| Objectives:1.Summarized and analysed the clinical data of children with bronchiolitis obliterans(BO),so as to discussed the distribution law of syndrome elements.2.Observed the changes of lung coefficient,matrix metalloproteinase-9(MMP-9)and matrix metalloproteinase inhibitor-1(TIMP-1)expression in lung tissue of BO rats at different time points,analysed the correlation of lung coefficient with MMP-9 and TIMP-1,to investigate the role of MMP-9 and TIMP-1 in fibrosis.3.Established the rat model of BO with lung deficiency cold syndrome,observed the changes of body weight,anal temperature,toe temperature,autonomic activity,lung coefficient,lung tissue MMP-9 and TIMP-1 expression at different time points,and observed the intervention effect of warming lung mixture on the above indexes,to investigate the therapeutic mechanism of warming lung mixture on BO with lung deficiency cold syndrome.Methods:1.Collected the basic information and data of TCM from children with BO by the method of epidemiological investigation and syndrome element from clinic and ward in Shandong Provincial Traditional Chinese Medical Hospital and Shandong Provincial Hospital from February 2014 to February 2017.Using SPSS22.0 statistical software to establish syndrome four diagnostic information database,to organize and analyze the information.2.Wistar rats were randomly divided into normal group and model group.BO rats model was set up in the model group by diacetyl(DA)airway instillation.The normal group did not do any treatment.The changes of body weight,lung pathology and lung coefficient of rats in two groups were observed at 7,14,21 and 28 days.The expressions of MMP-9 and TIMP-1 in lung tissues were detected by Western Blot.Made the correlation analysis between lung coefficient and MMP-9 and TIMP-1.3.Wistar rats were randomly divided into normal group and model preparation group.The normal group did not do any treatment.Model preparation group was prepared by BO combined fumigation and cold stimulation.After successful modeling,rats in model preparation group were randomly divided into model group,low dose group of Chinese medicine,middle dose group of Chinese medicine,high dose group of Chinese medicine and western medicine group.Begin gavage from the Seventh day after successful mold was made.Normal group and model group were treated with physiological saline 2ml · D-1.Low dose group of Chinese medicine were treated with warming lung mixture by 3.145mg ·kg-1 ·d-1.Middle dose group of Chinese medicine were treated with warming lung mixture by 6.29mg · kg-1 · d-1.High dose group of Chinese medicine were treated with warming lung mixture by 12.58mg · kg-1 · d-1.Western medicine group received dexamethasone 0.08mg ·kg-1 ·d-1+montelukast 1.07mg· kg-1 · d-1.The normal and model groups were dissected at day7,day14,day21 and day28.Each drug intervention group was dissected at dayl4,day21 and day28.The body weight,anal temperature,toe temperature,autonomic activity,pathological changes,lung coefficient,inflammation and fibrosis damage were observed in each group.The expressions of MMP-9 and TIMP-1 in lung tissue were detected by Western Blot.Results:1.(1)BO TCM Syndrome Distribution:The score of disease index was highest in lung,followed by the spleen and heart,kidney,etc.(2)The distribution of TCM syndrome elements in BO:The syndrome of phlegm syndrome was the highest in phlegm syndrome,the second was deficiency of Yang,cold,Qi deficiency,wind,blood stasis,heat and Yin deficiency.(3)The distribution of different integration grades in cold related with the course and trend of disease.The distribution of different integration grades in deficiency of Yang related with the course and trend of disease so as to age and sexist.2.Lung coefficient of rats in model group(13.72 ±0.50,12.64±0.65,11.87 ±0.70,10.52 ±0.67)was significantly higher than the normal group(7.68 ± 0.18,7.23 ± 0.06,7±0.08,6.92 ±0.09),the difference was statistically significant(P<0.01).The degree of inflammation and fibrosis in model group was significantly higher than that in normal group.MMP-9 in lung tissue of rats in the model group(0.695 ±0.007,0.595 ± 0.078,0.615±0.035,0.540±0.071)was significantly higher than the normal group(0.065±0.018,0.068 ±0.012,0.070 ±0.005,0.064 ±0.007),the difference was statistically significant(P<0.01).The lung tissue of rats in TIMP-1 model group(0.655 ± 0.005,0.858 ±0.034,0.783±0.047,0.602±0.051)was significantly higher than the normal group(0.069±0.016,0.071 ±0.006,0.069±0.017,0.068±0.009),the difference was statistically significant(P<0.01).The lung coefficient is related to MMP-9 and TIMP-1.3.The improvement of general situation in middle dose group of Chinese medicine and high dose group of Chinese medicine was superior to model group and Western medicine group,the difference was statistically significant(P<0.05).Lung coefficient in middle dose group of Chinese medicine and high dose group of Chinese medicine was significantly lower than model group,the difference was statistically significant(P<0.01).The level of MMP-9 and TIMP-1 in middle dose group of Chinese medicine,high dose group of Chinese medicine and Western medicine group was significantly lower than model group,the difference was statistically significant(P<0.01).Conclusion:1.The main lesion site in BO is lung,closely related with spleen.The main pathological characteristics are cold,Yang deficiency and Qi deficiency.The main pathological products are sputum and blood stasis.It is suggested that the main syndrome type of children with BO are lung deficiency cold syndrome,Yang deficiency syndrome and asthenia syndrome of lung and spleen.Sputum and blood stasis are important pathogenic factors.2.The content of MMP-9 and TIMP-1 in lung tissue of BO model rats increased significantly,and related with lung coefficient,suggested that MMP-9 and TIMP-1 participate in the pathogenesis of BO,and may play an important role in BO airway inflammation and fibrosis.3.The middle and high dose of warming lung mixture can significantly improve the general situation of rats with BO in lung deficiency syndrome,and can reduce lung coefficient.The mechanism of action may related to the reduce of the expression of MMP-9 and TIMP-1. |
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