安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
Acta Universitatis Medicinalis Anhui
2015年
11期
1606-1609,1610
,共5页
陈培杰%李俊%黄成%孟晓明%蔡双朋
陳培傑%李俊%黃成%孟曉明%蔡雙朋
진배걸%리준%황성%맹효명%채쌍붕
PICK1%肝纤维化%肝星状细胞
PICK1%肝纖維化%肝星狀細胞
PICK1%간섬유화%간성상세포
PICK1%Liver Fibrosis%hepatic stellate cell
目的:研究蛋白激酶 C 结合蛋白1(PICK1)在小鼠肝纤维及活化的肝星状细胞中的表达变化,并探讨其对人肝星状细胞株(LX-2)活化的影响。方法建立四氯化碳(CCl4)诱导的小鼠肝纤维化模型,观察 PICK1在肝纤维化过程中的表达变化;转化生长因子β1(TGF-β1)刺激 LX-2活化,West-ern blot 测定 PICK1的蛋白表达情况;转染 PICK1过表达质粒至 LX-2中,再以 TGF-β1诱导活化,Western blot 检测PICK1、α平滑肌肌动蛋白(α-SMA)、I 型胶原(Col1a1)和Smad2、3及其磷酸化水平的蛋白表达情况。结果正常组小鼠肝脏中 PICK1表达较高,随着肝纤维化的加重,PICK1的表达逐渐递减。TGF-β1诱导活化的 LX-2细胞中 PICK1表达降低。转染 PICK1过表达质粒后,TGF-β1诱导的α-SMA、Colla1的表达明显减少,且 Smad2、3磷酸化水平显著下降。结论 PICK1在纤维化肝组织及活化的 HSC 中表达下调。过表达 PICK1可抑制 TGF-β1诱导的 LX-2活化,可能是通过抑制 TGF-β/Smad 通路而发挥作用,为肝纤维化的防治研究提供了新的思路和靶点。
目的:研究蛋白激酶 C 結閤蛋白1(PICK1)在小鼠肝纖維及活化的肝星狀細胞中的錶達變化,併探討其對人肝星狀細胞株(LX-2)活化的影響。方法建立四氯化碳(CCl4)誘導的小鼠肝纖維化模型,觀察 PICK1在肝纖維化過程中的錶達變化;轉化生長因子β1(TGF-β1)刺激 LX-2活化,West-ern blot 測定 PICK1的蛋白錶達情況;轉染 PICK1過錶達質粒至 LX-2中,再以 TGF-β1誘導活化,Western blot 檢測PICK1、α平滑肌肌動蛋白(α-SMA)、I 型膠原(Col1a1)和Smad2、3及其燐痠化水平的蛋白錶達情況。結果正常組小鼠肝髒中 PICK1錶達較高,隨著肝纖維化的加重,PICK1的錶達逐漸遞減。TGF-β1誘導活化的 LX-2細胞中 PICK1錶達降低。轉染 PICK1過錶達質粒後,TGF-β1誘導的α-SMA、Colla1的錶達明顯減少,且 Smad2、3燐痠化水平顯著下降。結論 PICK1在纖維化肝組織及活化的 HSC 中錶達下調。過錶達 PICK1可抑製 TGF-β1誘導的 LX-2活化,可能是通過抑製 TGF-β/Smad 通路而髮揮作用,為肝纖維化的防治研究提供瞭新的思路和靶點。
목적:연구단백격매 C 결합단백1(PICK1)재소서간섬유급활화적간성상세포중적표체변화,병탐토기대인간성상세포주(LX-2)활화적영향。방법건립사록화탄(CCl4)유도적소서간섬유화모형,관찰 PICK1재간섬유화과정중적표체변화;전화생장인자β1(TGF-β1)자격 LX-2활화,West-ern blot 측정 PICK1적단백표체정황;전염 PICK1과표체질립지 LX-2중,재이 TGF-β1유도활화,Western blot 검측PICK1、α평활기기동단백(α-SMA)、I 형효원(Col1a1)화Smad2、3급기린산화수평적단백표체정황。결과정상조소서간장중 PICK1표체교고,수착간섬유화적가중,PICK1적표체축점체감。TGF-β1유도활화적 LX-2세포중 PICK1표체강저。전염 PICK1과표체질립후,TGF-β1유도적α-SMA、Colla1적표체명현감소,차 Smad2、3린산화수평현저하강。결론 PICK1재섬유화간조직급활화적 HSC 중표체하조。과표체 PICK1가억제 TGF-β1유도적 LX-2활화,가능시통과억제 TGF-β/Smad 통로이발휘작용,위간섬유화적방치연구제공료신적사로화파점。
Objective To explore the expression of protein interacting with Ca-kinase-1(PICK1)in fibrotic liver of mice and activated hepatic stellate cell,and investigate the effect of PICK1 on the activation of hepatic stellate cell strain (LX-2).Methods Liver fibrosis model in mice was induced by CCl4 and then the PICK1 level was detec-ted in fibrotic liver tissue.LX-2 cells were activated by transforming growth factor β1 (TGF-β1),and the protein level of PICK1 was detected by Western blot.After transfected with the plasmid expressing PICK1,LX-2 was stim-ulated with TGF-β1,the levels of α-smooth muscle actin(α-SMA),collagen type I (Col1a1 ),Smad2,3 and phosphorylation level of them were determined by Western blot.Results PICK1 was highly expressed in normal liver tissues and progressively down-regulated as liver fibrosis progressed.The expression of PICK1 was down-regu-lated in activated LX-2 induced by TGF-β1.The hepatic stellate cell transfected with PICK1-plasmid showed re-markablely decreased TGF-β1-induced α-SMA and Col1a1 expression and obviously declined phosphorylation levels of Smad2 and Smad3.Conclusion The expression of PICK1 decreases in fibrotic livers and activated hepatic stel-late cell.Over-expression of PICK1 can suppress the activation of hepatic stellate cell induced by TGF-β1,and probably because of the inhibitory effect on TGF-β/Smad pathway,which provides new ideas and targets for the prevention and treatment of liver fibrosis.