CAJ | 학술논문

目的观察肝卵圆细胞(hepatic oval cell,HOC)对肝纤维化(hepatofibrosis,HF)大鼠肝组织中细胞外信号调节蛋白(extracellular regulated protein kinases,ERK)和丝裂原激活蛋白激酶(mitogen activated protein kinase,P38MAPK)信号通路蛋白的影响.方法 SD大鼠以高脂低蛋白饲养、饮用10%乙醇,皮下注射40%CCl4,隔4 d注射一次,连续8周制备HF模型.HF大鼠采用胶原酶原位灌注法分离,percall纯化原代HOC.HOC悬液0.5 ml(1×109个细胞)经门静脉植入8周HF大鼠的肝脏内,分别于8、15、30d处死大鼠,HE和Masson染色观察HF病理组织学变化,Western blot 法检测肝组织ERK与P38MAPK信号通路蛋白的表达,同时检测ALB、FGF-3、c-kit、HNF-1α、PCNA表达.结果病理组织学显示,HOC处理组大鼠HF被部分逆转,肝组织胶原纤维增生程度明显减轻,肝组织Ras、ERK、p-ERK、c-fos、c-jun、STAT3、ALB、FGF-3、PCNA蛋白表达显著下降(分别F=91.88,36.28,54.66,93.07,64.76,58.49,52.63,20.45,27.03,均P＜0.05),而HNF-α1和c-kit表达上调(分别F=18.63,25.99,均P＜0.05).结论 HOC抑制HF活化的ERK信号通路而改善肝纤维化程度,在HOC存在条件下p-P38并不激活c-fos、c-jun、STAT3、5表达,c-kit和HNF-1α表达增加,而肝组织结构损害程度明显减轻,肝纤维化程度明显改善.
목적 관찰간란원세포(hepatic oval cell,HOC)대간섬유화(hepatofibrosis,HF)대서간조직중세포외신호조절단백(extracellular regulated protein kinases,ERK)화사렬원격활단백격매(mitogen activated protein kinase,P38MAPK)신호통로단백적영향.방법 SD대서이고지저단백사양、음용10%을순,피하주사40%CCl4,격4 d주사일차,련속8주제비HF모형.HF대서채용효원매원위관주법분리,percall순화원대HOC.HOC현액0.5 ml(1×109개세포)경문정맥식입8주HF대서적간장내,분별우8、15、30d처사대서,HE화Masson염색관찰HF병리조직학변화,Western blot 법검측간조직ERK여P38MAPK신호통로단백적표체,동시검측ALB、FGF-3、c-kit、HNF-1α、PCNA표체.결과 병리조직학현시,HOC처리조대서HF피부분역전,간조직효원섬유증생정도명현감경,간조직Ras、ERK、p-ERK、c-fos、c-jun、STAT3、ALB、FGF-3、PCNA단백표체현저하강(분별F=91.88,36.28,54.66,93.07,64.76,58.49,52.63,20.45,27.03,균P＜0.05),이HNF-α1화c-kit표체상조(분별F=18.63,25.99,균P＜0.05).결론 HOC억제HF활화적ERK신호통로이개선간섬유화정도,재HOC존재조건하p-P38병불격활c-fos、c-jun、STAT3、5표체,c-kit화HNF-1α표체증가,이간조직결구손해정도명현감경,간섬유화정도명현개선.
Objective To observe the influence of hepatic oval cell (HOC) on the expression ERK and P38MAPK signaling pathway protein in liver tissue of murine experimental hepatofibrosis (HF).Method SD rats were fed with 10% ethanol and food with high-fat and low-protein, and were injected subcutaneously with carbontetrachloride once every four days for 8 weeks to establish hepatic fibrosis. HOGs were isolated from male HF rats by collagenase porfusion of the liver. HF rats at 8th week were transplanted with 0. 5 ml HOC suspension medium at a density of 1 × 109 cell /ml via portal vein, and the rats were sacrificed at 8th, 15th, 30th day respectively. Histopathologic changes of liver tissues were observed by HE and Masson. The expression of ERK and P38MAPK signaling pathway protein were determined by Western blotting. Result Hepatofibrosis was reversed and the degree of hyperplasia fibrilcollagen in hepatic fibrosis rats decreased significantly by HOC transplantion. HOC down-regulated the protein expression of Ras, ERK,p-ERK, c-fos, c-jun, STAT3, ALB, FGF-3, PCNA ( F = 91.88,36.28,54.66,93.07,64.76,58.49,52.63,20.45 ,27.03, all P ＜ 0.05 ), up-regulated the protein expression level of HNF-α1, PDGF-Rβ significantly in liver tissues(F = 18.63,25.99,P ＜0.05). Conclusions HOC improves the degree of hepatofibrosis through inhibiting hyperplasia of collagen fibril in liver tissue of hepatofibrosis rats. With the presence of HOC the expression of c-fos,c-jun,STAT3,5 was not activated by p-P38MAPK. The expression of c-kit and HNF-1α increased and that liver tissue injury alleviated, and hepatofibrosis was improved.