| Objective:To investigate influence of DUSUQING (DSQ) of Traditional Chinese Medicine Presciptions on JAK/STAT signal transduction pathway in the lung tissue of bacterial pneumonia rat models with phlegm-heat syndrome and to reveal the mechanism of DSQ in treating bacterial pneumonia with phlegm-heat syndrome.Methods:Wistar rats were randomly divided into normal group,pneumonia group,pneumonia with phlegm-heat syndrome group,positive control group and DSQ treated group. To make bacterial pneumonia model by trachea intubation, the phlegm-heat symptom was made on the basis of the bacterial pneumonia model using wind-heat stimulating under the heating circumstance. The rats in positive control and DSQ treated group were respectively perfused Qingjin huatantang soup with Sangbaipi soup, andDSQ hydrosolvent (amount to raw medicine lg/ml) in 7th day, the dose respectively was 5.607g·kg-1·d-1 and 2.8g·kg-1·d-1, one time each day, for five days. Normal group, pneumonia group and phlegm-heat syndrome group were syringed with the same volume NS into rats'stomach once a day, for five days.The pathological histological changes of lung tissue were observed through the Light microscopy. The expressions levels of IL-1,IL-6,IL-10,TNF-a,p-JAK2,p-STATl,p-STAT3 and SOCS3 proteins in lung tissue were analyzed by immunohistochemistry staining.The expression levels of IL-1mRNA, IL-10mRNA and SOCS3 mRNA was measured by RT-PCR.Results:1 Changes of leukocyte counts and neutrophil ratios in peripheral blood in each group:The count of WBC and the percentage of PMN in peripheral blood in pneumonia group and pneumonia with phlegm-heat syndrome group were more evidently than in normal group (P<0.01). The count of WBC and the percentage of PMN in pneumonia with phlegm-heat syndrome group was higher than in pneumonia group(P<0.01,P<0.05). The count of WBC and the percentage of PMN in positive control group and DSQ treated group were lower than in pneumonia with phlegm-heat syndrome group(P<0.01, P>0.05). Compared with the positive control group,the count of WBC and the percentage of PMN in DSQ group were reduced,but there was no statistical significance(P>0.05).2 Changes of pathological histological of lung tissues in each group:There were normal lung tissue structure in normal group.The exudation of phlogistic was seen in bronchial lumens and alveoli spaces,and there were hyperaemia,edemas, inflammatory cell infiltration in the lung tissue in pneumonia group. Compared with the pneumonia group, the pathological injury of lung tissue in pneumonia with phlegm-heat syndrome group were particularly severe. Slight inflammatory reaction was found in positive control group and DSQ treated group, and the inflammatory damage in DSQ treated group was much lighter than that in positive control group.3 Changes of the expressions of IL-1βand IL-1βmRNA in lung tissue in each group:The expressions of IL-1βand IL-1βmRNA in pneumonia group and in pneumonia with phlegm-heat syndrome group were markedly higher than that in normal control group(P<0.01),And in pneumonia with phlegm-heat syndrome group the expressions were even higher, but there was no statistical significance(P>0.05). Compared with pneumonia with phlegm-heat syndrome group,the expressions of IL-1βand IL-1βmRNA were markedly weakened in positive control group and DSQ group (P<0.01), Compared with the positive control group,the expressions of IL-1βand IL-1βmRNA in DSQ group were particularly reduced (P<0.05).4 Changes of the expressions of TNF in lung tissue in each group:The expressions of TNF in pneumonia group and in pneumonia with phlegm-heat syndrome group were markedly higher than that in normal control group(P<0.01),And in pneumonia with phlegm-heat syndrome group the expressions were significantly even higher(P<0.01). Compared with pneumonia with phlegm-heat syndrome group,the expressions of TNF were markedly weakened in positive control group and DSQ group (P<0.01), Compared with the positive control group,the expressions of TNF in DSQ group were particularly reduced (P<0.05).5 Changes of the expressions of IL-6 in lung tissue in each group:Compared with normal group,the expressions of IL-6 in pneumonia group and in pneumonia with phlegm-heat syndrome group were markedly higher than it (P<0.01),And in pneumonia with phlegm-heat syndrome group the expressions were significantly even higher(P<0.05). there were lower expressions in positive control group and DSQ group than that in pneumonia with phlegm-heat syndrome group(P<0.01),and cmpared with the positive control group,the expressions of IL-6 in DSQ group were particularly reduced (P<0.05).6 Changes of the expressions of IL-10 and IL-lOmRNA in lung tissue in each group:The expressions of IL-10 and IL-lOmRNA in pneumonia group and in pneumonia with phlegm-heat syndrome group were markedly higher than that in normal control group(P<0.01),And in pneumonia with phlegm-heat syndrome group the expressions were even higher (P<0.05). Compared with pneumonia with phlegm-heat syndrome group,the expressions of IL-10 and IL-10mRNA were markedly enhanced in DSQ group (P<0.05).7 Changes of the expressions of p-JAK2,p-STATl,p-STAT3 in lung tissue in each group:The expressions of p-JAK2,p-STATl,p-STAT3 in pneumonia group and in pneumonia with phlegm-heat syndrome group were markedly higher than that in normal control group(P<0.01),And in pneumonia with phlegm-heat syndrome group the expressions were significantly even higher(P<0.05). Compared with pneumonia with phlegm-heat syndrome group,the expressions of p-JAK2,p-STATl,p-STAT3 proteins were markedly weakened in positive control group and DSQ group(P<0.01). Compared with the positive control group,the expressions of p-JAK2,p-STATl,p-STAT3 in DSQ group were significantly reduced (P<0.05).8 Changes of the expressions of SOCS3 protein and SOCS3 mRNA in lung tissue in each group:The expressions of SOCS3 protein and SOCS3 mRNA in pneumonia group and in pneumonia with phlegm-heat syndrome group were markedly higher than that in normal control group(P<0.01),And in pneumonia with phlegm-heat syndrome group the expressions were significantly even higher(P<0.05).Compared with pneumonia with phlegm-heat syndrome group,the expressions of SOCS3 protein and SOCS3mRNA were enhanced in positive control group and DSQ group (P<0.01).Compared with the positive control group,the expressions level of SOCS3 protein and SOCS3 mRNA in DSQ group were even higher, but there was no statistical significance(P>0.05).Conclusion:1 The bacterial pneumonia with phlegm-heat syndrome model was successfully made on the basis of the bacterial pneumonia model with wind-heat stimulating under the heating circumstance. The disease and syndrome model are some differences in white blood cells reaction, the expressions of inflammatory cytokines and the JAK/ STAT signal transduction pathway from the single disease model。The differences reflect the pathophysiological features of its own.2 The unbalance between proinflammatory and anti-inflammatory factor and JAK /STAT signal transduction pathway is involved in the development of bacterial pneumonia with phlegm-heat syndrome.3 DSQ has significant protective effects against the inflammatory injury of lung tissue,and there were some possible mechanisms to explain it.First, DSQ may directly inhibit excessive release of proinflammatory cytokine such as TNF-a, IL-1βand IL-6.Secondly,it can promote IL-10 secretion to regulate the balance between pro-and anti-inflammatory responses.Thirdly,it may interrupt JAK/STAT transduction pathway and reducing genes expressions of the downstream immunity cytokines. Finaly, it can be involved in regulating the expression of SOCS3-a negative feedback factor in JAK/STAT signaling pathway, in turn affect the pathway, but the mechanism needs further exploration.
|
PDF Full Text Request