ABSTRACT
Objective To investigate the effects of Qingzao Jiufei Tang(QJD)and its decomposing agent on the levels of TNF-α,INF-γ levels of TNF-α of Qingo, P1 and AQP5 in lung tissue of mice infected with Mycoplasma pneu-moniae(MP),and to clarify its molecular anti-infective mechanism. Methods One hundred-forty-four SPF grade BABL/c mice were randomly divided into the normal group(group A),model group(group B),QJD group(group C),QJD de-composition agent I group(group D)、QJD decomposition group II Group(group E)and azithromycin group(group F), with 24 rats in each group. Besides the group A,the other 5 groups of mice were treated with MP infection. After the mod-eling,the mice were given corresponding drugs by gastric gavage,and samples were obtained on the days 3,7,10,14 af-ter the model was established. The lung tissue sections were examined by histopathology, and the degrees of inflammation in lung tissues in the mice were evaluated,and the lung index and the ratio of dry and wet lung weight ratio in the mice were calculated. The levels of MPN372 and P1 genes were determined by qPCR assay. The serum TNF-α and INF-γ levels in the mice were assessed by enzyme linked immunosorbent assay(ELISA). The expression of AQP5 protein was detected by Westernblot. Results After MP infection, the pathological examination revealed thickening of alveolar septum and bronchioles,and extensive inflammatory cell infiltration in the lung tissues. The lung index was increased and the ratio of dry and wet lung weight(P<0.05). TNF- α and INF-γ cedd and was increased, and reached the peak on the seventh day. The expression of AQP5 protein showed a downward tendency,and began to gradually increase on the 14th day. Com-pared with the group B,the expression levels of MPN372,P1 and TNF-α in the group D were down-regulated,and the ex-pression levels of INF-γ levein the group E were up-regulated from the 7th day. Conclusions QJD can control pulmonary inflammation in mice after MP infection. The decrease of production of MP toxin MPN372 and the expression of adhesion protein P1,the up-regulation of expression of INF-γ and AQP5 proteins,and down-regulation of TNF-α expression are one of the mechanisms of its action.
ABSTRACT
Objective To observe the effect of Qingzao-Jiufei decoction and its composition on the levels of IL-10,TNF-α, CD4+cells, CD8+T cells and the change of CD4+/CD8+ratio in the serum to explore the mechanism of anti-MP infection. Methods The 144 SPF grade BALB/c mice were randomly divided into the normal group, the model, the whole decoction group, the composition Ⅰ group, the composition Ⅱgroup with the method of random digital table, and the azithromycin group and every group had 24 mice. The following five groups were treated with infection of MP model. After successful modeling, the rats in the whole decoction group were treated by gavage with 1 g/ml Qingzao-Jiufei decoction. The rats in the composition Ⅰ group were treated by gavage with 0.61 g/ml composition Ⅰ decoction. The rats in the composition Ⅱ group were treated by gavage with 0.39 g/ml composition Ⅱ decoction. And the three groups were administered for 14 days. The rats in the azithromycin group were treated by gavage with azithromycin suspension once a day. After three days, the drug was stopped. Each group was infected on 3,7,10,14 day. Then mice were sacrificed, with the method of taking the eye of the mice to take blood. The level of IL-10, TNF-α in the mice blood with the method of ELISA were detected. The level of CD4+, CD8+T in the rinsing solution of mice spleen with the method of FCM and the CD4+/CD8+ ratio were detected. Results Compared with the model group, the expressions of CD4+T cells (24.50 ± 1.41 vs. 22.08 ± 1.99) and IL-10 (18.15 ± 0.36 vs. 8.75 ± 0.16) in the whole decoction group significantly increased. The expressions of CD8+T cells (7.29 ± 1.23 vs. 9.13 ± 1.14) and TNF-α (28.32 ± 1.90 vs. 37.97 ± 1.71) significantly decreased (P<0.05), and the level of CD4+/CD8+ratio increased. In the composition Ⅰ group, the expressions of CD8+T cells (7.50 ± 1.45 vs. 9.13 ± 1.14) and TNF-α(33.48 ± 1.08 vs. 37.97 ± 1.71) significantly decreased (P<0.05). In the composition Ⅱ group, the expressions of CD4+T cells (23.63 ± 1.10 vs. 22.08 ± 1.99) and IL-10 (17.82 ± 0.63 vs. 8.75 ± 0.16) significantly increased (P<0.05). Conclusions The mechanism of Qingzao-Jiufei decoction on anti-MP infection may be related to the regulation of the cellular immune.
ABSTRACT
Objective To compare the changes in TLR4-NF-κB signaling pathway in infant and adult mice infected with influenza virus, and to provide experimental evidence for the study of immunopathological mechanism in pediatric respiratory virus susceptibility. Methods Immunohistochemistry and RT-PCR were applied to detect the expressions of lung TLR4 and NF-κB P65 mRNA and proteins in the infant and adult mice, and to compare the changes in TLR4-NF-κB P65 signaling pathway after infection with influenza virus.Results (1) The infant model group showed the strongest expression of TLR4 protein in the lung tissue, compared with that in the normal group and adult model group showing significant differences (P<0.05).(2) The expression of NF-κB P65 protein in the lung tissue was strongest in the infant model group, and it was gradually increased over time, showing a significant difference between each time point and the next time point (P<0.05).(3) The infant model group showed the strongest expression of TLR4 mRNA in lung tissue, significantly higher than that in the normal and adult model groups (P<0.05).(4) The expression of NF-κB P65 mRNA in the lung tissue was highest in the infant model group, and significantly higher than that in the normal and the adult model groups ( P<0.05) , and it was gradually increased with the time.Conclusions The over-activation of TLR4-NF-κB P65 signaling pathway may be one of the immunopathological mechanisms of serious injury in the lung tissue in infant rats.