The Regulatory Mechanism Of The Cell Cycle Distribution Of T Lymphocytes In Patients With Asthma Attack And The Effect Of Pulmonary Surfactant On It

Objective: (1) to investigate the cell cycle distribution and the expression of cell cycle regulatory proteins(CCRP) in T , CD4+T and CD8+T lymphocytes from patients with asthma attack, elucidate the regulatory mechanism of T lymphocytes excess activation and proliferation. (2) By cell culture technique, the CD4+T CD8+T lymphocytes obtained from peripheral blood of 20 patients with asthma attack and 20 healthy subjects were incubated with pulmonary surfactant(PS) and Phosphate-buffered saline(PBS), and the cell cycle distribution and the expression of CCRP in CD4+T CD8+T lymphocytes were anasyzed by flow cytometry in order to assess the effect of PS on the cell cycle distribution and the expression of CCRP in T lymphocyte subsets. To disclose the therapy significance of CCRP target intervention in patients with asthma attack.Methods: (1) peripheral blood was obtained from 30 patients with asthma attack and 30 normal subjects, peripheral blood mononuclear cell(PBMC)were separated by Ficoll-hypaque separation liquid, T lymphocytes were separated by nelyon cotton, CD4+T,CD8+T lymphocytes were separated by complement dependent cytotoxic. (2) DNA stained with Propidium Iodide was anasyzed by flow cytometry, according to the amount of DNA, the cell cycle distribution of T, CD4+T and CD8+T lymphocytes were analyzed using the EXPO2 DNA analysis program. Using indirect immunofluorescence technique, P27klpl cyclin E cyclin A cyclin B in T, CD4+T and CD8+T lymphocytes were anasyzed by flow cytometry. By finding the difference between patients with asthma attack and normal subjects to elucidate the regulatory mechanism of T lymphocytes proliferation cycle in patients with asthma attack. (3) By cell culture technique, the CD4 CD8+T lymphocytes obtained from peripheral blood of 20 patients with asthma attack and normal subjects were incubated withPS and PBS, the cell cycle distribution and the expression of CCRP in CD4+ CD8+T lymphocytes were analysed by flow cytometry in order to assess the effect of PS on the cell cycle distribution and expression of CCRP in T lymphocyte subsets.Results: (1) The percentage of DNA synthesis S phase and cell division G2/M phases T lymphocytes in patients with acute asthma attack were significantly higher than those in normal subjects, but G0/G1 phase were significantly lower than those in normal subjects. (2) The percentage of DNA synthesis and cell division phase CD4+T lymphocytes in patients with acute asthma attack were significantly higher than those in normal subjects. (3) The percentage of DNA synthesis CD8+T lymphocytes in patients with acute asthma attack were significantly higher than those in normal subjects, but there was no statistical difference in the percentage of cell division CD8+T in asthma patients and normal subjects. (4) The positive CCRP expression level of G1/S and G2/M restriction point in T, CD4+T and CD8+T lymphocytes in patients with asthma attack was higher than that in normal subjects; The negative CCRP were significantly lower than those in normal subjects. (5) PS inhibited the positive CCRP and increased the negative CCRP expression level of G1/S and G2/M restriction point in CD4+T lymphocytes from patients with asthma attack, inhibited DNA synthesis and cell division of CD4+T lymphocytes; PS inhibited the positive CCRP and increased the negative CCRP expression level of G1/S restriction point in CD8+T lymphocytes, inhibited DNA synthesis of CD8+T lymphocytes. (6) PS play a similar role in the cell cycle distribution and expression of CCRP in CD4+T and CD8+T lymphocytes in normal subjects and patients with asthma attack, but its regulatory degree was significantly higher in patients with asthma attack than in normal subjects.Conclusions: (1) There are significant difference in cell cycle distribution and expression of CCRP in T, CD4+T and CD8+T lymphocytes in patients with asthma attack and normal subjects, the percentage of proliferation phase T lymphocytes in patients with acute asthma attack were significantly higher than those in normal subjects. (2) T...