The Effect Of TAL On The Expression Of TNF-α In Alveolar Macrophage Of Chronic Bronchitis Rats And Parts Of Mechanism

Chronic Bronchitis (CB) is a common respiratory system disease,because of infection or non-infection factors. Its pathological features are bronchial gland hyperplasia, increased mucus secretion. Clinical manifestations were more than two years in a row, each sustained for more than three months of cough, sputum, or asthma symptoms. Disease progression can be complicated by obstructive pulmonary emphysema, pulmonary heart disease, serious impact on the labor and health .At present, people pay more attention to anti-inflammatory, relieving cough on curing the disease, however, it is hard to find an effective medicine. While the conditions of immune function play a more important role in the process of CB. Therefore, to regulate the disorder of neuroendocrine-immuno-function may the key method to prevent and cure the Chronic Bronchitis.The leaf of Eriobotrya japonica (Thunb.)Lindl has a long history of medicinal use in China in a variety of inflammatory conditions especially CB therapy. According to our previous study, the triterpene acids extracted from Eriobotrya japonica (Thunb.)Lindl Leaf was the effective component and had therapy effect on chronic bronchitis. In addition, the characters of this medicine are the high performance and low toxicity.Alveolar macrophage is widely distributed in alveolar and airway epithelial surface, is the localized main immune cells of lower respiratory tract. It not only has the phagocytic function, but also secrete a variety of active substances, such as interleukin (IL), tumor necrosis factor-α(TNF-α), nitric oxide (NO), prostaglandin (PG) and other substances, plays an important regulatory role in physiological and pathological of lungs. According to our previous study, AM has been activated in the LPS combined BCG induced model of chronic bronchitis, and plays an important role. Role of AM is closely related to cytokines and inflammatory mediators. TNF-αis one of the most important immunoregulatory factor. So study the impact and mechanism of TAL in TNF-αof chronic bronchitis model's AM is important. As widely known that MAPK play a key role in inflammatory reaction, we use the AM and special inhibitors of MAPK signal transduction pathway (ERK,p38 or JNK),as the researching platform and target respectively, to study the MAPK signal transduction pathway in AM, and the factorial design is used to discuss the effect of TAL. It can help us to have a deep view in the molecular mechanism of the curing effect of TAL, meanwhile, we can get the basement for the research of the new medicine. The main content is divided into three sections as follows:1. The relationship between TNF-αand MAPK signal transduction pathway in alveolar macrophage of chronic bronchitis ratsEstablishing the model of CB rats by using BCG plus LPS, We incubated the depurated AM with the special inhibitors of MAPK signal transduction pathway(PD98059,SB203580,Curcumin),to observed TNF-αlevel in AM culture supernatant and the expression of TNF-αby radioimmunoassay and RT-PCR, Western Blot. We observed impact of the three MAPK pathways (ERK, P38, JNK) on the TNF-αexpression. The results suggest that TAL reducing the expression of TNF-αmay be concerned with the inhibition of ERK and p38 signal transduction pathway. It may be one of the mechanism by which TAL has the preventive and therapeutic effects on chronic bronchitis.2. The mechanism of TAL treating chronic bronchitisThe experiement used TAL combined the special inhibitors of ERK,P38(PD98059/SB203580), and the factorial design is used to discuss the effect of TAL. The results of RT-PCR and Western Blot prompted the impact of TAL and PD98059/SB203580 on TNF-αin AM of chronic bronchitis may be synergism. The results of factorial design experiement showed, compared with model group, the TNF-αlevel decreased significantly after TAL or PD98059/SB203580 used alone. Compared with TAL or PD98059/SB203580 used alone group, the TNF-αlevel decreased significantly after TAL combined PD98059/SB203580. Factorial analysis shows there are statistical significance between TAL , PD98059/SB203580 and Interaction of their.This confirmed the impact of TAL and PD98059/SB203580 on TNF-αin AM of chronic bronchitis are synergism. TAL reducing the expression of TNF-αmay be concerned with the inhibition of ERK and p38 signal transduction pathway. It may be one of the mechanism by which TAL has the preventive and therapeutic effects on chronic bronchitis.3. Application of factorial design in study of the curing effects of TAL on chronic bronchitisTwo factorial designs were applied to observe the effects of TAL at different concentrations with PD98059/SB203580 at different concentrations, on the inhibition TNF-αin AM of chronic bronchitis, and the interaction among the each factor. The results of factorial design experiements showed, when the concentration of PD98059 was 150mol/L and the concentration of TAL was 250mol/L, the inhibition effects of TNF-αreached the peak,or when the concentration of SB203580 was 10mg/L and the concentration of TAL was 250mol/L. It may provide experimental evidence for the curing effects of TAL combined the special inhibitors of ERK , P38(PD98059/SB203580)on chronic bronchitis...