| To discuss the effects of individual and combinated administration of N-acetyleysteine(NAC) ,Budesonide and theophylline and the interaction between HAT/HDAC andCOPD, tin rats with chronic obstructive pulmonary disease(COPD) by observing their effecton the oxidation-deoxidization an process and hypoxia inducible nuclear factor kappaB (NF-κ B) ,histone deacetylace(HDAC) in lungs.[methods] 36male Wistar rats were randomly divided into six groups. The normal control group(C), the model group(M), the NAC group(N) ,the NAC combinated with Budesonide group(NB), Budesonide group(B) and Budesonide combinated with theophylline group(BT) . Rats in group M, N ,NB,B and BT were established by intratracheal instillation of lipopolysaccharide twice each day and exoposed to cigarette smoke daily. The group N was treated with NAC by 400mg/kg each day, while rats in group NB reveived both of NAC and Budesonide each day. group B was treated Budesonide each day and group BT was revieved both Budesonide and theophylline. At the end of fourth week, when COPD model was established, items below should be observed:1. Observe the pathomorphological changes in pulmonary with microscope.2. Total and differential cell counts in BALF were done.3. The levels of malonic dialdehyde (MDA) and peroxide dismutase (SOD) and anti-ros in serum were measured with biochemical assays techniques.4. The expression ofNF- κ B and HDAC2 in pulmonary tissue was observed by immunohistochemistry technique.5. observed the expression of HDAC2 in lung through RT-PCR.6. Bronchial morphological changes were observed by scan-electron microscopy. [Results]1. The model group rats shared specific pathological features in trachea, bronchi and lung tissues with that of human chronic bronchitis and obstructive emphysema. Observed underthe light microscope, alveolar tube and alveolar in the HE stained section of emphysema model group were obviously expanded, the walls of alveolar became thin and were broken. The brokenness of alveolar walls and the expand of alveolar cavity in the NAC group, NB group and BT group were more alleviatory than those in the model group. Compared with normal control group and B group, MLI and MAA significantly increase, MAN significantlydecreased (P<0.01 and P<0.01, respectively) in model group; Compared with modelgroup, MLI and MAA significantly decreased (P<0.05 and PO.01, respectively), MAN significantly decreased (P<0.01) in NAC group ,BT group and combinational administration of NAC and Budesonide group.2. Total and differential cell counts in BALF Compared with normal group, total white cell counts and neutrophils in BALF in model group, NAC group and combinational administration of NAC and Budesonide group and BT grouop were significantly increasedthan that in normal group (P < 0.05). Compared with the normal group, the ratio ofmonocyte-macrophage in other five groups decreased obviously, while the monocyte-macrophage count increased. Compared with NAC group, total white cell and neurophils in BALF in NB group an BT group decreased significantly. The ratio ofmonocyte-macrophage in NB group almost returned to normal (P < 0.05).3. The level of SOD ,anti-ROS and MDA in serum Compared with normal group, the level of SOD and anti-ROS in serum of model group ,NAC group and B group decreased significantly while that of MDA increased significantly. There was no difference between the contents of NB group and normal group.4. The expression of NF- k B and HDAC2 in pulmonary tissue Compared with normal group, in which NF- k B was weakly positive in nuclear and negative in cytoplasm, rats in model group and B group expressed NF- k B positive in both of cytoplasm and nuclear. NF- k B in NAC group was positive in nuclear and weakly positive in cytoplasm, while in NB group and BT group it was weakly positive in both of nuclear and cytoplasm. HDAC2 was expressed positive in normal group in pulmonary tunica media, epithelial cells of alveolar and some inflammatory cells,and their positive expressions are exclusively innucleus. Compared with normal group , in model group and B group, HDAC2 was expressed weakly positive only in nucleus. HDAC2 in NAC group was positive in nuclear,and in NB group and BT group,the expression of HDAC2 was positive exclusively in nucleus.5. echogenic band was read by software Alpha Innotech ,and the ratio of each band to 3 -actin was used to be as the expression of HDAC2. other five group were all increased significantly compared to control group.5. Scan Electron Microscope (SEM) image of bronchial morphological change In model group, B group,NAC group and NB group, the epithelial cilia cohered, fell down, partially fell off , accompanied by increasing number of hypertrophic goblet cells and large number of infiltrating inflammatory cells. It was more obvious in model group and B group than in NAC group and NB group. The epithelial cilia of rats in normal group lined regularly, the goblet cells in good situation without any inflammatory cells infiltrating.[ Conclusion ] 1 Smoking can induce strongly expression of and NF- k B, cause the decreaseof HDAC and the inhibitition of glucocorticoid ,cause inflammation in lung and lead to forming of COPD.2 NAC can relieve chronic bronchitis and emphysema, decrease the degree of cigarette smoke inhibiting HDAC .3 Combinated administration of NAC and Budesonide has better effect than individual NAC.4 Low-doses theophylline combined with Budesonide can improve the activity of HDAC2 and inhibit the inflammation in lung. |
