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目的探讨p38 MAPK通路是否通过激活内质网应激及凋亡参与高糖所致血管平滑肌细胞(vascular smooth muscular cell,VSMCs)钙化。方法大鼠VSMCs分为对照组、高糖组(35μmol/L D-葡萄糖)、内质网应激抑制剂4-苯基丁酸(4-PBA)组、p38 MAPK通路抑制剂SB203580组、β-磷酸甘油组、4-PBA+高糖组、SB203580+高糖组、β-磷酸甘油+高糖组,分别用比色法、o-cresolphthalein法和Western blot测定碱性磷酸酶(ALP)活性、钙含量和骨分化转录因子(Runx2和Osterix)表达。结果 D-葡萄糖处理3、7d上调VSMCs ALP活性、钙含量和骨分化标志蛋白表达;SB203580下调ALP活性、钙含量和骨分化转录因子表达,而4-PBA抑制高糖引起VSMCs内质网应激及凋亡。结论 p38 MAPK通路通过激活内质网应激和凋亡可能是高糖诱导VSMCs钙化的重要机制。
Objective To investigate whether p38 MAPK pathway is involved in the calcification of vascular smooth muscle cells (VSMCs) induced by high glucose by activating endoplasmic reticulum stress and apoptosis. Methods Rat VSMCs were divided into control group, high glucose group (35μmol / L D-glucose), endoplasmic reticulum stress inhibitor 4-phenylbutyric acid (4-PBA) group, p38 MAPK pathway inhibitor SB203580 group, - phosphoglycerate group, 4-PBA + high glucose group, SB203580 + high glucose group and β-phosphoglycerate + high glucose group. The activities of ALP and calcium were determined by colorimetric method and o-cresolphthalein method, Content and expression of osteogenic transcription factors (Runx2 and Osterix). Results D-glucose up-regulated the ALP activity, calcium content and osteogenic marker protein expression of VSMCs. SB203580 down-regulated ALP activity, calcium content and osteogenic transcription factor expression, while 4-PBA inhibited high glucose induced VSMCs endoplasmic reticulum stress And apoptosis. Conclusion p38 MAPK pathway may be an important mechanism of high glucose-induced calcification of VSMCs through activation of endoplasmic reticulum stress and apoptosis.