Effects Of Artemisinin On Th17/Treg Cytokines In Peripheral Blood Of Patients With Bronchial Asthma

Objective:Through the change of the concentration in the peripheral blood of patients with bronchial asthma IL-10, IL-17, IL-25, IL-35 detection of artemisinin before and after the intervention, according to the effects of the of different periods in patients with asthma in the peripheral blood of these factor density change to explore effects of artemisinin on bronchial asthma pathogenesis and treatment.Methods:1.Randomly collected 20 healthy people as healthy control group, as well as 20 cases of remission and acute attack of asthma patients as experimental group, each person's early morning fasting venous blood 5ml in the heparin anticoagulant tube;2.Ficoll density gradient centrifugation was used to separate the single nuclear cell solution(PBMCs), and the RPMI-1640 culture solution was used to adjust the PBMCs density to 2 * 107 /ml;3.The PBMCs solution of the healthy group, the remission phase and the acute episode asthma patients were respectively added into the 24 hole culture plate, and the PBMCs of the same sample were put into two holes, each hole and 1ml;4.Tealthy group the two holes were added in RPMI-1640 medium 10 ul and cell stimulating agent 2ul, and remission period of acute exacerbation of two holes were added in RPMI-1640 medium 10 ul and cell stimulating agent 2ul, artemisinin(0.16umol/ml) 10 ul and cell stimulating agent 2ul;5.Then the culture plates are placed in 37 C and 5% CO2 incubator in cultured 24h;6.The collection plate in PBMCs from liquid culture were placed in ready to ATC 3000r / min, and centrifuged for 20 min, leaving PBMCs culture supernatant in 1.5ml Eppendorf tube, placed in refrigerator- 80 DEG C to preserve the reserve;7.Finally, thaw- 80 degrees refrigerator spare PBMCs culture supernatant, strictly in accordance with the enzyme linked immunosorbent assay(ELISA) kit to operate detected OD value of all specimens, and then use the corresponding ELISA software to calculate the concentration of each inflammation cytokine, IL-10, IL-17, IL-25, IL-35, and comparative analysis of the data.Results:1.1640 intervention, concentration of Th17 cells can secrete cytokines IL-17, IL-25 in remission group and acute attack period group than in healthy group increased(P < 0.05),and acute phase increased more significantly(P < 0.05); concentration of Treg cells secrete cytokines IL-10, IL-35 in remission group and acute attack period group than in healthy group decreased(P < 0.05), acute phase decreases with the degree of greater(P < 0.05);2. Artemisinin intervention to ease the group with acute stage group in Th17 cells can secrete cytokines IL-17, IL-25 concentration decreased(P < 0.05), acute attack period group and remission group are decreased more(P < 0.05); and Treg cells secrete cytokines IL-10, IL-35 concentrated A.(P < 0.05), the effect of artemisinin intervention of acute phase cytokines concentration changes than in remission phase is more significant, the difference is statistically significant(P < 0.05).Conclusion:1. Artemisinin can regulate the concentration changes of Th17 and Treg cells of cytokines in supernatants of PBMCs in patients with asthma and reduce the concentration of Th17 cells can secrete cytokines IL-17, IL-25, elevated concentrations of Treg cells secrete cytokines IL-10, IL-35, namely artemisinin can significantly improve bronchial asthma patients and in the peripheral blood of inflammation cytokine IL-10, IL-17, IL-25,IL-35 concentration index;2. The mechanism of artemisinin in the treatment of bronchial asthma may be related to the regulation of the balance between IL-10, IL-17, IL-25 and IL-35 concentrations in Th17/Treg cells in vivo, to improve the immune imbalance of Th17/Treg cells and achieve the purpose of treatment of asthma.