The Mechanism And Clinical Analysis Of The Effect Of Icariin On Pulmonary Fibrosis Through Wnt/β-catenin Signal Pathway

Objective: Pulmonary fibrosis(PF)is a pathological change form in the end stage of interstitial lung disease,which has fast progress and high mortality,but its pathogenesis is unknown and the clinical treatment effect is not good.In recent years,more and more experiments have proved that oxidative stress plays an important role in the development of pulmonary fibrosis,and the imbalance of cell oxidation and antioxidation is the key to the formation and progress of pulmonary fibrosis.Related studies have shown that Wnt/β-catenin signal pathway is abnormally activated in the early stage of pulmonary fibrosis,the imbalance of oxidative stress,activation and proliferation of lung fibroblasts,aggravating the progress of pulmonary fibrosis.In order to further understand the mechanism of the Chinese tonifying medicine Epimedium on pulmonary fibrosis,the experiment selected icariin to act on human embryo lung fibroblast MRC-5.By observing the changes of the related gene proteins in the Wnt/ β-catenin signal pathway to explore the anti-pulmonary fibrosis mechanism of Epimedium and provide experimental basis for clinical medicine.Methods: According to the experimental requirements,human embryonic lung fibroblast MRC-5 is the object of study,icariin is an intervention drug,and the experiment was divided into 4 groups: 1.blank group(MRC-5)2.control group(MRC-5+ Wnt1.)3.treatment group(MRC-5+ Wnt1+ icariin)4.The pre-protective group(MRC-5+ Icariin +Wnt1).Based on the successful cultivation of human embryonic lung fibroblasts,using the method of CKK-8 to measure the maximum non-toxic concentration of icariin,the method of Cell biochemical detection to detect the activity of SOD,CAT,the oxidation index change of MDA and GSH-PX of the 4 groups,the method of Western Blotting and QPCR to detect the expression of core protein β-catenin and the substream protein TCF/LEF,cyclin Dl,fibronectin and MMP-7 in the Wnt/ β-catenin Signal pathway,and the method of Immunofluorescence to detect the expression of collagen I and collagen III.Results: After the intervention of icariin in MRC-5,it can significantly increase the concentration of SOD,CAT GSH-PX,decrease the concentration of MDA,effectively enhance the antioxidant level of cells,and he effect of icariin preprotection is better than the treatment of drug use.While icariin intervention can significantly reduce the aggregation of β-catenin in the nucleus,through the Real time PCR method and Western Blot method,it was found that the treatment of drugs could not reduce the expression of TCF,and the rest of the experiments confirmed that the expression of downstream genes TCF,LEF,cyclin Dl,fibronectin and MMP-7 showed a downward trend,and effectively inhibited the proliferation of collagen I and collagen III at the same time.It indicates that icariin can effectively inhibit the activity of Wnt/ β-catenin signal pathway,inhibit fibroblast activation and collagen proliferation after intervention,and the effect of icariin pre-protection is better than the treatment of drug use.Conclusion: Icariin can improve the antioxidant capacity of MRC-5 in human embryonic lung fibroblasts.By inhibiting the activity of Wnt/ β-catenin signal pathway,it can effectively inhibit the activation of MRC-5,reduce the transformation of lung fibroblasts to myofibroblasts,and alleviate the progression of pulmonary fibrosis.Through experimental comparison,the pre-protective drug is superior to the treatment of drugs,which provides a solid theoretical basis for the prevention and treatment of pulmonary fiber in the clinical Epimedium.