当归多糖抑制PI3K/AKT途径阻碍平滑肌细胞增殖迁移

Angelica polysaccharides inhibits the proliferation and migration of vascular smooth muscle cells by suppressing PI3K/AKT pathway

探讨当归多糖对人血管平滑肌细胞(hVSMCs)增殖,侵袭和迁移的调节作用。制备低、中、高剂量(100、200、400 mg/kg)当归多糖血清,使用不同浓度当归多糖血清预处理hVSMCs,确定当归多糖血清最佳的预处理剂量。之后,设置空白组、AngⅡ(1 ng/mL)诱导+空白血清组、HB-EGF(1 ng/mL)诱导+空白血清组、AngⅡ+当归多糖血清组、HB-EGF+当归多糖组、HB-EGF+当归多糖血清组、AngⅡ+当归多糖血清组+PI3K/AKT激动剂(IGF-1,10μM)组和HB-EGF+当归多糖组+IGF-1组。结果表明,100 mg/kg和200 mg/kg的当归多糖血清预处理对VSMCs不会产生细胞毒性,因此,使用100 mg/kg(A0)当归多糖血清用于后续实验。AngⅡ或HB-EGF可刺激hVSMC增殖,侵袭和迁移,并能提高迁移相关蛋白MMP2和MMP9的表达水平。当归多糖血清预处理可显著抑制这些过程。另外,当归多糖血清预处理降低了AngⅡ或HB-EGF诱导的hVSMCs中PI3K和AKT的磷酸化水平。综上说明当归多糖经抑制PI3K/AKT途径阻碍了AngⅡ/HB-EGF诱导的hVSMCs增殖、侵袭和迁移。

To investigate the regulatory effects of Angelica polysaccharides(AP)on proliferation,invasion and migration of vascular smooth muscle cells(VSMCs)stimulated by angiotensinⅡ(AngⅡ)and heparin-binding epidermal growth factor(HB-EGF).Human vascular smooth muscle cells(hVSMCs)were pretreated with different concentrations of AP serum.Subsequently,AngⅡor HB-EGF was used to induce hVSMC proliferation and migration.The experiment was divided into the following groups:Blank control group,AngⅡ(1 ng/mL)+control serum group,HB-EGF(1 ng/mL)+control serum group,AngⅡ+AP serum group,HB-EGF+AP serum group,HB-EGF+AP serum group,AngⅡ+AP+PI3K/AKT activator(IGF-1,10μM)group and HB-EGF+AP serum+IGF-1 group.Cell proliferation was detected by CCK-8.Cell invasion and migration were tested by Transwell and Wound Healing assay,respectively.The expression levels of MMP2,MMP9,PI3K,p-PI3K,AKT,and p-AKT were detected by Western blot.The result showed that pretreatment with 100 mg/kg and 200 mg/kg AP serum had no cytotoxicity to VSMCs.Thus,100 mg/kg AP serum was used for subsequent experiments(A0).AngⅡor HB-EGF significantly stimulated the proliferation,invasion and migration of hVSMCs.Meanwhile,AngⅡor HB-EGF improved the expression levels of migration-related proteins MMP2 and MMP9.Of note,pretreatment of AP serum dramatically inhibited these processes.In addition,AP serum pretreatment reduced phosphorylation of PI3K and AKT in AngⅡor HB-EGF-induced hVSMCs.Co-treatment of IGF-1 reversed the inhibitory effect of AP serum on hVSMC proliferation,invasion and migration.In conclusion,AP serum can inhibit the proliferation and migration of hVSMCs induced by AngⅡor HB-EGF through inhibiting the activity of PI3K/AKT signaling pathways.