广东医学
廣東醫學
엄동의학
GUNAGDONG MEDICAL JOURNAL
2015年
7期
981-984
,共4页
周龙霞%赵丽%陈久凯%杨安宁%刘现梅%王艳华%姜怡邓%曹军
週龍霞%趙麗%陳久凱%楊安寧%劉現梅%王豔華%薑怡鄧%曹軍
주룡하%조려%진구개%양안저%류현매%왕염화%강이산%조군
高同型半胱氨酸血症%内质网应激%葡萄糖调节蛋白78%C/EBP环磷酸腺苷反应元件结合转录因子同源蛋白%胱冬酶-12
高同型半胱氨痠血癥%內質網應激%葡萄糖調節蛋白78%C/EBP環燐痠腺苷反應元件結閤轉錄因子同源蛋白%胱鼕酶-12
고동형반광안산혈증%내질망응격%포도당조절단백78%C/EBP배린산선감반응원건결합전록인자동원단백%광동매-12
hyperhomocysteinemia%endoplasmic retculum stress%glucose-regulatedprotein78%C/EBP homolo-gousprotein%Caspase-12
目的：探讨高同型半胱氨酸血症（ HHcy）对ApoE－／－鼠心脏内质网应激（ ERS）的影响及其可能机制。方法取5周龄雄鼠12只作为正常对照组；另取5周龄雄性ApoE－／－小鼠36只，随机分为3组，ApoE－／－对照组：ApoE－／－小鼠饲以普通饲料配方饮食；高蛋氨酸组：ApoE－／－小鼠饲以高蛋氨酸饮食；干预组：ApoE－／－小鼠饲以高蛋氨酸饮食同时加0.006％叶酸和0.0004％维生素B12。喂养20周后处死，行主动脉病理切片HE染色观察主动脉是否有斑块形成；取心脏，采用实时荧光定量PCR法检测GRP78、CHOP、Caspase－12 mRNA的变化；巢式降落式甲基化特异性PCR检测GRP78和CHOP启动子区DNA甲基化水平。结果正常对照组及ApoE－／－对照组斑块不明显，高蛋氨酸组动脉粥样硬化斑块面积明显增大，主动脉内膜可见灶性病变部位隆起突入动脉腔内，干预组动脉粥样硬化斑块面积较高蛋氨酸组有所减轻。与ApoE－／－对照组比较，高蛋氨酸组GRP78、CHOP、Caspase－12 mRNA水平明显升高（P＜0.05，P＜0.01），干预组GRP78、CHOP mRNA水平较高蛋氨酸组降低（P＜0.05）。与ApoE－／－对照组比较，高蛋氨酸组GRP78、CHOP启动子区DNA甲基化水平降低（P＜0.01），干预组GRP78、CHOP甲基化水平较高蛋氨酸组有所升高（ P＜0.05）。结论 HHcy可能通过影响DNA甲基化程度引起ERS相关基因表达改变，进而导致心肌细胞凋亡，补充叶酸与维生素B12可有效缓解HHcy所引起的ERS和凋亡。
目的：探討高同型半胱氨痠血癥（ HHcy）對ApoE－／－鼠心髒內質網應激（ ERS）的影響及其可能機製。方法取5週齡雄鼠12隻作為正常對照組；另取5週齡雄性ApoE－／－小鼠36隻，隨機分為3組，ApoE－／－對照組：ApoE－／－小鼠飼以普通飼料配方飲食；高蛋氨痠組：ApoE－／－小鼠飼以高蛋氨痠飲食；榦預組：ApoE－／－小鼠飼以高蛋氨痠飲食同時加0.006％葉痠和0.0004％維生素B12。餵養20週後處死，行主動脈病理切片HE染色觀察主動脈是否有斑塊形成；取心髒，採用實時熒光定量PCR法檢測GRP78、CHOP、Caspase－12 mRNA的變化；巢式降落式甲基化特異性PCR檢測GRP78和CHOP啟動子區DNA甲基化水平。結果正常對照組及ApoE－／－對照組斑塊不明顯，高蛋氨痠組動脈粥樣硬化斑塊麵積明顯增大，主動脈內膜可見竈性病變部位隆起突入動脈腔內，榦預組動脈粥樣硬化斑塊麵積較高蛋氨痠組有所減輕。與ApoE－／－對照組比較，高蛋氨痠組GRP78、CHOP、Caspase－12 mRNA水平明顯升高（P＜0.05，P＜0.01），榦預組GRP78、CHOP mRNA水平較高蛋氨痠組降低（P＜0.05）。與ApoE－／－對照組比較，高蛋氨痠組GRP78、CHOP啟動子區DNA甲基化水平降低（P＜0.01），榦預組GRP78、CHOP甲基化水平較高蛋氨痠組有所升高（ P＜0.05）。結論 HHcy可能通過影響DNA甲基化程度引起ERS相關基因錶達改變，進而導緻心肌細胞凋亡，補充葉痠與維生素B12可有效緩解HHcy所引起的ERS和凋亡。
