From Phlegm Blocking Lung Rat Model Of The Sputum Evil Pathogenic Mechanism

Object: This paper demonstrates the essence of phlegm, which causes disease by observing the change of membrance fluidity and ultrastructural changes, the secretion of nitrogen oxide(NO)and tumornecrosis factor- a (TNF-α ) of alveolar macrophage(AM),the expression of CD11a,CD11b,CD18,CD54, CD62L of AM and white cell in blood; the expression of nuclear factor-kappaB (NF-κB) and cyclooxygenase-2(COX-2)mRNA in the rat's lung tissue. To judge the success of the model, the experiment observed phenolsulfonphthalein elimating sputum, morphological alternations and expression of MUC5AC in the rat's lung tissue.Methods:Forty-eight rats were divided into three groups randomly: normal group, model group and treatment group. Daily exposure SD rats to inhale SO2 and cold freeze established rat model of phlegm blocking. (1)Test of phenolsulfonphthalein elimating sputum was tested. (2) The rat's lung tissue section stained by HE was observed to study the morphological alternations. (3) The expression of MUC5AC was tested with the method of immunohistochemistry. (4)After the rats were killed, their lungs were taken out for bronchi-alveli fluid and AM was cultivated. The tested cells were primary cultured alveolar macrophages. The membrance fluidity of AM was observed by DPH fluorescent probe method. (5) The macrophages were cultivated. Their ultrastructural changes were obsereved under transmission electron microscopy (TEM). (6)The secretion of nitrogen oxide(NO)and tumornecrosis factor- α (TNF-α ) in the alveolar macrophages of rats were measured. NO content was detected with enzyme methods to and TNF- a was detected with enzyme-linked immunoadsordent assay(ELISA).(7)The level of CD11a,CD11b, CD18,CD54,CD62L on AM and white cells in blood was tested with flow cytometry. (8) By means of reverse transcription polymerase chain reaction (RT-PCR), C0X-2mRNA in lung tissues in rats was measured. (9) The expression of NF- κB in lung tissues was tested with the method of immunohistochemistry.Results:(l) The volume of elimating sputum of the model group is higher than that of the normal group (P<0.01). The level of the treatment group obviously decreased, which was very markedly different from that of the model group (P<0.05). (2) There are the features of goblet cells hyperplasia in model group. The above-mentioned lung damage could be rehabilitated through treating of Chinese pharmacy. (3) The expression of MUC5AC is higher than that of the normal group(P<0.05). The level of the treatment group obviously decreased, which was very different from that of the model group (P<0.05). (4) The membrance fluidity of PAM obviously decreased in the model group, which was very markedly different from that of the normal group (P<0.05). (5) The number of mitochondria, lysosomes, smooth and rough endoplasmic reticula show the increase in model group. Especially the increase of lysosomal number in alveolar macrophages of ultrastructure are more obvious in model group that of the normal group. (6)The secretion of nitrogen oxide(NO)and tumornecrosis factor - a (TNF- a ) in the alveolar macrophages of the model group is higher than that of the normal group(P<0.01).The level of the treatment group obviously decreased, which was very markedly different from that of the model group (PO.01). (7) The levels of CD11a, CD11b, CD18, CD54, CD62L of the model group are higher than that of the normal group(P<0.01).The level of the treatment group obviously decreased, which was very markedly different from that of the model group (P<0.01). (8)The expression of NF- κB in the tissue of the bronchial wall of the model group is higher than that of the normal group (P<0.01). The percentage of cells nuclear staining of the treatment group obviously decreased, which was very markedly different from that of the model group (PO.01). (9) The expression of C0X-2mRNA in lung tissues is higher than that of the normal group (P<0.01). The level of the treatment group obviously decreased, which was very markedly different from that of the model group (PO.01).Conclusi...