| ObjectivesTo summarize the experience of the clinical treatment on children with pulmonary fibrosis by the Modified Xiaokuxin Soup, and to observe the effect of treatment on mice with BLM-induecd pulmonary fibrosis by the Modified Xiaokuxin Soup.To investigate the pathogenesis of pulmonary fibrosis and mechanism of the treatment on pulmonary fibrosis in children by the Modified Xiaokuxin Soup.Methods72C57BL/6female mice were randomly divided into control group, model group, prednisone group and high,moderate and low Modified Xiaokuxin Soup groups, n=12. Groups of model group, Chinese medicine group and western medicine group with bleomycin(40mg/kg)200μl intraperitoneal injection5times were modeled into pulmonary fibrosis in mice, and the control group with normal saline instead. Mice in each group were gavaged from the day after molding, the three doses groups of traditional Chinese medicine were respectively given the Modified Xiaokuxin Soup granules,4.6g/kg,2.3g/kg,1.15g/kg; the western medicine group were given prednisone5mg/kg; the model group and control group were given to0.5%CMC10ml/kg once a day. On the14th and28th day after the consecutive gavage,mice were killed in batches. Then take full wet lung weight, and take the right lung tissues were placed in4%formaldehyde solution for fixation, conventional dehydrat ion, embedded in paraffin to be consecutive tissue sections, whicd were then conducted Suwood alcohol-eosin (HE staining) trichrome stain(Masson stain) and immunohistochemical staining. Morphological changes of the biopsy lung tissue and expression of TGF-β1, ICAM-1, NF-κβ in each group were observed. And the integrated optical densit(IOD) and mean optical density (MOD) value of TGF-β1, ICAM-1, NF-κβ in lung tissue of mice in each group were compared by image analysis.Results1. The impact of the lung mass coefficient:Lung mass coefficient in the model group on14th and28th days were significantly higher than the control group after treatment, there was a significant statistical significance (P<0.01); the western medicine group compared with the control group were statistically s ignifican (P<0.05); Chinese medicine high dose group on14th and28th days were lower than the model group,which was statistically significan(P<0.05); there is no significant differencebetween the low-dose of Chinese medicine group and the control group on28th day(P>0.05),and the Chinese medicine high dose group was lower than western medicine group, the difference was statistically significant (P<0.05).2. The impact on Lung tissue morphology:①HE staining:in the control group, there was not changes in pulmonary alveolitis and fibrosis,and the alveolar structure was integrity, the leakage in alveolar cavity was not significant. In the model group,on the14th day, inflammatory cell gathered together around the bronchioles and perivascular, the alveolar septa was widened,leakage of the alveolar cavity was significant; while on the28th day, there are large areas of consolidation fibrosis area,part of the alveolar wall destruction, collapse of the alveolar space,alveolar wall thickening, alveolar septal fibrosis significant.②Masson staining:On the14th day, in the model group, alveolitis was significant and fibrosis was severe. The alveolar space was filled with exudate and the shedding of a variety of inflammatory cells, there was visible apparent fiber hyperplasia lesions in the alveolar septa. On the28th day, around the alveolar wall, interval bronchus and small blood vessels, a large number of fibers were proliferated. In summary, in three of the Chinese medicine and Western medicine group, the alveolitis and fibrosis degree reduced compared with the model group, among that Chinese medicine high dose group was particularly evident.3. The impact on TGF-β1expression:on the14th and28th days, in the model group and control group, IOD and MOD of the cells were significantly increased (P<0.01); IOD and MOD of the cells of the three dose of Chinese medicine group was significantly lower compared with the model group (P<0.01). Chinese medicine high dose group compared with the western medicine group,on the14th day,the difference of IOD was not significant (P>0.05),while on the28th day was lower than western medicine group (P<0.05). There was no difference between the Chinese medicine high dose group and the western medicine group in MOD(P>0.05). Chinese medicine high dose group, western medicine group and the control group, the difference in IOD and MOD of the cells was not significant (P>0.05).4. The impact on ICAM-1expression:The IOD of the cells in the model group was significantly increased than the control group (P<0.01); while the three dose of the Chinese medicine groups were significantly reduced compared with the model group in IOD and MOD of the cells (P<0.01); there was significant difference between the high dose of Chinese medicine group and the western medicine group on the14"’and28th days (P<0.01),which was the same as the MOD between the low dose of Chinese medicine group and the western medicine group on the28th day. The differences were significant in each group compared with the control group in MOD on the14th day (P<0.01), while there was no difference between the high dose of Chinese medicine group and the control group on the28th day (P>0.05); the three dose of the Chinese medicine groups were significantly reduced compared with the model group (P<0.01);and the difference was significant in the high dose of Chinese medicine group compared with the western medicine group (P<0.01).5. The impact on NF-κB expression:The IOD and MOD of the cells in each group increased significantly than the control group on the14th and28th days (P<0.01). Compared to the model group, IOD and MOD of the three dose of Chinese medicine group reduced significantly compared with the model group in the IOD and MOD of the cells (P<0.01). IOD in each dose of Chinese medicine group was decreased significantly than the western medicine group on the14th day (P<0.01), and in the high dose of Chinese medicine group IOD and MOD was decreased significantly (P<0.01), with the middle dose of Chinese medicine group most obvious (P<0.01).Conclusions:1. We at tribute pulmonary f ibrosis in chi ldren to " lung Bi" category in Chinese Medicine. The actual situation of the disease is a mixture of complex pathogenesis, with a total deficiency oriented to lung, blood stasis and phlegm blockage of the lung as a standard network. The treatment under the principle that "non-Xin Yu is not open, non-bitter the fire did not fall", uses acrid bitter drop and adjusting qi of the law, to clear the lungs phlegm, cough and asthma and live stas is meridians, with the modified Xiaokuxin Soup added and subtracted, giving dialectical treatment of children with pulmonary fibrosis, achieving a certain effect.2. In the way of intraperi toneal injection of bleomycin (40mg/kg),200μ1,5times, we had successfully copied pulmonary fibrosis in a mouse model, whose pathological changes was in line with the development of the early alveolitis and advanced pulmonary fibrosis.3. The modified Xiaokuxin Soup could improve the pulmonary fibrosis in mice, and can reduce lung mass coefficient.It significantly reduced the alveolitis and pulmonary fibrosis.4. The expression of TGF-β1, ICAM-1,NF-κβ in pulmonary fibrosis lung tissue of mice induced by Bleomycin increase significantly, showed that TGF-β1, ICAM-1and NF-κβ involved in the development of pulmonary fibrosis process, and in which may be an important link.5. The modified Xiaokuxin Soup may be down lung tissue of TGF-β1, ICAM-1, NF-κβ expression and play the role of anti-fibrosis. |
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