| Object: Interstitial pulmonary fibrosis is a kind of disease mainly characterized by inflammation and structural destroy of alveolus, which finally changed into interstitial fibrosis. The therapeutic approach to this not particularly effective because of its unclear mechanism. Tranditional Chinese medicine has reached a several achievements in improving symptoms and reducing death rate, and it's necessary to summarize and improve the methods of TCM for this disease. Huaxian decoction(HD), a prescription of tonifying qi and regulating yang, smoothing qi activing blood and eliminating sputum, has shown a bright future in clinical study of pulmonary fibrosis. In this article, the rat model was induced by blyomicin and we evaluated HD's curative effect objectively and explore its way and mechanism by observing the changing of apoptosis of ATⅡcells, the content of TNF-αand IL-6 in sera, the expression of CTGF, the level of SOD, GSH-PX and MDA in lung, the morphological alternation of the lung tissue.Methods: Two hundreds and forty healthy Wistar rats, weight from 180 to 220gram were randomly divided into six groups: normal control group(group A, n=48), model group(group B, n=48), prednisone group(group C, n=48), Chinese drugs of high(group D, n=48), moderate (group E, n=24) and low dosage (group F, n=24). The rat model was induced by blyomicin and respective medicine was given to each group. Every group was subdivided into the three groups which was killed in the seventh, fourteenth, and twenty-eighth day respectively.①The rat's lung tissue section was observed under light microscope and electron microscope to study the morphological alternation.②Find the changes of apoptosis of ATⅡcells by flow cytometry.③Assay the sera level of TNF-αand IL-6 by ELISA method.④Detect the expression of CTGF in the lung tissue with immunohistochemical method of analysis.⑤TBA method and colorimetric method were used to measure the content of SOD, GSH-PX and MDA in lung tissue respectively.Results:①Changes of pathologic morphosis under light microscope in the rat lung fibrosis model: in the 7th day after model was made, interval between alveolus became thicker,, the number of fibroblast increased and alveoli in the focus of infection were atrophied and lots of inflammation cells infiltrated; in the 14th day after model was made, interval between alveolus became thicker and thicker, fibroblast and collagen matrix increased markedly; in the 28th day after model was made, alveoli structure was damaged, the wall of alveoli became thicker remarkably, interstitial fibrotic cicatrix came into being and capillary blocked. Changes under electron microscope in the 7th and 14th day showed that mitochondria of typeⅡalveolar (epithelial) cells became hyperplastic and swollen and its cristae decurtated, collapsed and vanished. Quantities of lamellar body decreased and became vacuole. Microvilli damaged, lessened and disappeared. In the 28th day, quantity of typeⅡalveolar (epithelial) cells decreased more than that in the 7th day or in the 14th day. Cristae in mitochondria and lamellar body damaged and degenerated like vacuole. In the 7th day, amelioration of pathological changes from the most obviously to the most slightly was D, C, E, F, and that in the 14th day was D, E, C, F, that in the 28th day was D, E, F, C. There was significant difference among Szapiel grade of alveolitis and fibrosis in each group (P＜0.01 or 0.05).②the changes of apoptosis of ATⅡcells in group C, D in each phase was lower than that in groupB (P＜0.05).③The sera level of TNF-αand IL-6 in group C, D, E, F in each phase was lower than that in group B respectively(P＜0.01) in the 7th, but only that in group D, E was higher than that in group B in the 14th -28th (P＜0.01 or 0.05).④The expression of CTGF in the lung tissue in the group C, D, E in each phase was lower than that in groupB respectively (P＜0.01 or 0.05).⑤Content of MDA in lung tissue in group C, D, E, F in each phase was lower than that in group B respectively(P＜0.01or0.05), while that of GSH-PX and SOD was higher(P＜0.01, 0.05). In the 7th and 14th day, content of MDA in lung tissue of degressive sequence was F, C, E, D, and in the 28th day was C, F, E, D (P＜0.01or 0.05), while that of GSH-PX was opposite.Conclusion:①The model was made successfully according to its corresponding manifestation of pathology to human's lung fibrosis and other reports.②HD has therapeutic effect on lung fibrosis in each phase.③HD can decrease the apoptosis of ATⅡcells.④HD can regulate the disbalance of cytokine such as decreacing sera TNF-α, IL-6 level.⑤HD can inhibit the expression of CTGF in lung tissue.⑥HD can decrease harmful free radical and increase beneficial free radical scavenger in lung tissue. |
PDF Full Text Request