Linxia Fan1, Xiaojun Wang2, Linlan Fan3, Qizhang Chen1, Hong Zhang4, Hui Pan5, Aixia Xu6, Hongjuan Wang1, Yang Yu7. 1. a Respiratory Department in the Cadre Ward , Gansu Provincial Hospital , Lanzhou , China. 2. b Respiratory Department , Gansu Provincial Hospital , Lanzhou , China. 3. c Medical Experimental Center of Lanzhou University , Lanzhou , China. 4. d Asthma Center of the Gansu Provincial Hospital , Lanzhou , China. 5. e Department of Internet Information , Gansu Provincial Hospital , Lanzhou , China. 6. f Department of Pharmacy , Gansu Provincial Hospital , Lanzhou , China. 7. g Department of Internal Medicine , Traditional Chinese Medicine Hospital of Lintao County , Lintao , China.
Abstract
AIM OF THE STUDY: To delineate the underlying mechanism of microRNA-145 modulate the balance of Th1/Th2 via targeting RUNX3 in asthma patients. MATERIALS AND METHODS: Peripheral blood samples were collected from asthma patients and healthy controls. CD4+ T cells were isolated and cultured. Using quantitative PCR detect, the level of microRNA-145 and RUNX3 mRNA level in the CD4+ T cells from asthma patients and healthy controls, meanwhile, western blot was used to detect the RUNX3 protein level. Th1 or Th2 related cytokines were measured by enzyme-linked immunosorbent assay. Dual-Luciferase Reporter Assay was performed to confirm the correlation between microRNA-145 and RUNX3. MicroRNA-145 mimic or inhibitor was transfected in the CD4+ T cells and the changes of RUNX3 level, Th1 or Th2 related cytokines and the percentage of Th1 and Th2 were observed after transfection. RESULTS: MicroRNA-145 level of CD4+ T cells was higher with a lower RUNX3 expression in asthma patients. There is negative correlation between microRNA-145 and RUNX3. Th2 hyperactivity and Th1 deficiency was detected in the CD4+ T cells of asthma patients. Dual-Luciferase Reporter Assay has shown that RUNX3 is a target of microRNA. Up-regulation or down-regulation of miR-145 level caused RUNX3 expression changes in CD4+ T cells and influence the related cytokines. Inhibition of microRNA-145 may reverse the imbalance of Th1/Th2 in asthma patients. CONCLUSION: MicroRNA-145 could regulate the balance of Th1/Th2 through targeting the RUNX3 in asthma patients. MicroRNA-145 and RUNX3 may be used as biomarkers or targets in the diagnosis or therapy of asthma.
AIM OF THE STUDY: To delineate the underlying mechanism of microRNA-145 modulate the balance of Th1/Th2 via targeting RUNX3 in asthmapatients. MATERIALS AND METHODS: Peripheral blood samples were collected from asthmapatients and healthy controls. CD4+ T cells were isolated and cultured. Using quantitative PCR detect, the level of microRNA-145 and RUNX3 mRNA level in the CD4+ T cells from asthmapatients and healthy controls, meanwhile, western blot was used to detect the RUNX3 protein level. Th1 or Th2 related cytokines were measured by enzyme-linked immunosorbent assay. Dual-Luciferase Reporter Assay was performed to confirm the correlation between microRNA-145 and RUNX3. MicroRNA-145 mimic or inhibitor was transfected in the CD4+ T cells and the changes of RUNX3 level, Th1 or Th2 related cytokines and the percentage of Th1 and Th2 were observed after transfection. RESULTS: MicroRNA-145 level of CD4+ T cells was higher with a lower RUNX3 expression in asthmapatients. There is negative correlation between microRNA-145 and RUNX3. Th2 hyperactivity and Th1 deficiency was detected in the CD4+ T cells of asthmapatients. Dual-Luciferase Reporter Assay has shown that RUNX3 is a target of microRNA. Up-regulation or down-regulation of miR-145 level caused RUNX3 expression changes in CD4+ T cells and influence the related cytokines. Inhibition of microRNA-145 may reverse the imbalance of Th1/Th2 in asthmapatients. CONCLUSION: MicroRNA-145 could regulate the balance of Th1/Th2 through targeting the RUNX3 in asthmapatients. MicroRNA-145 and RUNX3 may be used as biomarkers or targets in the diagnosis or therapy of asthma.
Authors: Madhur D Shastri; Wai Chin Chong; Kamal Dua; Gregory M Peterson; Rahul P Patel; Malik Q Mahmood; Murtaza Tambuwala; Dinesh K Chellappan; Nicole G Hansbro; Shakti D Shukla; Philip M Hansbro Journal: Inflammopharmacology Date: 2020-11-05 Impact factor: 4.473