TUDCA减轻慢性间歇性缺氧诱导的巨噬细胞MMP-9蛋白表达
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- 英文论文题名：TUDCA Attenuates Intermittent Hypoxia-Induced MMP-9 Protein Expression in Macrophage
- 论文作者：杨云云 ; 李林忆 ; 杨颂 ; 王嫕 ; 秦彦文
- 年：2016
- 作者机构：首都医科大学附属北京安贞医院北京市心肺血管疾病研究所;
- 论文关键词：阻塞性睡眠呼吸暂停综合征 ; 慢性间歇性缺氧 ; 动脉粥样硬化 ; 内质网应激 ; 基质金属蛋白酶 ; 组织蛋白酶
- 英文论文关键词：obstructive sleep apnea syndrome ; chronic intermittent hypoxia ; atherosclerosis ; endoplasmic reticulum stress ; matrix metalloproteinase ; cathepsins
- 会议召开时间：2016-05-27
- 会议录名称：中国睡眠研究会第九届学术年会汇编
- 语种：中文
- 分类号：R766
- 学会代码：OGSF
- 会议名称：中国睡眠研究会第九届学术年会
- 会议地点：中国上海
- 主办单位：中国睡眠研究会
- 学会名称：中国睡眠研究会
- 页数：3
- 文件大小：116k
- 原文格式：O
- 会议级别：全国
摘要
背景:阻塞性睡眠呼吸暂停低通气综合征(Obstructive sleep apnea-hypopnea syndrome,OSAHS)是一种全身性疾病,主要表现为打鼾、呼吸暂停、夜间反复低氧血症、高碳酸血症和睡眠结构紊乱。研究认为OSAHS是心血管疾病的独立危险因素。慢性间断缺氧是OSAHS主要的病理生理基础,能够促进动脉粥样硬化形成和斑块破裂,机制尚未明确。基质金属蛋白酶(MMP)和组织蛋白酶(Cathepsins)均可通过促进炎症和细胞外基质降解参与动脉粥样硬化的发病过程。内质网应激是动脉粥样硬化形成的机制之一。目的:本研究探讨内质网应激在慢性间歇性缺氧促进巨噬细胞(THP-1)蛋白酶表达中的作用。方法及结果:在三气培养箱中建立人巨噬细胞慢性间歇性缺氧细胞模型(氧浓度2%-21%60min一个循环),分为三组:正常氧对照组、慢性间歇性缺氧组、慢性间歇性缺氧+内质网应激抑制剂牛磺熊去氧胆酸(TUDCA)组,分别给予24h、48h、72h的慢性间歇性缺氧,以不同剂量的500umol/L TUDCA(2.5uL、5uL)干预。光学显微镜下观察细胞形态变化,MTT方法检测细胞活力,免疫印迹方法检测内质网应激相关蛋白葡萄糖调节蛋白78(GRP78)、内质网激酶样蛋白激酶(PERK)、磷酸化真核启动因子2α(P-eIF2α)、激活转录因子4(ATF4)的表达,同时检测蛋白酶MMP-9和Cathepsin S的蛋白表达。结果:慢性间歇性缺氧处理组随时间延长,在48h、72h能够显著促进MMP-9、Cathhepsin S的表达,其总表达量与对照组相比有显著提高(P<0.05);慢性间歇性缺氧在48h、72h内促进内质网应激相关蛋白GRP78、PERK、P-eIF2α、ATF4的表达,与对照组相比其表达增高显著(P<0.05);TUDCA高剂量组(5uL)能显著抑制慢性间歇性缺氧诱导的MMP-9和PERK的蛋白表达(P<0.05);结论:慢性间歇性缺氧通过内质网应激途径促进THP-1细胞MMP-9和Cathepsin S的蛋白表达。TUDCA可能通过PERK途径抑制慢性间歇性缺氧诱导的THP-1细胞MMP-9的表达,可能为OSAHS合并动脉粥样硬化的防治提供新的靶点。
Background:Obstructive sleep apnea-hypopnea syndrome(OSAHS) is a systemic disease,mainly for snoring and apnea,repeatedly hypoxemia at night,hypercapnia,and disorders of sleep structure.Studies suggest that OSAHS are independent risk factors of cardiovascular disease.Chronic intermittent hypoxia is the main pathological physiological basis of OSAHS.Chronic intermittent hypoxia promoted the formation of atherosclerosis and plaque rupture,but its mechanism is not yet clear.Matrix metalloproteinases(MMPs) and cysteine protease(Cathepsins)participate in the formation of atherosclerosis through promoting inflammation and extracellular matrix degradation.Endoplasmic reticulum stress is one of mechanism of atherosclerosis.Objective:This study is to explore the role of endoplasmic reticulum stress in chronic intermittent hypoxia induce protease expression in macrophage(THP-1).Methods and Results:in CO_2 incubator(Thermo-311) build human macrophages cells model of chronic intermittent hypoxia(oxygen concentration from 2%to 21%60min/cycle),the cells were divided into three groups:normoxic control group,chronic intermittent hypoxia group,chronic intermittent hypoxia and endoplasmic reticulum stress inhibitors(TUDCA) group,respectively give24 h,48h,72 h of chronic intermittent hypoxia,and use the same concentration(500umol/L)different doses of TUDCA(2.5uL 5uL) intervention.Optical microscope cell morphological changes,MTT method to detect the cell vitality,western blot method to detect the expression of endoplasmic reticulum stress related protein glucose-regulated protein 78(GRP78),protein kinaselike ER kinase(PERK),phosphorylation eukaryotic initiation factor 2 alpha(P-elF2alpha),activating transcription factor 4(ATF4),and the expression of MMP-9 and Cathepsis S level.Results:chronic intermittent hypoxia treatment group with the extension of the time,in 48 hour,72hour can significantly promote the expression of MMP-9,cathepsin S,its total expression has increased significantly compared with control group(p < 0.05);Chronic intermittent hypoxia in48 hour,72hour promoted the