목적：탐토고동형반광안산혈증（ HHcy）대ApoE－／－서심장내질망응격（ ERS）적영향급기가능궤제。방법취5주령웅서12지작위정상대조조；령취5주령웅성ApoE－／－소서36지，수궤분위3조，ApoE－／－대조조：ApoE－／－소서사이보통사료배방음식；고단안산조：ApoE－／－소서사이고단안산음식；간예조：ApoE－／－소서사이고단안산음식동시가0.006％협산화0.0004％유생소B12。위양20주후처사，행주동맥병리절편HE염색관찰주동맥시부유반괴형성；취심장，채용실시형광정량PCR법검측GRP78、CHOP、Caspase－12 mRNA적변화；소식강락식갑기화특이성PCR검측GRP78화CHOP계동자구DNA갑기화수평。결과정상대조조급ApoE－／－대조조반괴불명현，고단안산조동맥죽양경화반괴면적명현증대，주동맥내막가견조성병변부위륭기돌입동맥강내，간예조동맥죽양경화반괴면적교고단안산조유소감경。여ApoE－／－대조조비교，고단안산조GRP78、CHOP、Caspase－12 mRNA수평명현승고（P＜0.05，P＜0.01），간예조GRP78、CHOP mRNA수평교고단안산조강저（P＜0.05）。여ApoE－／－대조조비교，고단안산조GRP78、CHOP계동자구DNA갑기화수평강저（P＜0.01），간예조GRP78、CHOP갑기화수평교고단안산조유소승고（ P＜0.05）。결론 HHcy가능통과영향DNA갑기화정도인기ERS상관기인표체개변，진이도치심기세포조망，보충협산여유생소B12가유효완해HHcy소인기적ERS화조망。
Objective To investigate the effects of hyperhomocysteinemia ( HHcy ) on myocardial endoplasmic reticulum stress ( ERS) in ApoE-/-mice and its possible mechanism.Methods Thirty-six 5 -week old ApoE-/-C57BL/6J mice were randomly divided into three groups, including ApoE-/-control group, ApoE-/-HHcy group and the intervention group.Twelve 5-week old normal C57BL/6J mice were served as normal group.After 20 weeks of feed-ing, the aortae were separated for HE stain and observation of aortic plaque.GRP78, CHOP and Caspase-12 mRNA lev-els were determined by qRT-PCR.The methylation status of GRP78 and CHOP were studied using the nest touch down methylation specific polymerase chain reaction ( ntMS-MSP) .Results No significant aortic plaque was observed in ei-ther normal group or ApoE-/-control group.In the ApoE-/-HHcy group, the aortic plaque areas were greater, breaking into the aortic lumina, which were alleviated in the intervention group.The mRNA levels of GRP78, CHOP and Caspase-12 in the ApoE-/-HHcy group were significantly higher than those in ApoE-/-control group(P<0.05,P<0.01), while they were significantly down-regulated in the intervention group(P<0.05,P<0.01).GRP78 and CHOP DNA methylation levels were significantly lowered in the ApoE-/-HHcy group(P<0.01,P<0.01),while significanty higher in the intervention group(P<0.05,P<0.05).Conclusion HHcy may affect Myocardial ERS via changing DNA methy-lation levels, then lead to apoptosis.Earlier folic acid and VB12 interference can effectively reduce the myocardial ERS and apoptosis.
