| Objective:this experiment through the observation of Dan Bei Yi Fei Fang on human embryo lung fibroblast proliferation were inhibited,observation and statistics of different concentrations of Dan Bei Yi Fei Fang solution on the effect of human embryonic lung fibroblast cells collagen,to study whether human embryonic fibroblast cells and inhibit the role of intervention,to prove the beneficial effect of Tempe lung on pulmonary fibrosis and provide experimental basis for Dan Bei Yi Fei Fang decoction in the treatment of pulmonary fibrosis.Method:Human embryonic lung fibroblast cell 1.in vitro,with different concentrations of Dan Bei Yi Fei Fang for treatment,divided into normal control group,the Dan Bei Yi Fei Fang1×10-3 mg/ml group,the Dan Bei Yi Fei Fang 1×10-2 mg/ml group,the Dan Bei Yi Fei Fang1×10-1 mg/ml group,Dan Bei Yi Fei Fang 1 mg/ml,at the temperature of 37?,concentration of incubator 5%,culture time were 24 h,48 h and 72 h.The microscope observation and distinct morphological changes were recorded,and then the application of MTT method,recording and detection of different concentrations of Dan Bei Yi Fei Fang in different time,and lung fibroblasts in a dual wavelength?490 nm,630 nm?the value of A.By recording and comparing the value of A time and dose effect chart,finally compare the effects of different concentration and different time Dan Bei Yi Fei Fang on lung fibroblast proliferation.2.by MRC-5 cells cultured in vitro,and the cultured cells were divided into four groups:normal control group,the dexamethasone group(5×10-2 mg/ml),the Dan Bei Yi Fei Fang in low concentration group(1×10-3 mg/ml),the Dan Bei Yi Fei Fang in high concentration group(1×10-1 mg/ml).By cell smear immunohistochemistry method to observe the intracellular collagen type I?COL I?,collagen type III?COL III?expression,at the same time using Image Pro-Plus image analysis system to calculate the corresponding groups of COL I and COL III IOD expression.3.by MRC-5 cells cultured in vitro,and the cultured cells were divided into four groups:normal control group,the dexamethasone group(5×10-2mg/ml),the Dan Bei Yi Fei Fang in low concentration group(1×10-3mg/ml),the Dan Bei Yi Fei Fang in high concentration group(1×10-1 mg/ml),while using real-time the amount of PCR,for each group of human embryonic lung expression of collagen m RNA fiber cells were measured.Result:1.Dan Bei Yi Fei Fang groups:1×10-3 mg/ml group,the mg/ml group,1×10-2 and 1×10-1mg/ml group,the 1 group mg/ml,were incubated respectively at different time by hierarchical and MRC-5 cells:24h,48h,72h,the results show that with the the concentration of Dan Bei Yi Fei Fang in increasing,MRC-5 were incubated with the continuous extension of time,found that the cell proliferation inhibition rate changed obviously.At the same time,Dan Bei Yi Fei Fang in 1mg/ml group cell proliferation inhibition rate was the highest;the same concentration,Dan Bei Yi Fei Fang in time was 72h,the proliferation inhibition rate was also the highest value;so it can be concluded that the inhibition rate of proliferation of Dan Bei Yi Fei Fang 1 group mg/ml cells gradually increased with time gradually change.2.was used on the expression of COL 1 protein and COL III protein changes in MRC-5cells were analyzed by Image method,Pro-Plus image analysis system to calculate each COL 1 protein and COL protein expression of IOD III,through statistical analysis we found that dexamethasone group,Dan Bei Yi Fei Fang in low concentration group(1×10-3 mg/ml),Dan Bei Yi Fei Fang in high concentration group(1×10-1 mg/ml)were MRC-5 cells COL 1protein and COL III protein expression of IOD compared with the normal control group IOD decreased,the difference was statistically significant?P<0.05?;Dan Bei Yi Fei Fang in high concentration group(1×10-1 mg/ml)compared with dexamethasone group,no significant difference?P>0.05?.3.by real-time quantitative PCR method,the relative expression of MRC-5 cells in COL I m RNA,COL III m RNA was measured,results showed that dexamethasone group,Dan Bei Yi Fei Fang in high concentration group,Dan Bei Yi Fei Fang in low concentration group cells in MRC-5 m RNA COL I and COL III m RNA expression and normal control group.The relative expression levels were reduced in different degrees,and the difference was statistically significant?P<0.05?,Dan Bei Yi Fei Fang in high concentration group was statistically significant with Dan Bei Yi Fei Fang in low concentration group and dexamethasone group were compared?P<0.05?.Conclusion:1.Dan Bei Yi Fei Fang by inhibiting the proliferation of human embryonic lung fibroblast MRC-5 cells,the decrease in the lung deposition,contributes to the prevention and treatment of pulmonary fibrosis.2.Dan Bei Yi Fei Fang with anti pulmonary fibrosis,the mechanism may be through inhibiting the expression of MRC-5 cell COL I m RNA,COL III m RNA,and inhibit the synthesis and secretion of collagen cells,reduce collagen deposited in the lungs,achieve the purpose of prevention and treatment of pulmonary fibrosis. |
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