SARAF In The Functional Regulation Of Asthmatic Airway Smooth Muscle

Objective Store-operated calcium channel(SOCC)participates in asthmatic airway smooth muscle(ASM)contraction,and hypertrophy.SOCE-associated regulatory factor(SARAF)is a newly identified molecule,which mainly regulates slow calciumdependent inactivation(SCDI)of SOCC.This study first explored the role of SARAF in asthmatic airway inflammation,airway hyperresponsiveness,and airway remodeling,and secondly investigate the role of SARAF in the proliferation,migration,and regulation the activity of SOCC in human airway smooth muscle cells(ASMCs).Method 1)Acute and chronic OVA-sensitized asthmatic mouse models were established and divided into acute control group(A-PBS group),acute asthma group(A-OVA group),acute asthma lentivirus transfection control group(A-NC group),acute asthma lentivirus transfected SARAF overexpression group(A-SARAF group),chronic control group(C-PBS group),chronic asthma group(C-OVA group),chronic asthma lentivirus transfection control group(C-NC group),chronic asthma lentivirus transfected SARAF overexpression group(C-SARAF group),5 mice in each group.Airway inflammation,airway hyperresponsiveness(AHR)and airway remodeling were measured in each group.The relative mRNA expression of SARAF in the ASM of each group was evaluated by RT-PCR.The expression of IgE in BALF of each group was evaluated by ELISA.2)Cultrued human ASMCs were divided into control group(PBS group),siRNA control group(NC group)and SARAF siRNA group(SARAF RNAi group).The transfection efficiency was tested by RT-PCR.Cell proliferation was measured by CCK-8 test.Cell migration was assessed using Scratch-Wound assay.The activity of SOCC was tested by live cell workstation.Result 1)A mouse model of acute asthma was successfully established.The relative mRNA expression of SARAF in the ASM of A-OVA group was significantly lower than that of A-PBS group,airway inflammation and RI of A-OVA group were significantly higher than that of A-PBS goup.While the relative mRNA expression of SARAF in the ASM of A-SARAF group was significantly higher than that of the other three groups,and airway inflammation and RI of A-SARAF were significantly lower than that of A-OVA group and A-NC group.2)A mouse model of chronic asthma was successfully established.The relative mRNA expression of SARAF in ASM of C-OVA group was significantly lower than that of C-PBS group,while C-SARAF group was significantly higher than that of other three groups.Goblet cell hyperplasia,the collagen deposition and smooth muscle hyperplasia of C-OVA group,C-NC group were significantly higher than that of C-PBS group,while C-SARAF group was significantly lower than that of C-NC group.3)Human airway smooth muscle cells were successfully cultrued in vitro.Downregulation of SARAF in ASMCs significantly increased cell proliferation rate,migration rate and enhanced the activity of SOCC.Conclusion SARAF may inhibit airway inflammation,airway resistance and airway remodeling in asthmatic mice.SARAF may inhibit cell proliferation,migration and the activity of SOCC.SARAF may be a new target for asthma treatment research.