| Objective1. To observe the damage in rat tracheal mucociliary and lung tissue pathological change after intratracheal instillation with fluconazole injection.2.To research the expression and significance of TNF-α protein in lung tissue after intratracheal administration.3.Discussion the possible mechanisms of injury in tracheal mucociliary and lung tissue after intratracheal administration,while observing the intervention effect of ambroxol throug intraperitoneal injection on the above injury. MethodsRandomly divided the 90 clean healthy male SD rats into three groups: normal control group(N), fluconazole group(F), fluconazole + ambroxol group(FA group),each group with 30. N group without any treatment. In group F and FA group,inject the fluconazole slowly(0.425mg/kg) through tracheal puncture to intratracheal administration rat model.while intraperitoneally inject ambroxol(70mg / kg) in FA group 6 hours before intratracheal instillation of fluconazole injection.They were sacrificed respectively with five rats at six different time points( wo hours,one day, two days, three days, five days, seven days).Then take the lower trachea to observe he damage in rat tracheal mucociliary by the scanning electron microscopy.Take the right deputy lung lobe to Judge the pathological change after HE staining and detect lung tissue TNF-α protein immunohistochemical method.Results1.Scanning electron microscopy showed that in the N group tracheal cilia arrangement was neat, compact,without edema, necrosis and lodging.Wile in group F and group FA at each time point,the tracheal mucociliary has varying degrees of damage, the injury reached the peak in the first five days and was significantly improved in the fifth day 7 day.But in the group F cilia damage area was up to 98% while the FA group was 68%. There were statistically significant difference among the three groups at six time points after pairwise comparisons(P <0.01).2.HE staining of lung tissue showed that more sever shedding and swelling in the bronchi and bronchioles epithelial cells,more inflammatory cells and mucus exudation in the bronchi cavity,increase of erythrocytes cells and inflammatory cells infiltration in the alveolar and alveolar septa, more obvious inflammatory cell infiltration in perivascular compared with group N and group FA.3.In group N there was a small amount of TNF-α protein expression on the bronchial epithelial cells by immunohistochemistry. In group F after 2 hours the staining was the most shallow, but increased gradually in 1th day to 3th day.The 5th day the coloring was deepest.At the six time points of group FA the TNF-α protein staining scores were higher than group F and group N, the difference statistically significant(P <0.01). Conclusions1.After a local intratracheal injection of fluconazole injection, there are different degrees of damage on rat tracheal cilia and lung mucosa.The extent of damage on the fifth day reach peake but on day 7th the damage recovered gradually,which hinted the damage is partially reversible, but further tests was on the way to prove if it is completely reversible.2.After the intratracheal local injection the increase of lung tissue inflammatory cytokines TNF-α protein expression tips TNF-α may be involved in the process of this damage.3. Ambroxol can relieve the injury of intratracheal injection fluconazole on trachea mucosa and lung tissue,which may be related to the variety lung protective effect of ambroxol.We speculate that one of the specific mechanism may be related to the inhibition release of inflammatory mediators, decreased lung tissue levels of TNF-α expression. |
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