| Objective:Pulmonary fibrosis is the final stage of a variety of lung diseases.Traditional medicine moxibustion treatment of pulmonary fibrosis have an established therapy effect,but still lack of in-depth mechanism study.Based on our previous study,this work is to observe the process of pulmonary fibrosis,focused on Wnt/beta-catenin signaling pathway,through the approach of moxibusting the Feishu and Gaohuangshu points against the pulmonary fibrosis process.Methods: 100 of SD male rats,two months and 180 ± 20 g weight,were recruited and divided into four groups: normal control group(N),model group(M),moxibustion group(A)and prednisone group(P).Pulmonary fibrosis rats were made by the injection of bleomycin into the trachea,5 mg/kg once.Since the second day after bleomycin injection,rats of the moxibustion group were moxibusted at Feishu and Gaohuangshu points,once a day and last for 35 days.Rats of the prednisone group were stomach-injection of prednisone 5 mg/kg every day.The model group is fixed the same as the moxibustion group without any treatment.The general index such as breathing,body weight and mental state were recorded every day for each animal.At 7d,11 d,23d and 35 d post-modeling,the rats were randomly selected and sacrificed to dynamically observe the expression of Wnt/ beta-catenin signal pathway in the process of pulmonary fibrosis.HE staining and Masson staining were used to observe the pathological features of lung tissue.The expression of beta-chain protein(beta-catenin)in lung tissue of each group was detected by immunohistochemistry.Enzyme linked immunosorbent assay(ELISA)was used for the detection of beta-catenin,Wnt induced secreted protein-1(WISP-1),metal matrix protein-7(MMP-7),Wnt-3a and other proteins in lung tissue.The dynamic expression of Wnt-3a m RNA levels in each group were detected by real-time fluorescence quantitative PCR.The expression of WISP-1 and MMP-7 in lung tissues of each group was detected by Western blotting.Results: The general conditions of moxibusted rats were obviously improved.Compared with the normal group,the rats of model group were generally in poor condition.Compared with the model group at the same time point,the respiration and body weight of the rats in the moxibustion group were significantly improved.Pathological observation showed that the inflammatory cell infiltration and fibrosis in lung tissue of the rats were significantly reduced.Results of HE staining showed that inflammatory cell infiltration in the interstitial tissue of the model group was significantly increased,and the interstitial tissue of the lung was widened.Compared with the model group at the same time point,the degree of inflammatory cell infiltration in the lung tissue was significantly low(P<0.05).Under the Masson staining,the collagen fibers in the model group were significantly increased,and the degree of fiber proliferation was significantly decreased in the moxibustion group and the same time point model group(P<0.05).In the model group,the expression levels of key factors in Wnt/ beta-catenin signaling pathway were significantly increased.The expression of beta-catenin in lung tissue of model group was significantly increased,and peaked at 23 d and then decreased,compared with the control group(P<0.05).In the model group,the expression of Wnt3 a in lung tissue increased at each time point,and reached the peak after 11 d,and there were significant differences between the two groups(P<0.01).The expression of MMP-7 in model group was higher than those in normal group(P<0.01).The expression of WISP-1 in the model group was higher than that in the normal group,and there were statistical differences at 11 d,23 d and 35 d(P<0.05).Moxibustion treatment can significantly reduce the expression of key factors of Wnt/ beta-catenin signaling pathway.Resulsts of immunohistochemical analysis displayed that the concentration of beta-catenin in rats of M11 were markedly reduced,compared with statistical significance(P<0.05).However,moxibustion group at 23 d compared with the model group was statistically significant(P<0.01),was not statistically difference at 35 d;to detect the expression of beta-catenin m RNA,moxibustion group at 23 d compared with model group,with significant difference(P<0.01),moxibustion group at 35 d compared with the model group,with statistical difference(P<0.05).There was no significant difference between the moxibustion group and the prednisone group at the same time point(P>0.05).The expression of Wnt3 a in the moxibustion group was lower than that in the model group,which was statistically significant at 11 d(P<0.01),and was not statistically significant at day 23 and day 35.There was no significant difference between the moxibustion group and the prednisone group at the same time point(P>0.05).The expression of MMP-7 and WISP-1 in the moxibustion group was significantly lower than that in the model group(P<0.05).There was no significant difference between the moxibustion group and the prednisone group at the same time point(P>0.05).Conclusion: Moxibustion treatment displayed significant anti-fibrosis effects through system regulation of the key factors of Wnt/ beta-catenin signaling pathway. |
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