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Migration of VSMC and SPC by Expression of Sdf-1αin VSMC on the Effect of ox-LDL

Author: ZhangPeng
Tutor: RuanQiuRong
School: Huazhong University of Science and Technology
Course: Pathology and Pathophysiology
Keywords: Vascular Smooth Muscle Cell Smooth Muscle Progenitor Cells Stromal Cell Derived Factor-1a Oxidized Low Density Lipoprotein Mouse Atherosclerosis
CLC: R363
Type: Master's thesis
Year: 2010
Downloads: 7
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Objective1. To explore the effect of oxidized low density lipoprotein (ox-LDL) on expression of stromal cell derived factor-1α(SDF-1α) in mouse vascular smooth muscle cell (VSMC).2. Compare migration of vascular smooth muscle cell and smooth muscle progenitor cell (SPC) by the effect of stromal cell derived factor-1αand conditioned medium of VSMC.Methods1. Culture VSMC of mouse by tissue—piece inoculation and identify VSMC by the method of immunochistochemistry (IHC).2. VSMC was incubated with different concentration/time of ox-LDL, protein of SDF-1αwas revealed by the method of immunochistochemistry and immunosorbent assay (ELISA) and mRNA of SDF-1αwas revealed by transcription-polymerase chain reaction (RT-PCR), Observed the effect of concentration-time.3. Experiment of migration: Put SDF-1αin the basic room of the Transwell chamber; Put VSMC and SPC in the upside room of the Transwell chamber; Put Transwell chamber in 37℃for 6 hours; Fixed by alcohol; Count the number of cell by Microscope; Reveal effect of migration in VSMC and SPC by SDF-1αand conditioned medium of VSMC.Result1. Purity of the cultured VSMC was about 90%.2. SDF-1αwas constitutionally expressed in mouse VSMC. SDF-1αin mouse VSMC was changed by ox-LDL with 0~60ng/ml and the time of 24h,protein of SDF-1α’s expression was peaked at 45ng/ml and was more 1.4times than basal expression. SDF-1αin mouse VSMC was changed by time of 0~48h and the ox-LDL of 45ng/ml, protein of SDF-1α’s expression was peaked at 24h and was more 1.5 times than basal expression, mRNA of SDF-1α’s expression was peaked at 10h and was more 3.2 times than basal expression.3. Effect of migration in VSMC and SPC is obvious by the effect of SDF-1αand conditioned medium of VSMC and the effect of migration in VSMC is more obvious than the effect of migration in SPC.Conclusion1. SDF-1αin mouse VSMC was increased by ox-LDL.2. The effect of migration in VSMC is more obvious than the effect of migration in SPC by the effect of SDF-1αand conditioned medium of VSMC.

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