作者机构：
[Zhang Cai-Ping] Univ South China, Coll Med, Hengyang 421001, Peoples R China.;[Lin Li-Mei; Tuo Qin-Hui; Sun Shao-Wei; Gong Yong-Zhen; Liao Duan-Fang] Hunan Univ Chinese Med, State Key Lab Chinese Med Powder & Med Innovat Hu, Div Stem Cell Regulat & Applicat, Changsha 410205, Hunan, Peoples R China.;[Zhang Cai-Ping; Zheng Xing; Sun Shao-Wei; Ou Lu; Lei Xiao-Yong] Univ South China, Coll Pharm & Biol Sci, Hengyang 421001, Peoples R China.;[Tuo Qin-Hui; Gong Yong-Zhen; Liao Duan-Fang] Hunan Univ China, Sinoluxembourg Cooperat Res Ctr Chinese Med, Med, Changsha 410208, Hunan, Peoples R China.
通讯机构：
[Liao Duan-Fang] H;Hunan Univ Chinese Med, State Key Lab Chinese Med Powder & Med Innovat Hu, Div Stem Cell Regulat & Applicat, Changsha 410205, Hunan, Peoples R China.
摘要：
Aim: To investigate the mechanisms of anti-atherosclerotic action of ezetimibe in rat vascular smooth muscle cells (VSMCs) in vitro. Methods: VSMCs of SD rats were cultured in the presence of Chol:MβCD (10 μg/mL) for 72 h, and intracellular lipid droplets and cholesterol levels were evaluated using Oil Red O staining, HPLC and Enzymatic Fluorescence Assay, respectively. The expression of caveolin-1，sterol response element-binding protein-1 (SREBP-1) and ERK1/2 were analyzed using Western blot assays. Translocation of SREBP-1 and ERK1/2 was detected with immunofluorescence. Results: Treatment with Chol: MβCD dramatically increased the cellular levels of total cholesterol (TC)，cholesterol ester (CE) and free cholesterol (FC) in VSMCs, which led to the formation of foam cells. Furthermore, Chol:MβCD treatment significantly decreased the expression of caveolin-1, and stimulated the expression and nuclear translocation of SREBP-1 in VSMCs. Co-treatment with ezetimibe (3 μmol/L) significantly decreased the cellular levels of TC, CE and FC, which was accompanied by elevation of caveolin-1 expression, and by a reduction of SREBP-1 expression and nuclear translocation. Co-treatment with ezetimibe dose-dependently decreased the expression of phosphor-ERK1/2 (p-ERK1/2) in VSMCs. The ERK1/2 inhibitor PD98059 (50 μmol/L) altered the cholesterol level and the expression of p-ERK1/2, SREBP-1 and caveolin-1 in the same manner as ezetimibe did. Conclusion: Ezetimibe suppresses cholesterol accumulation in rat VSMCs in vitro by regulating SREBP-1 and caveolin-1 expression, possibly via the MAPK signaling pathway.
摘要：
Aims: This study aims to examine the possible associations between high density lipoprotein (HDL) subclass distribution and APOA5-1131T>C polymorphism in hypertriglyceridemia. Methods: The distribution of HDL subclasses was quantified by 2-dimensional electrophoresis in conjunction with immunodetection method. The APOA5-1131T>C polymorphism was identified in 95 hypertriglyceridemic (HTG) patients and 102 healthy subjects by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Results: The APOA5-1131C (C) allele frequency was higher in the HTG group than in the control group. Plasma triglycerides (TG) were significantly higher and apoA5 was significantly lower in patients with the C allele when compared to patients with the APOA5-1131T (T) allele, even more dramatically so in the APOA5-1131CC homozygote. In both the HTG group and the control group, the frequency of the C allele was positively correlated with levels of TG, total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and apolipoprotein B100 (apoB100), and negatively correlated with levels of high density lipoprotein cholesterol (HDL-C), apolipoprotein A1 (apoA1) and apolipoprotein A5 (apoA5) (P < 0.001). In all subjects, the frequency of the C allele was positively correlated with the level of small-sized HDL (pre beta(1)-HDL and HDL3a), and negatively correlated with levels of HDL2a and HDL2b. Conclusion: Changes in HDL subclass distributions in HTG may be related to the APOA5-1131T>C polymorphism. This polymorphism leads to a general shift towards smaller-sized HDL. (c) 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
作者机构：
[Song lan] Cent S Univ, Xiangya Sch Med, Dept Pathophysiol, Changsha 410008, Hunan, Peoples R China.;[Xu Zhao-jun] Hunan Univ Tradit Chinese Med, Cardiothorac Surg Affiliated Hosp 1, Changsha 41007, Hunan, Peoples R China.;[Zhang Cai-ping; Tian ying] Nanhua Univ, Coll Life Sci, Dept Biochem & Mol Biol, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Song, L ] ;Cent S Univ, Xiangya Sch Med, Dept Pathophysiol, 110 Xiangya Rd, Changsha 410008, Hunan, Peoples R China.
关键词：
fibroblast;adrenaline;bFGF;TGF-beta 1
摘要：
Adrenaline has been shown to modulate proliferation of mouse fibroblasts, adventitial fibroblasts and synovial B (fibroblasts-like) cells. However, little is known about the response of cultured human hypertrophic scar fibroblasts to adrenaline. In this study, we investigated cell proliferation and involved mechanisms in hypertrophic scar fibroblasts in response to adrenaline. Population doubling time (PDT) assay and MTT assay were performed to determine the cell proliferation and cell viability, respectively. The expression of bFGF and TGF-beta 1 was measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). The results showed that adrenaline inhibited proliferation of normal and hypertrophic scar fibroblasts in a dose-dependent manner. Moreover, adrenaline up-regulated the expression of bFGF and down-regulated the expression of TGF-beta 1 in normal and hypertrophic scar fibroblasts. Interestingly incubation with the a receptor antagonist regitine indicated that adrenaline mediated inhibition of cell proliferation and regulation of TGF-beta 1 and bFGF in cultured normal and hypertrophic scar fibroblasts were mediated by the a receptor. These studies suggest that adrenaline inhibits proliferation and alters the expression of TGF-beta 1 and bFGF in human hypertrophic scar fibroblast involving an a receptor mediated pathway.