expression of endoplasmic reticulum stress related proteins GRP78,PERK,P-eif2 alpha,ATF4,its expression increased significantly compared with control group(p <0.05);TUDCA(5 ul) group significant inhibite the endoplasmic reticulum stress PERK protein expression(p < 0.05);TUDCA(5 ul) group could significantly inhibit the chronic intermittent hypoxia induced the expression of MMP-9(p < 0.05).Conclusion:Chronic intermittent hypoxia promoted the protein expression of MMP- 9 and cathepsin S through the endoplasmic reticulum stress pathway in THP-1 cells.TUDCA suppress expression of MMP-9 induced by chronic intermittent hypoxia in THP-1 cells may through inhibiting PERK pathway.This may provide a new target for the prevention and treatment of atherosclerosis combined with OSAHS.
Background:Obstructive sleep apnea-hypopnea syndrome(OSAHS) is a systemic disease,mainly for snoring and apnea,repeatedly hypoxemia at night,hypercapnia,and disorders of sleep structure.Studies suggest that OSAHS are independent risk factors of cardiovascular disease.Chronic intermittent hypoxia is the main pathological physiological basis of OSAHS.Chronic intermittent hypoxia promoted the formation of atherosclerosis and plaque rupture,but its mechanism is not yet clear.Matrix metalloproteinases(MMPs) and cysteine protease(Cathepsins)participate in the formation of atherosclerosis through promoting inflammation and extracellular matrix degradation.Endoplasmic reticulum stress is one of mechanism of atherosclerosis.Objective:This study is to explore the role of endoplasmic reticulum stress in chronic intermittent hypoxia induce protease expression in macrophage(THP-1).Methods and Results:in CO_2 incubator(Thermo-311) build human macrophages cells model of chronic intermittent hypoxia(oxygen concentration from 2%to 21%60min/cycle),the cells were divided into three groups:normoxic control group,chronic intermittent hypoxia group,chronic intermittent hypoxia and endoplasmic reticulum stress inhibitors(TUDCA) group,respectively give24 h,48h,72 h of chronic intermittent hypoxia,and use the same concentration(500umol/L)different doses of TUDCA(2.5uL 5uL) intervention.Optical microscope cell morphological changes,MTT method to detect the cell vitality,western blot method to detect the expression of endoplasmic reticulum stress related protein glucose-regulated protein 78(GRP78),protein kinaselike ER kinase(PERK),phosphorylation eukaryotic initiation factor 2 alpha(P-elF2alpha),activating transcription factor 4(ATF4),and the expression of MMP-9 and Cathepsis S level.Results:chronic intermittent hypoxia treatment group with the extension of the time,in 48 hour,72hour can significantly promote the expression of MMP-9,cathepsin S,its total expression has increased significantly compared with control group(p < 0.05);Chronic intermittent hypoxia in48 hour,72hour promoted the expression of endoplasmic reticulum stress related proteins GRP78,PERK,P-eif2 alpha,ATF4,its expression increased significantly compared with control group(p <0.05);TUDCA(5 ul) group significant inhibite the endoplasmic reticulum stress PERK protein expression(p < 0.05);TUDCA(5 ul) group could significantly inhibit the chronic intermittent hypoxia induced the expression of MMP-9(p < 0.05).Conclusion:Chronic intermittent hypoxia promoted the protein expression of MMP- 9 and cathepsin S through the endoplasmic reticulum stress pathway in THP-1 cells.TUDCA suppress expression of MMP-9 induced by chronic intermittent hypoxia in THP-1 cells may through inhibiting PERK pathway.This may provide a new target for the prevention and treatment of atherosclerosis combined with OSAHS.
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