氧化型低密度脂蛋白对人血管平滑肌细胞透明质酸表达的影响及机制

目的观察氧化型低密度脂蛋白(ox-LDL)对人主动脉血管平滑肌细胞透明质酸(HA)合成的影响,并初步探究其分子机制。方法体外培养人主动脉血管平滑肌细胞T/G HAVSMC,使用不同浓度(10、25、50、75、100 mg/L)ox-LDL对T/G HAVSMC细胞进行干预,使用CCK-8法检测细胞存活率,并进行分组分析。采用浓度为25和50 mg/L的ox-LDL干预T/G HAVSMC细胞48 h,并设置天然LDL(native-LDL,N-LDL)组(50 mg/L的N-LDL)和对照组,使用HPLC法测定HA含量,使用实时定量PCR检测透明质酸合成酶2(HAS2)和透明质酸合成酶3(HAS3)的mRNA表达,使用Western blot法检测凝集素样氧化型低密度脂蛋白受体1(LOX-1)、低密度脂蛋白受体相关蛋白1(LRP-1)、氧化脂蛋白的清道夫受体(SR-PSOX)以及脂肪酸转位酶(CD36)的表达。结果低于100 mg/L的ox-LDL对T/G HAVSMC细胞无明显细胞毒性。25 mg/L和50 mg/L ox-LDL组HA含量、HAS2和HAS3的mRNA表达量明显高于N-LDL组和对照组(P0.05),N-LDL组与对照组组间差异无显著性(P0.05)。25 mg/L和50 mg/L ox-LDL组LOX-1表达明显高于N-LDL组和对照组(P0.05),而LRP-1、SR-PSOX和CD36表达无明显变化(P0.05)。结论 ox-LDL能够诱导人主动脉平滑肌细胞中HA的合成,其机制可能与结合LOX-1有关。

Effect of oxidized low density lipoprotein on hyaluronic acid expression in human vascular smooth muscle cells and its mechanism

Aim To investigate the effect of oxidized low-density lipoprotein (ox-LDL) on the synthesis of hyaluronic acid (HA) in human aortic vascular smooth muscle cells (HAVSMC), and explore its molecular mechanism. Methods The T/G HAVSMC were cultured in vitro with different concentration of ox-LDL(0,5, 0,5, 100 mg/L), the CCK-8 method was used to measure the cell proliferation and grouping analysis was carried out. T/G HAVSMC were treated with ox-LDL at concentrations of 25 and 50 mg/L for 48 hours. Natural LDL (N-LDL) group (50 mg/L N-LDL) and control group were set up. HPLC was used to determine the HA content, real-time quantitative PCR was used to detect the mRNA level of hyaluronic acid synthetase 2 (HAS2) and hyaluronic acid synthase 3 (HAS3), Western blot was used to detect the protein level of lectin-like oxidized low density lipoprotein recepter-1 (LOX-1), low-density lipoprotein receptor-related protein-1 (LRP), scavenger receptor for phosphatidylserine and oxidized lipoprotein (SR-PSOX) and cluster of differentiation 36 (CD36). Results After 48 h intervention, ox-LDL had no significant cytotoxicity on T/G HAVSMC cells. After 25 and 50 mg/L ox-LDL intervention for 48 h, the content of HA were significantly higher than those in the N-LDL group and the control group(P<0.05), the mRNA expression levels of HAS2 and HAS3 were significantly higher than those in the N-LDL group and the control group(P<0.05), and there was no significant difference between the N-LDL group and the control group(P>0.05). The expression of LOX-1 in 25 mg/L ox-LDL group and 50 mg/L ox-LDL group was significantly higher than that in the N-LDL group and the control group(P<0.05), while the expression of LRP-1, SR-PSOX and CD36 showed no significant change(P>0.05). Conclusion ox-LDL could induce the synthesis of HA in human aortic smooth muscle cells, and its mechanism may be related to the combination of LOX-1.