| Purpose:To observe the effects of traditional Chinese medicine compound Shenlong Jianji on angiogenesis in IPF rats from the perspectives of multi-level,multi-angle and multi-target based on PI3K/AKT signaling pathway,discuss the mechanism of Shenlong Jianji in intervening angiogenesis in IPF,so as to further explore the action targets of intervention of traditional Chinese medicine compound on IPF.Material and method:1.120 male rats were divided randomly into 4 groups,including Blank Control Group,Model Control Group,Pirfenidone Group and Shenlong Jianji Group,with 30 rats in each;2.The IPF animal model was constructed by intratracheal injection of Bleomycin,and corresponding drugs were given for intervention on the next day of model building.According to the dynamic evolution of IPF pathology,samples were taken at three time points on the 7th、14th and 28 th day after the model was made,and related index tests were carried out;3.Observe the general state of rats;4.Observe the appearance morphology,pathological morphology,and fibrosis morphology changes of rat lung tissues in each group by naked eyes,HE staining and Masson staining;5.Detect and analyze the expression of MVD in the lung tissue of each group of rats by immunohistochemistry and image analysis technology;6.Detect the protein expression levels of PI3 K,p-PI3 K,AKT,p-AKT,HIF-1α and VEGF in the lung tissue of each group of rats by Western Blot experiment method.Results:1.Shenlong Jianji can improve the general status of IPF rats;2.Shenlong Jianji can inhibit the alveolitis in the early stage,the proliferation of collagen fibers in the mid-advanced stages,and reduce fiber deposition so that it can relieve the inflammation of the lung tissues in IPF rats and delay the process of IPF.3.Shenlong Jianji can inhibit pathological angiogenesis by reducing the expression of MVD in the lung tissues of IPF rats.Compared with the Blank Control Group,the expression of MVD in each phase of the Model Control Group was increased.Compared with the Model Control Group,the expression of MVD in each phase of the Treatment Group was decreased in varying degrees.Compared with the Pirfenidone Group,the expression of MVD in each phase of the Shenlong Jianji Group was decreased;4.Shenlong Jianji inhibits angiogenesis by inhibiting PI3K/AKT signaling pathway.Compared with the Blank Control Group,the expression levels of PI3 K and AKT protein in each phase of the Model Control Group did not change significantly,and the expression levels of p-PI3 K and p-AKT protein was increased.Compared with the Model Control Group,the expression levels of p-PI3K、p-AKT protein in each phase of the Treatment Group was decreased in varying degrees.Compared with the Pirfenidone Group,the expression levels of p-PI3K、p-AKT protein in each phase of the Shenlong Jianji Group was decreased;5.Shenlong Jianji can inhibit the expression of HIF-1α and VEGF in the lung tissue of IPF rats,and exert the effect of inhibiting angiogenesis.Compared with the Blank Control Group,the expression levels of HIF-1α、VEGF protein in each phase of the Model Control Group was increased.Compared with the Model Control Group,the expression levels of HIF-1α、VEGF protein in each phase of the Treatment Group was decreased in varying degrees.Compared with the Pirfenidone Group,the expression levels of HIF-1α、VEGF protein in each phase of the Shenlong Jianji Group was decreased.Conclusion:1.Shenlong Jianji can effectively improve the general state of IPF rats,improve the degree of disease,and delay the process of IPF;2.Shenlong Jianji can reduce the expression of MVD in the lung tissue of IPF rats,inhibit pathological angiogenesis;3.Shenlong Jianji can reduce the expression of p-PI3 K and p-AKT to inhibit the PI3K/AKT signaling pathway,thereby reducing the expression of HIF-1α and VEGF in lung tissues to interfere with angiogenesis,thereby delaying the process of IPF. |
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