摘要
目的观察蛋白激酶C(PKC)活性改变对肝星状细胞(HSC)表达血小板源生长因子(PDGF)的影响及对HSC增殖和激活的作用。方法将肝星状细胞系rHSC-99随机分为3组:对照组(A组);PKC激动剂PMA 0.5μmol/L组(B组);PKC抑制剂Calphostin C 100 nmol/L组(C组)。加药后0、3、6、12、24 h分别检测各组细胞PKC活性的变化;作用24 h后,采用Western blot方法检测各组细胞PDGF和胞外信号调节激酶(ERK)的表达;采用免疫荧光化学方法检测各组细胞α-平滑肌肌动蛋白(α-SMA)的表达;采用MTT法检测细胞的增殖情况。结果PMA作用后PKC的活性显著增强,而Calphostin C则抑制PKC的活性。PKC活性增强后,与对照组比较HSC的PDGF表达升高了1.4倍(P<0.01),ERK表达升高了1.2倍(P<0.05);α-SMA的表达升高了1.3倍(P<0.01),并促进HSC的增殖;PKC活性抑制后则能抑制以上的作用。结论PKC活性的改变能调控HSC中PDGF的表达,在HSC的增殖和激活中发挥调节作用。
Objective To investigate the effect of protein kinase C （PKC） activity on the expression of platelet derived growth factor （PDGF） and proliferation of hepatic stellate cells （ HSCs）. Methods rHSC-99 HSCs were randomly divided into 3 groups. Group A served as a control. HSCs were incubated with PKC agonist PMA （0.5 μmol/L） as group B or PKC inhibitor Calphostin C （100 nmol/L） as group C. The PKC activity was detected at different incubation time points （0,3,6,12 and 24 h） ; Western blotting was used to detect the expression of PDGF and ERK and immunofluorescence was used to detect the expression of a-smooth muscle actin （α-SMA） 24 h later. Cell proliferation was assessed by MTT colorimetric assay. Results PMA increased the activity of PKC significantly,whereas Calphostin C inhibited the activity of PKC. The increased activity of PKC promoted HSC to express PDGF, ERK and α-SMA, and as compared with control group, the expression of PDGF, ERK and α-SMA was increased by 1.4,1.2 and 1.3 fold respectively （P 〈0.01 ,P 〈 0.05 and P 〈 0.01 ）. The increased activity of PKC also promoted the proliferation of HSCs. These effects could be inhibited by the inhibition of PKC activity. Conclusion Changes in the PKC activity could modulate the expression of PDGF and play a role in regulating the proliferation and activation of HSCs.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第7期808-811,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(30400427)
关键词
肝星状细胞
蛋白激酶C
血小板源生长因子
增殖
Hepatic stellate cell
Protein kinase C
Platelet derived growth factor
Proliferation
