前列腺素E_2和肿瘤坏死因子-α对肺成纤维细胞第二信使的影响

Effect of prostaglandin E_2 and the tumor necrosis factor-α on second messenger of lung fibroblast

目的 探讨前列腺素E2 (PGE2 )和肿瘤坏死因子 α(TNF α)对鼠肺成纤维细胞第二信使的影响,为细胞因子治疗尘肺提供新靶点。方法 在原代培养的小鼠肺成纤维细胞中,加10ng/mlTNF α或10ng/mlTNF α联合不同剂量的PGE2 细胞因子,在不同的观察时点,用放射免疫分析法检测肺成纤维细胞内环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)和1,4 ,5 三磷酸肌醇(IP3 )等第二信使的量变模式对肺成纤维细胞增殖效应的影响。结果 在12 0s时点,10ng/mlTNF α与10 0 0pg/mlPGE2 联合时,成纤维细胞内IP3 的计数值最高[(76 .33±7.10 )CPM];而cAMP/cGMP比值较低时(作用时点2 4h ,4 .2 9) ,对成纤维细胞的增殖效应最强;当10ng/mlTNF α与5 0 0pg/mlPGE2 和2 0 0 0pg/mlPGE2 联合时,成纤维细胞内IP3 的CPM值明显降低:作用12 0s时,CPM值分别为2 7.0 0±3.0 0、6 1.0 0±2 .6 5 ;cAMP/cGMP比值在作用2 4h时(分别为3.5 0、9.83) ,成纤维细胞的增殖效应明显下降。结论 一定剂量的PGE2 能提高肺成纤维细胞内cAMP/cGMP比值,拮抗TNF

Objective To Study the effect of prostaglandin E 2(PGE 2) combined with tumor necrosis factor-α(TNF-α) on the second messenger of mouse lung fibroblast in order to research new target point for cytokine to treatment of pneumoconiosis. Methods The lung fibroblasts of breed mouse were primarily cultured.10 ng/ml TNF-α and 10 ng/ml TNF-α combined with PGE 2 of different doses were added to culture medium of fibroblasts.The γ value and CPM value were respectively measured for second messenger of cAMP,cGMP and IP 3 of fibroblast by immune-radio assay at different observation time. Results When treated with 10 ng/ml TNF-α+1 000 pg/ml PGE 2 at 120 s time points,the CPM value of IP 3 of fibroblast was the maximal value[(76.33±7.10)CPM];when cAMP/cGMP ratio declined to 4.29 at 24 h time point,the effect of fibroblast proliferation was the strongest;when 10 ng/ml TNF-α +500 pg/ml PGE 2,and 2 000 pg/ml PGE 2,the CPM value of IP 3 of fibroblast were 27.00±3.00 and 61.00±2.65 respectively at 120 s time point,cAMP/cGMP value were 3.50 and 9.83 respectively at 24 h,the effect on fibroblast proliferation were obviously lower. Conclusion Certain dose of PGE 2 could raise the ratio of cAMP/cGMP of fibroblast,and antagonize the proliferation effect of TNF-α on fibroblast .