干扰cAMP表达对TNF-α致鼠肺成纤维细胞增殖的影响  

Effects of cAMP Interference on TNF-α Induced Rat Lung Fibroblast Proliferation

作　　者：李慧[1] 李素平[1] 葛伦睿[1] 孟昭伟[1] 

机构地区：[1]山西医科大学公共卫生学院,山西太原030001

出　　处：《工业卫生与职业病》2011年第3期145-148,共4页Industrial Health and Occupational Diseases

摘　　要：目的通过体外实验探讨肿瘤坏死因子-alpha(TNF-α)对鼠肺成纤维细胞增殖的信号转导机制。方法实验分为5组:空白对照组;TNF-α组;TNF-α分别联合10-7、10-6和10-5mol/L佛司克林(FSK)组。SD大鼠乳鼠肺成纤维细胞进行传代培养,刺激培养肺成纤维细胞4和24 h后,用氯氨T法检测细胞培养上清液中羟脯氨酸的含量;用荧光细胞免疫组化技术测定成纤维细胞内磷脂酶C(PLC)和蛋白激酶C(PKC)的表达;用酶联免疫吸附试验(ELISA)检测成纤维细胞内环-磷酸腺苷(cAMP)的含量。结果在同一时间点,单独TNF-α组较空白对照组羟脯氨酸含量和PLC、PKC表达水平升高,TNF-α联合FSK后较单独TNF-α组羟脯氨酸含量和PLC、PKC表达水平降低,差异有统计学意义(P<0.05);PLC、PKC结果还显示,随TNF-α联合FSK作用时间的延长,PLC、PKC表达减弱,差异有统计学意义(P<0.05)。TNF-α诱导PLC和PKC表达的同时,也能提高cAMP水平,与空白对照组比较,差异有统计学意义(P<0.05)。当TNF-α联合FSK后能进一步促进cAMP表达,与单独TNF-α组比较,差异有统计学意义(P<0.05)。结论 TNF-α对成纤维细胞增殖有促进作用。FSK能以剂量依赖关系提高cAMP水平,降低TNF-α诱导的成纤维细胞增殖信号PLC和PKC的表达,从而使羟脯氨酸含量降低。Objective To discuss effects of tumor necrosis factor（TNF-α）on signal transduction mechanism of rat lung fibroblasts proliferation by experiments in vitro.Methods 5 groups were experimented i.e.blank control group,TNF-α group,TNF-α joint 10-7,10-6,and 10-5mol/L FSK groups.SD neonatal rat lung fibroblasts were sub-cultured.After stimulating lung fibroblasts for 4 h and 24 h,hydroxyproline content in the supernatant of cell culture was detected with chloramine T method.PLC and PKC expressions were determined by fluorescent cell immuno-histochemical technique.cAMP content in fibroblasts was detected by ELISA test.Results At the same time point the hydroxyproline contents and the expressions of PLC and PKC in the group of TNF-α alone were elevated,however,after TNF-α combined with FSK,the hydroxyproline contents and the expressions of PLC and PKC became lower than those of the group of TNF-α alone,the difference was significant（P0.05）.The results of PLC and PKC expressions also showed that as the reaction time of combination of TNF-α with FSK prolonged the expressions of PLC and PKC were weakened,the difference was significant（P0.05）.At the time of TNF-α inducing the expressions of PLC and PKC,the level of cAMP was elevated simultaneously;the difference was significant（P0.05）.When TNF-α combined with FSK,the expression of cAMP would further be stimulated in comparison with that of the group of TNF-α alone（P0.05）.Conclusions TNF-α could promote the proliferation of fibroblasts.FSK could increase the level of cAMP in dose-dependent relationship.FSK could also reduce the TNF-α induced signals of fibroblast proliferation（expressions of PLC and PKC）,so as to lower the content of hydroxyproline.

关 键 词：肿瘤坏死因子-α 环-磷酸腺苷 磷脂酶C 蛋白激酶C 佛司可林 

分 类 号：R135.2[医药卫生—劳动卫生]