CAJ | 학술논문

目的探讨内质网应激抑制剂4-苯基丁酸(PBA)对四氯化碳(CCl4)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CCl412、24、48、72 h组、PBA+CCl4 12 h组、PBA+CCl4 24 h组、PBA+CCl4 48 h组、PBA+CCl4 72h组,每组6只.对CCl4及PBA+CCl4各组小鼠经腹腔注射CCl4 (300 μl/kg),PBA+CCl4各组小鼠在给予CCl4前经腹腔注射PBA (400 mg/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,Westem 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2 α(p-eIF2 α)、PCNA、磷酸化丝氨酸/苏氨酸蛋白激酶B(p-akt)及核因子-κ B p65(NF-κ B p65)蛋白水平.对数据进行t检验分析. 结果与相同时点CCl4组相比,PBA+CCl4组小鼠肝脏ALT水平有不同程度的降低,其中24h时点最明显[(7 423.81±581.75) U/L与(2 876.19±179.76) U/L,t=14.984,P＜0.01],而小鼠肝质量体质量比在48 h时点的差异存在统计学意义(0.072±0.001与0.064±0.004,t=3.915,P＜0.01);病理学及TUNEL检测结果显示,PBA处理可减轻CCl4引起的肝细胞坏死及凋亡程度;Western blot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-eIF2 α)水平(P＜0.01).进一步研究结果显示,与CCl4组相比,在相同时点PBA+CCl4组中与肝细胞增殖相关的p-akt、PCNA及NF-κB p65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CCl4组48 h及72 h时,肝细胞增殖明显,而PBA+CCl4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,但同时也抑制肝细胞增殖的速度.
목적 탐토내질망응격억제제4-분기정산(PBA)대사록화탄(CCl4)유도급성간손상적영향급기부분분자궤제. 방법 장60지성년건강웅성ICR소서수궤분위대조조、PBA조화CCl412、24、48、72 h조、PBA+CCl4 12 h조、PBA+CCl4 24 h조、PBA+CCl4 48 h조、PBA+CCl4 72h조,매조6지.대CCl4급PBA+CCl4각조소서경복강주사CCl4 (300 μl/kg),PBA+CCl4각조소서재급여CCl4전경복강주사PBA (400 mg/kg).채집각조소서혈액화간조직,검측혈청ALT수평,HE염색분석간장병이학개변,TUNEL기술검측세포조망정황,면역조직화학검측간장증식세포핵항원(PCNA)분포,Westem blot검측간장포도당조절단백78 (GRP78)、C/EBP가족동원단백(CHOP)、린산화c-Jun안기말단격매(p-JNK)、린산화진핵생물번역기시인자2 α(p-eIF2 α)、PCNA、린산화사안산/소안산단백격매B(p-akt)급핵인자-κ B p65(NF-κ B p65)단백수평.대수거진행t검험분석. 결과 여상동시점CCl4조상비,PBA+CCl4조소서간장ALT수평유불동정도적강저,기중24h시점최명현[(7 423.81±581.75) U/L여(2 876.19±179.76) U/L,t=14.984,P＜0.01],이소서간질량체질량비재48 h시점적차이존재통계학의의(0.072±0.001여0.064±0.004,t=3.915,P＜0.01);병이학급TUNEL검측결과현시,PBA처리가감경CCl4인기적간세포배사급조망정도;Western blot결과현시PBA처리가강저간장내질망응격상관단백분자(GRP78、CHOP、p-JNK、p-eIF2 α)수평(P＜0.01).진일보연구결과현시,여CCl4조상비,재상동시점PBA+CCl4조중여간세포증식상관적p-akt、PCNA급NF-κB p65단백수평명현강저(P＜ 0.01);면역조직화학방법검측PCNA결과야현시,재CCl4조48 h급72 h시,간세포증식명현,이PBA+CCl4조간세포증식유소감약. 결론 내질망응격억제제PBA감경간세포배사화조망적정도,단동시야억제간세포증식적속도.
Objective To explore the effects and the molecular mechanisms of 4-phenylbutyric acid (PBA) on carbon tetraehloride (CCl4)-induced acute liver injury in mice.Methods Sixty adult,healthy,male ICR mice were divided equally into the control group,PBA group,CCl4 12 h group,CCl4 24 h group,CCl4 48 h group,CCl4 72 h group,PBA+CCl4 12 h group,PBA+CCl4 24 h group,PBA+CCl4 48 h group,and PBA+CCl4 72 h group.The CCl4 groups and the PBA+CCl4 groups were intraperitoneally (i.p.) injected with CCl4 (300 mL/ kg).In the PBA+CCl4 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 CCl4 treatment.Serum alanine aminotransferase (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 2α subunit (p-eIF2α),phosphorylated serine threonine kinase (p-akt),and nuclear factor-kappa B p65 (NF-κ Bp65) were determined by western blot.Results The serum ALT level in the PBA+CCl4 groups was reduced as compared with that in the CCl4 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 time point (P ＜ 0.01).PBA reduced the degree of hepatic necrosis and apoptosis caused by CCl4,and reduced the expression of hepatic GRP78 and other endoplasmic reticulum stress-related proteins (P ＜ 0.01).The protein levels of p-akt,NF-κ Bp65 and PCNA was significantly decreased in the PBA+CCl4 groups (P ＜ 0.01).Conclusion The endoplasmic reticulum stress inhibitor PBA alleviated acute hepatic necrosis and apoptosis but restrained hepatic proliferation.