陈皮总黄酮干预血管平滑肌细胞糖胺聚糖代谢的机制研究

Study on the Effects of Chenpi Total Flavonoids on GAGs Metabolism in Vascular Smooth Muscle Cells

目的研究陈皮总黄酮(CPTF)干预血管平滑肌细胞(VSMC)糖胺聚糖(GAGs)代谢的作用机制。方法MOVAS细胞分为正常组、模型组、CPTF组(100μg·mL-1);采用氧化低密度脂蛋白(ox-LDL)诱导建立VSMC增殖模型。采用硫酸咔唑法测定GAGs总量;以HPLC法检测半乳糖胺含量;以流式细胞仪检测成纤维细胞生长因子-硫酸乙酰肝素-成纤维细胞生长因子受体1c(bFGF-HS-FGFR1c)三元复合物的荧光信号;高通量测序检测GAGs转录组基因表达。结果与正常组比较,模型组的GAGs、硫酸软骨素(CS)含量增多(P <0.05),HS含量降低(P <0.05),Cspg5(调节CS合成)、Hs3st3b1(调节HS、肝素合成)基因表达明显上调,B3gnt3(调节鞘糖脂生物合成-乳酸和新乳酸系列)、Has3(调节透明质酸合成)、Fut1(调节鞘糖脂生物合成-globo系列)基因表达下调。与模型组比较,CPTF组的GAGs、CS含量降低(P <0.01),HS含量增多(P <0.01),B3gnt3、Has3、Fut1、Hs3st3b1基因表达上调,Cspg5基因表达明显下调。结论 CPTF可能通过调节Cspg5基因的表达及HS的合成来干预VSMC的GAGs代谢,从而发挥一定的抗动脉粥样硬化的作用。

Objective To study the mechanism of Chenpi total flavonoids (CPTF) on the metabolism of glycosaminoglycans (GAGs) in vascular smooth muscle cells (VSMC). Methods VSMC were cultured in vitro and divided into normal group, model group, and CPTF group (100 μg · mL^- 1);VSMC proliferation model was established by ox-LDL induction. Total amount of GAGs was measured by carbazole sulfate method. The fluorescence signal of fibroblast growth factor-heparan sulfate-fibroblast growth factor receptor 1c (bFGF-HS-FGFR1c) ternary complex was detected by flow cytometry. The content of galactosamine was detected by HPLC. Gene expression level of GAGs in transcriptome was detected by high-throughput sequencing. Results Compared with the normal group, GAGs and CS content in the model group increased (P < 0.05);HS content decreased (P < 0.05);the genes expression of Cspg5 (regulating CS synthesis),Hs3st3b1 (regulating HS,heparin synthesis) were up-regulated significantly, and the genes expression of B3gnt3 (regulating glycosphingolipid biosynthesis-lactic acid and new lactic acid series),Has3 (regulating HA synthesis),Fut1 (regulating glycosphingolipid biosynthesis-globo series) were down-regulated. Compared with the model group, the contents of GAGs and CS in CPTF group decreased (P < 0.01);the content of HS increased (P < 0.01);the genes expression of B3gnt3,Has3,Fut1,Hs3st3b1 were up-regulated, and the gene expression of Cspg5 was down-regulated significantly. Conclusion CPTF may interfere with the metabolism of GAGs in VSMC by regulating the expression of Cspg5 gene and the synthesis of HS, thereby play a role in anti-atherosclerosis.