CAJ | 학술논문

目的：探讨内质网(endoplasmic reticulum,ER)应激在高糖引起的血管平滑肌细胞(vascular smooth muscle cells,VSMCs)钙化中的作用与机制。方法35 mmol/L D-葡萄糖(高糖)处理 VSMCs,模拟糖尿病状态,观察高糖能否引起 VSMCs 的 ER 应激和 VSMCs 表型转化(收缩型转变为成骨样细胞),观察 ER 应激的抑制剂对VSMCs 钙化的效应,采用比色法、o-cresolphthalein 法和 western blot 观察碱性磷酸酶活性、钙沉积和骨分化转录因子(Runx2)等指标。结果应用35 mmol/L D-葡萄糖分别处理 VSMCs 3 d 或7 d,可引起 VSMCs 的 ER 应激蛋白表达上调、碱性磷酸酶活性增加、钙沉积和骨分化标志蛋白增多；苯基丁酸(4-phenylbutyric acid,4-PBA)预处理在抑制 VSMCs 的 ER 应激的同时,能阻断高糖所致 VSMCs 钙化(表现为碱性磷酸酶活性、钙沉积和骨分化标志蛋白下降)。结论高糖能激活 VSMCs 的 ER 应激,继而引起 VSMCs 凋亡和钙化,提示 ER 应激在高糖引起的VSMCs 钙化中起着重要作用。
목적：탐토내질망(endoplasmic reticulum,ER)응격재고당인기적혈관평활기세포(vascular smooth muscle cells,VSMCs)개화중적작용여궤제。방법35 mmol/L D-포도당(고당)처리 VSMCs,모의당뇨병상태,관찰고당능부인기 VSMCs 적 ER 응격화 VSMCs 표형전화(수축형전변위성골양세포),관찰 ER 응격적억제제대VSMCs 개화적효응,채용비색법、o-cresolphthalein 법화 western blot 관찰감성린산매활성、개침적화골분화전록인자(Runx2)등지표。결과응용35 mmol/L D-포도당분별처리 VSMCs 3 d 혹7 d,가인기 VSMCs 적 ER 응격단백표체상조、감성린산매활성증가、개침적화골분화표지단백증다；분기정산(4-phenylbutyric acid,4-PBA)예처리재억제 VSMCs 적 ER 응격적동시,능조단고당소치 VSMCs 개화(표현위감성린산매활성、개침적화골분화표지단백하강)。결론고당능격활 VSMCs 적 ER 응격,계이인기 VSMCs 조망화개화,제시 ER 응격재고당인기적VSMCs 개화중기착중요작용。
Objective To investigate the role and mechanism of endoplasmic reticulum (ER)stress response in calcification of vascular smooth muscle cells (VSMCs)induced by high glucose.Methods VSMCs mimicked Diabetes was treated with 35 mmol/L D-glucose to observe whether high glucose could induce ER stress response and calcification of VSMCs.Effects of ER stress inhibitor on calcification of VSMCs was observed.ALP activity, calcium content and osteogenic markers expression were tested using colorimetric assay,o-cresolphthalein method and Western blot analysis.Results Treating VSMCs with 35 mmol/L D-glucose for 3d or 7d could induce ER stress and osteoblastic differentiation of VSMCs (ALP activity,calcium content and osteogenic markers expression increase).4-phenylbutyric acid (4-PBA)pre-treatment could inhibit ER stress of VSMCs and high glucose-elicited vascular SMCs calcification.Conclusion High glucose can activate ER stress,apoptosis and osteoblastic differentiation of VSMCs which demonstrates the important role of ER stress in VSMCs calcification induced by high glucose.