| Objective:To make use of LPS induced mice ARDS model and PDTC to intervene and to detect the expression of cytokines TNF-α, IL-1β, IL-8 in mice plasma and to detect the expression of CD11b/CD18 in lung tissue of mice and to study the influence of PDTC on mice ARDS cytokines TNF-α, IL-1β, IL-8and CD11b/CD18 expression,to provide further theoretical support for the protective effects of PDTC on ARDS. Methods: 54 healthy male BALB/c mice were randomly divided into the normal control group(groupN), ARDS group(group L), PDTC intervention group(group P + L), with 18 mice in each group. Group N was given intraperitoneal injection of normal saline 20ml/kg, group L was given intraperitoneal injection of LPS 20mg/kg to induce mice ARDS model, and group P+L was given intraperitoneal injection of PDTC 120mg/kg 30 minutes before LPS injection. Six rats were taken from each group at 4h, 8h, and 12 h after the modeling, mouse orbital blood was collected and ELISA technology was used to detect cytokines TNF-α, IL-1β and IL-8 expression level in the plasma; after left ventricular blood, arterial blood gas analysis and calculation oxygenation index(PaO2/FiO2); the right lung was taken to calculate W/D; the left lung was taken to make paraffin blocks and slices for HE staining. The lung tissue pathology change was observed under light microscope and immunohistochemical detection was used to detect CD11b/CD18 expression in the lung tissue. Using SPSS17.0 statistical software to deal with the experimental data, the numeric variables using mean ±standard deviation( x ±s), two random sample were using t test, multiple sets of comparing the method of analysis of variance, P￡0.05 indicates that the difference had statistical significance. Results: 1. General condition of mice from different groups: the breath of mice from group N was smooth, the spirit condition was good, and when they were grab, they moved quickly; the breath frequency of mice from group L was quick, the spirit was low, and when they were grab, they moved slowly or didn’t move; the breath of mice from group P+L was urge, the spirit was better than group L, and when they were grab, they moved quickly than group L, too. 2. Oxygenation index, Pa O2/FiO2 resultsof mice from different groups: the oxygenation index, PaO2/FiO2 of mice from group L at 4h, 8h and 12 h was evidently lower than group N and the difference had statistical significance(P￡0.05). With the prolonging of time, the oxygenation index, PaO2/FiO2 of mice from group L was lower and lower. The oxygenation index, PaO2/FiO2 of mice from group P+L at 4h, 8h and 12 h was evidently higher than group L and the difference had statistical significance(P￡0.05), but still lower than group N and the difference had statistical significance(P￡0.05). 3. Lung tissue gross sample andpathological changes in mice lung tissue of different groups: the gross sample of lung tissue of mice from group N was in pink,and observed under light microscope after HE staining, there was no evident pathological changes in the mice lung tissue; the gross sample volume of lung tissue of mice from group L was large and in dark red, hemorrhagic spots can be seen on the surface and liquid leakage can be seen in the cut section, and observed under light microscope after HE staining, hemorrhage, a large number of inflammatory cells infiltration, alveolar walls fracture can be seen in the lung tissue of mice from group L; there was no evident enlargement in the gross sample volume of lung tissue of mice from group P+L, the color was in red and a small amount of bleeding can be seen on the surface, and observed under light microscope after HE staining, a small amount of hemorrhage and inflammatory cell infiltration can be seen in the lung tissue and the alveolar wall breaking condition was lighter than group L. 4. Lung wet/dry weight ratio(W/D)of mice from different groups: the mice lung tissue W/D value of group L was evidently higher than that of group N and the difference had statistical significance(P￡0.05). With the prolonging of time, the pulmonary edema was aggravating gradually; the lung tissue W/D value of group P+L was lighter than group L and the difference had statistical significance(P￡0.05), but still higher than group N and the difference had statistical significance(P￡0.05). 5. TNF-α、IL-1β、IL-8expression in mice plasma of different groups: TNF-α、IL-1β、IL-8 concentration of mice plasma from group L at 4h, 8h and 12 h was evidently higher than group N and the difference had statistical significance(P￡0.05). With the prolonging of time, TNF-α、IL-1β、IL-8expression level was higher and higher. TNF-α、IL-1β、IL-8concentration of mice plasma from group P+L at 4h, 8h and 12 h was evidently lower than group L and the difference had statistical significance(P￡0.05), but still higher than group N and the difference had statistical significance(P￡0.05). 6. CD11b/CD18 expression in mice lung tissue of different groups: Brown in the lung tissue indicates positive expression. The immunohistochemical results showed that CD11b/CD18 expression in the lung tissue of group N was few expression or almost no expression; CD11b/CD18 expression in the lung tissue of groupL was positive, significantly increased compared to group N, and with the prolonging of time, the positive expression increased gradually; CD11b/CD18 expression in the lung tissue of group P+L decreased significantlycompared to group L,but still evidently higher compared to group N. Conclusion:1. CytokinesTNF-α, IL-1β, IL-8 and adhesion molecule CD11b/CD18 expression increase significantlyin LPS induced ARDS in mice, indicating that TNF-α, IL-1β, IL-8 and CD11b/CD18 participate in the process of ARDS. 2. PDTC can improve ARDS lung injury in mice caused by LPS, and it may be related to its decreasing the expression of TNF-α, IL-1β, IL-8 and CD11b/CD18. |
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