| Objective:To observe the effect of Cistanche deserticola polysaccharide on the expression of beta defensin 2,sIGA,IL-2,IL-6 and T cell subsets in tracheotomy intubated rats,and to explore the effect of Cistanche deserticola polysaccharide on pulmonary immune function in tracheotomy intubated rats.Method:(1)54 healthy male SD rats were randomly divided into blank control group,endotracheal intubation model non-drug group and endotracheal intubation indwelling Cistanche polysaccharide drug group,with 18 rats in each group.According to the 1st,3rd and 7th day after modeling,they were divided into three subgroups,6 rats at each time point.(2)Rat models of tracheotomy intubation were established in all groups except blank control group.Six hours after the successful establishment of the model,the blank control group and the non-drug group of endotracheal intubation indwelling model were given the same amount of saline,while the drug group of endotracheal intubation indwelling Cistanche polysaccharide was given Cistanche polysaccharide aqueous solution,once a day,3 ml each time.(3)At 24,72 and 168 hours after the establishment of the model,6 rats in each group were selected for anesthesia,and the pathological changes of lung tissue were observed under high power light microscopy.The expression of beta-defensin-2(rBD2)in lung tissue,the level of secretory immunoglobulin A(sIgA)in alveolar lavage fluid,the content of IL-2 and IL-6 in peripheral blood and the proportion of T cell subsets CD3~+,CD4~+,CD8~+were detected.Result:(1)Establishment of the rat model of tracheotomy intubation and indwelling:The rats in the blank control group were in good mental state,breathing smoothly without abnormal sound,fur smooth and lustrous,drinking water freely and eating normally,and body weight increased in a normal range.Rats in the non-drug group had abnormal mental state,and were prone to over-stress response to external sound.There were abnormal alertness and over-sensitivity.There are many secretions in the mouth and nose,the breathing frequency is accelerated and the depth becomes shallow,there may be irregular breathing,sometimes accompanied by phlegm ringing,there are varying degrees of cheek scratching,catheterization,restlessness,shortness of breath and so on,and the above symptoms are aggravated with the passage of time.The fur is colourless,or even erect,with a bow and back,and is lazy to move.The model Cistanche deserticola polysaccharides treatment group had good mental state,and the other symptoms were milder than those of the non-treatment group,and the improvement of the above symptoms was more obvious with the passage of time.(2)Histopathological changes of lung tissue in each group:observation of lung tissue under high power light microscopy,the lung tissue of rats in blank control group was intact without pathological changes;the lung tissue of rats in model non-drug group showed pathological changes such as alveolar wall loss,infiltration of inflammatory cells in alveoli and bronchus,intracellular and interstitial hemorrhage,and with the prolongation of time,the alveolar wall became heavily thicker and a large number of neutrophils appeared.Cell infiltration,severe papillary protrusion of bronchial wall,a large number of inflammatory cells infiltration in bronchus;compared with the model non-drug group,the pulmonary pathological changes and inflammatory cell infiltration in the model Cistanche deserticola polysaccharide group were significantly improved,with varying degrees,and gradually improved over time.(3)The expression of rBD2 in lung tissues of each group:There was no significant difference in the relative expression of rBD2 in lung tissues of rats in blank control group in three time periods.Compared with the blank control group,the expression of rBD2 in the lung tissue of the model group increased significantly at 24h,then began to decline,and decreased significantly at 72h.The degree of reduction increased with time,and decreased to the lowest level at168h(P<0.01).The expression level of Cistanche deserticola polysaccharide decreased at 24h,then increased gradually,and reached the highest value at168h,which was higher than that of blank control group(all P=0.000).(4)Levels of sIgA in alveolar lavage fluid in each group:The comparison of three time periods in blank control group was all P>0.05.Compared with the blank control group,the sIgA content of alveolar lavage fluid in tracheotomy intubation model group was significantly higher than that in normal rats at 24h,decreased at 72h,and decreased significantly at 168h.The content of sIgA in alveolar lavage fluid of Cistanche deserticola polysaccharide treated group increased significantly at 24h,reached the highest level at 72h,and remained higher at 168h than that of blank control group and model group.(5)The levels of IL-2 and IL-6 in peripheral blood of rats in each group:The levels of IL-2 and IL-6 in peripheral blood of rats in blank control group showed no difference in three time periods.Compared with the blank control group,the level of IL-2 in peripheral blood of rats in the model non-drug group increased briefly at 72h,but there was no significant change in the rest of the time(P>0.05).The expression of IL-6 increased significantly at 24 hours(P<0.01),decreased slightly at 72 hours and continued to rise at 168 hours.After intervention with Cistanche deserticola polysaccharide,the level of IL-2 in peripheral blood of tracheotomy intubated rats increased significantly at 24h,and the degree of improvement was proportional to the time of administration,and was much higher than the blank control group(P<0.01).The content of IL-6 decreased significantly at 24h and reached the lowest level at 168h(P<0.01).The decreasing degree was gradually obvious with the prolongation of medication time(P<0.01).(6)The percentage of CD3~+,CD4~+,CD8~+in T lymphocyte subsets of each group:There was no difference in the percentage of CD3~+,CD4~+,CD8~+in the blank control group.The percentage of CD3~+in the non-drug group decreased significantly at 24h,increased slightly at 72h,but was much lower than that in the blank control group,and there was no significant change thereafter.In model Cistanche deserticola polysaccharide group,it decreased at 24h,rapidly returned to normal level at 72h,and continued to rise at 168h(P=0.000<0.01).There was no difference in the percentage of CD4~+in the model group in three time periods,but the ratio of CD4~+in the three time periods was lower than that in the blank control group(P=0.000<0.01).The expression of Cistanche deserticola polysaccharides in the model group increased significantly at 72h,and increased gradually with time,reaching the highest level at 168h.There was no difference in the percentage of CD8~+between the blank control group and the model non-drug group,and there was no difference in the blank control group in three time periods.The percentage of CD8~+in the model group increased at 72h,and was still significantly higher at 168h than that in the blank control group(P=0.000<0.01).The model Cistanche deserticola polysaccharide group decreased at 24h,and continued to play a decreasing role over time.Conclusion:(1)In the early stage of tracheotomy,the lung is stimulated to produce inflammation.With the release of inflammatory factors,the immune function of the lung is impaired.(2)Cistanche deserticola polysaccharide can directly enhance immune function by improving the imbalance of T cell subsets and stimulating the secretion of IL-2.At the same time,it can also reduce the release of pro-inflammatory factor IL-6 to alleviate the inflammatory reaction in the lungs of pneumotomy rats,which is conducive to the repair of local immune function in the lungs. |
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