to collect blood and liver specimens at different time points after the CC!4 treatment. Serum alanine aminotmusferase （ALT） was detected. Histological examination was performed using hematoxylin-eosin staining and light microscopy, and apoptosis was detected using terminal transferase dUTP nick end labeling. The hepatic distribution of proliferating cell nuclear antigen （PCNA） was detected by immunohistochemistry. The hepatic protein expression of glucose-regulated protein （GRP78）, C/EBP homologousprotein （CHOP）, phosphorylated c-Jun N-terminal kinases （p-JNK）, phosphorylated eukaryotic initiation factor 2a subunit （p-elF2a）, phosphorylated serine threonine kinase （p-akt）, and nuclear factor-kappa B p65 （NF-r, Bp65） were determined by western blot. Results The serum ALT level in the PBA＋CCI4 groups was reduced as compared with that in the CC14 groups at the various time points examined. The liver-to-body weight ratio of two groups showed a significant difference only at the 48 h lime point （P 〈 0.01）. PBA reduced the degree of hepatic necrosis and apoptosis caused by CC14, and reduced the expression of hepatic GRP78 and other endoplasrnic reticulum stress-related proteins （P〈 0.01）. The protein levels ofp-akt, NF-r, Bp65 and PCNA was significantly decreased in the PBA＋CC14 groups （P〈 0.01）. Conclusion The endoplasmic reticulum

关 键 词：细胞凋亡 内质网应激 急性肝损伤 四氯化碳 4-苯基丁酸 

分 类 号：R575[医药卫生—消化系统]