The Effect And Mechanism Of Chaipu Decoction On Regulating IL-5、FasL Expression In Airway Epithelium Cells By Up-regulating The Mal Expression

Objective: To clarify the molecular mechanism of Chaipu decoction regulating the expressions of IL-5 and Fas L on airway epithelial cells through up-regulating the expression of MAL.Method : 32 healthy male SD rats,weight( 250±10) g,were assigned randomly into two groups, with 16 rats in each group.Rats in one group were assigned randomly into Chaipu decoction group and saline group,with 8 rats in each group,After treatment, blood was collected from rat heart, and serum was prepared. Rats in the other group was taken to establish a mice asthmatic model was established by OVA and asthmatic serum was prepared, the lung tissue was obtained and stained by HE.Rat tracheal epithelial cells were devided into groups: normal group(A), asthma group(B), pre-Chaipu decoction group(C), U0126 group(D) and Chaipu decoction group(E).Normal group to be added to 3m L of normal serum;asthma group to be added to 2m L asthma serum and 1m L of normal serum; pre-Chaipu decoction group to be added to the Chaipu decoction serum1 m L,two hours later,added asthma serum 2m L;U0126 group to be added 10μL of U0126 and 2m L asthma serum and 1m L of normal serum, Chaipu decoction group to be added to 1m L Chaipu decoction serum and 2m L asthma serum, after 24 hours of incubation, cells were extracted protein, RNA, the culture medium was collected. RT-PCR to detect airway epithelial cells MAL, IL-5, Fas L m RNA expression. Western blot detected the MAL, Fas L, p-ERK1 / 2 protein in airway epithelial cells express. The IL-5 concentration in supernatant was detected by ELISA.Result :1、The general condition of rats and the result of HE staining.1)The general condition of rats in asthma group:the rats became spiritual malaise, irritable, restless, action slowly,hair changed withered and yellow, breathing became deep and urgent with abdominal contractions. the above situation have not appeared in other groups.2) The pathologic changes of rat lung tissue by HE staining:In control group,bronchial mucous membrane presented continuous structure,no inflammatory cell infiltration,pulmonary alveoli intact wall.Bronchiolar dilatation and wall thickening,the bronchial lumen narrowing,Most alveolar damage, fusion, alveolar space enlarged, alveolar interval fracture can be seen in the asthma group.2 、 Western blot to measure the Fas L, MAL, P-ERK1 / 2 protein expressionMAL、Fas L protein was weakest in asthma group, the differences was significant(p<0.05) compared with Chaipu decoction group and normal group; MAL protein expression in the normal group was the strongest; the expression of IL-5 protein in the asthma group stronger than the Chaipu decoction group and the normal group, the difference was statistically significant(p<0.05); Chaipu decoction group and the U0126 group expression of Fas L protein higher than the asthma group(P <0.05), the difference was significant(p<0.05); P-ERK1/2 protein expression less in U0126 group, and the strongest in asthma group, the difference was statistically significant(p<0.05); Chaipu decoction group expression P-ERK1/2 protein is weaker,and comparing with asthma group the difference was meaningful(p<0.05), but still higher than the normal group and U0126 group, the difference was significance(p<0.05). MAL protein expression of Chaipu decoction group and UO126 group was not significantly different(p>0.05).Chaipu decoction group expression of MAL have no significantly different compared with pre-Chaipu decoction group(p> 0.05).3、 Secretion of IL-5 measured by ELISAThe Expression of IL-5 concentration in the asthma group was the highest, compared with Chaipu decoction group and U0126 group the difference is statistically significant(p<0.05); the IL-5 expression in Chaipu decoction group is higher than the normal group, the difference is significant(p<0.05), but it was lower than U0126 group(p<0.05); but compared with the pre-Chaipu decoction group difference was not significant(p> 0.05).4、RT-PCR to measure the expression levels of Fas L, MAL, IL-5m RNA in the cell.Fas L, MALm RNA expression weakest in the asthmatic group, compared with Chaipu decoction group, U0126 group and the normal group was statistically significant(p <0.05); expression of MALm RNA in Chaipu decoction group compared with the pre-Chaipu decoction group, U0126 group the difference was not significant(p> 0.05); expression of Fas Lm RNA in Chaipu decoction group was high than U0126 group, the difference was statistically significant(p<0.05). IL-5m RNA expression strongest in asthmatic group was statistically significant(p<0.05) difference compared with the rest of the groups;Chaipu decoction group expression IL-5m RNA higher than normal group, the difference was significant(p <0.05). The expression of IL-5m RNA in U0126 was higher than Chaipu decoction group, the two groups was statistically significant(P <0.05).Conclusion:1、Asthmatic secrum can down-regulate the expression of MAL and Fas L,and up-regulate IL-5 expression in airway epithelial cells,The mechanism may be related to activation of ERK.2 、 Chaipu decoction decrease IL-5 expression and increase Fas L expression in airway epithelial cells by up-regulating MAL expression,the mechanism may related to inhibition of ERK.