| Objective:To observe the expression of Wnt/p-catenin Signaling in lung of pulmonary fibrosis in mice, as well as the effect of Thalidomide and Cordyceps sinensis on wnt signaling in this model.Methods:One-hundred and twenty-six healthy male kunming mice were randomly divided into seven groups, including normal control group (N group), model group (M group), thalidomide group (T group), dexamethasone group (D group), high dose of cordyceps sinensis group (H group), middle dose of cordyceps sinensis group (Z group),and low dose of cordyceps sinensis group (L group). Each group has eighteen mice. the mice were respectively intratracheally instilled with bleomycin to pulmonary fibrosis.From the next day, T group was given to mice100mg/kg of thalidomide daily by gastric gavage, D group was given to mice2mg/kg of dexamethasone daily by gastric gavage, H group was given to mice9g/kg of cordyceps sinensis daily by gastric gavage, Z group was given to mice6g/kg of cordyceps sinensis daily by gastric gavage,and L group was given to mice lg/kg of cordyceps sinensis daily by gastric gavage, while N group and M group were given the same volume of normal saline instead until they sacrificed. After treatment each group of mice sacrificed on seventh-day, fourth-day and twenty-eighth day randomly, six mice at a time. To observation the changes of weight in mice, the weight of mice were recorded every three days; take the left upper lung of mice, the level of hydroxyproline was detected with alkaline hydrolysis methods; take the left lower lung of mice, histological changes of lung in mice were evaluated by HE staining; take the right lung of mice, the representative factor WISP-1and MMP-7mRNA of WNT/β-catenin signaling were evaluated by real-time Polymerase Chain Reaction (RT-PCR).Results:1. The changes of weight in mice:Compared with the mice of normal group, the weight growth in mice of pulmonary fibrosis groups become slow, and the model group has the slowest rapid; Compared with the dexamethasone group, the weight growth of thalidomide group was faster; in different dose of cordyceps sinensi groups, the weight growth of high dose of cordyceps sinensis group has increased at the soonest, while the low dose of cordyceps sinensis group has increased lowest, Compared with the dexamethasone group they all increased higher.2. The level of hydroxyproline:①The effect of thalidomide on pulmonary fibrosis lung:at each point of time, the model group has the highest content of hydroxyproline, while the normal group has the lowest. Compared with the dexamethasone group, the level of hydroxyproline in thalidomide group declined, and it has declined significantly on28th day (P<0.05).②The effect of cordyceps sinensis on pulmonary fibrosis lung:at each point of time, the model group has the highest content of hydroxyproline, while the normal group has the lowest. In different dose of cordyceps sinensi groups, the H group has the lowest content of hydroxyproline, while the L group has the highest. The level of hydroxyproline of every group has declined on28th(P<0.05).3. Histological changes of the lung:the pulmonary fibrosis groups have typical pulmonary fibrosis changes. The structural of the lung damaged, large amounts of collagen fiber hyperplasia. After treatment with drugs, the lung has a different degree of change, the part of the lung structural was respaired, collagen hyperplasia was decreased obviously.4. The changes in the expression of WISP-1and MMP-7mRNA:①The effect of thalidomide on pulmonary fibrosis lung:at each point of time, the model group has the highest expression of WISP-1and MMP-7mRNA, while the normal group has the lowest. Compared with the dexamethasone group, the expression of WISP-1和MMP-7mRNA in thalidomide group declined, and it has declined significantly on28th day(P<0.05).②The effect of cordyceps sinensis on pulmonary fibrosis lung:at each point of time, the model group has the highest expression of WISP-1and MMP-7mRNA, while the normal group has the lowest. In different dose of cordyceps sinensi groups, the H group has the lowest expression of WISP-1and MMP-7mRNA, while the L group has the highest. The expression of WISP-1and MMP-7mRNA of every group has declined on28th day (P<0.05).Conclusion:1.The expressions of Wnt/β-catenin signaling increased significantly in model groups, it may be involved in the pulmonary fibrosis.2.The extent of pulmonary fibrosis in T group reduced significantly, it maybe has effects on pulmonary fibrosis by decline the WNT signaling.3. The extent of pulmonary fibrosis in cordyceps sinensis groups, it maybe has effects on pulmonary fibrosis by reducing the WNT signaling, and the high dose of Cordyceps sinensis has more stronger effects than lower dose on pulmonary fibrosis. |
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