| Objective: To investigate intervention of arsenic trioxide on airway epithelial cells express T cell maturation associated protein(MAL) and Signal Transducer and Activator of Transcription 6(STAT6) and its molecular mechanisms.Methods: 32 male SD rats were divided into normal control group, asthma group and arsenic trioxide group, normal serum preparation, asthma, serum and serum containing arsenic trioxide, the rat airway epithelial cells(Rat Trachea Epithelial, RTE) into normal the control group, asthma group trioxide group, inhibitor AG490 group(hereinafter referred to as AG490 group), arsenic trioxide serum + asthma serum group. Were co-cultured with RTE, in Western blotting test(WB): signal transducer and activator of transcription 6(STAT6), eosinophil chemokines(Eotaxin) and expression of MAL in RTE; Using RT-PCR to detect the expression m RNA of STAT6, Eotaxin, MAL in RTE.Results: HE staining showed: normal rat airway and alveolar without infiltration around; rat alveolar bronchial asthma and eosinophilic cytoplasm around a lot inflammatory cells, lymphocytic infiltration, partial airway epithelium, thickening of airway smooth muscle, basement membrane thickening, airway wall thickening, airway stenosis.Western blot analysis: STAT6 protein OD values asthma group was significantly higher than the control group there was a significant difference(f = 315.74, p= 0.004); AG490 group STAT6 protein OD value of the minimum, significantly lower than the other four groups were There was a significant(f= 3245.67, p= 0.001); OD value of arsenic trioxide containing serum + asthma serum group STAT6 protein was significantly lower than that of asthma group, the difference was significant(f = 152.91,p = 0.006); arsenic trioxide group than normal OD value of the control group STAT6 protein, the difference was significant(f= 110.66,p= 0.07) in each group OD values STAT6 were: normal control group 0.57 ± 0.02, asthma group 0.73 ± 0.01, arsenic trioxide group 0.46 ± 0.03, AG490 group, 0.06 ± 0.01, arsenic trioxide containing serum + asthma serum group 0.64 ± 0.02.Eotaxin protein OD values asthma group than in the control group was significantly higher, the difference was significant(f= 3164.05, p = 0.001); OD values of arsenic trioxide containing serum + asthma serum Eotaxin protein was significantly lower than the asthma group, the difference was significant sex(f = 1224.65, p = 0.003); OD value Eotaxin protein similar to the other two groups, the difference was not significant(f = 7.35, p= 0.06). Eotaxin OD values of each group were: normal control group, 0.26 ± 0.02, asthma group 1.15 ± 0.03, arsenic trioxide group, 0.92 ± 0.01, AG490 group, 0.93 ± 0.03, arsenic trioxide serum containing serum + asthma serum group 0.67 ± 0.02.MAL protein OD values asthma group than in the control group was significantly lower, the difference was significant(f = 953.93, p= 0.003); OD values of arsenic trioxide containing serum + asthma serum group MAL protein was significantly higher than that in the asthma group, the difference was significant sex(f = 177.96, p= 0.006); OD values of serum containing arsenic trioxide + asthma serum protein MAL slightly lower than the normal control group, the difference was significant(f = 162.58,p = 0.006). OD values of each group were MAL protein: normal control group 1.08 ± 0.06, asthma group 0.24 ± 0.03, arsenic trioxide group 0.20 ± 0.03, AG490 group 0.84 ± 0.05, arsenic trioxide containing serum + asthma serogroup 0.46 ± 0.03.RT-PCR analysis: m RNA STAT6 relative gray value determination: asthma group relative gray value of STAT6 m RNA was significantly higher than normal control group, the difference was significant(f= 374.56, p = 0.005); asthma group STAT6 m RNA relative gray value was significantly higher than that AG490 group, the difference was significant(f= 109.25, p= 0.007); relative gray value asthma group than arsenic trioxide STAT6 m RNA containing serum + asthma patients was significantly increased, the difference was significant(f= 76.71, p= 0.008). STAT6 of m RNA relative gray value of each group were as follows: normal control group: 0.40 ± 0.007, asthma group: 0.95 ± 0.08, arsenic trioxide containing serum group 0.48 ± 0.06, AG490 group, 0.54 ± 0.06, arsenic trioxide containing serum + asthma serum group 0.78 ± 0.08.Eotaxin m RNA relative gray value of the measurement results: the relative gray value of the asthma group Eotaxin m RNA was significantly higher than normal control group, the difference was significant(f= 325.87,p = 0.005); asthmatic group compared with arsenic trioxide group Eotaxin m RNA relative gray value was significantly increased, the difference was significant(f= 321.476, p= 0.005); asthma group and arsenic trioxide serum containing serum + asthma serum group compared Eotaxin m RNA relative gray value was significantly increased, the difference was significant(f = 35.54, p= 0.008); normal control group and arsenic trioxide serum containing serum + asthma serum group compared with the difference was significant(f = 230.70,p= 0.005). Eotaxin m RNA in each group relative to the gray values were: normal control group: 0.44 ± 0.06, asthma group: 0.87 ± 0.07, arsenic trioxide containing serum group 0.61 ± 0.17, AG490 group, 0.53 ± 0.15, arsenic trioxide containing serum + asthma serogroup 0.52 ± 0.12.MAL m RNA relative gray value of the measurement results: m RNA asthma group MAL relative gray value was significantly lower than the normal control group, the difference was significant(f = 300.26, p= 0.004); arsenic trioxide group of MAL m RAN relative gray value than the the normal control group was not statistically significant(f = 1155.61, p=0.003); serum + m RNA asthmatic serum drug containing arsenic trioxide relative gray value was significantly higher than the asthma group, the difference was significant(f =158.05,p=0.007); arsenic trioxide containing serum + asthma serum group m RNA relative gray value than those without the normal control group was significant(f = 2.12, p= 0.92). MAL of m RNA relative gray value of each group were as follows: normal control group: 0.82 ± 0.07, asthma group: 0.53 ± 0.10, arsenic trioxide containing serum group 0.51 ± 0.05, AG490 group 0.79 ± 0.17, arsenic trioxide containing serum + asthma serum group 0.78 ± 0.06.Conclusions: 1. Asthmatic serum can induced expression of Eotaxin and STAT6 in airway epithelial cells, MAL low expression. 2. Arsenic trioxide can inhibit the expression of the Eotaxin in airway epithelial cells, and increase the expression of MAL, and its mechanism may be related to inhibition of JAK-STAT6 signaling pathway... |
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