山核桃叶总黄酮及其单体化合物对血管紧张素Ⅱ诱导的血管平滑肌细胞增殖和迁移的抑制作用

Inhibitory Effects of Total Flavonoids and Pure Compounds from Leaves of Carya cathayensis on AngiotensinⅡ-induced Proliferation and Migration in Rat Vascular Smooth Muscle Cells

目的：研究山核桃叶总黄酮及其4种单体化合物球松素查尔酮、松属素、白杨素、汉黄芩素对血管紧张素Ⅱ（angiotensin Ⅱ，AngⅡ）诱导的血管平滑肌细胞（vascular smooth muscle cells，VSMC）增殖和迁移的作用。方法：组织贴块法培养大鼠胸主动脉VSMC。调细胞密度2．0×10^4／mL，血清饥饿处理后用100nmol·L^-1 Ang Ⅱ诱导增殖12h。调细胞密度6．0×10^4／mL，用100nmol·L^-1 Ang Ⅱ诱导迁移12h。均给予12．5～62．5mg·L～总黄酮，10～50mg·L^-1球松素查尔酮、松属素、白杨素、汉黄芩素作用12h。用细胞计数法检测VSMC增殖，伤口愈合实验检测VSMC迁移。结果：12．5mg·L～总黄酮可剂量依赖性地显著抑制AngII诱导的VSMC增殖，50mg·L。球松素查尔酮、白杨素和汉黄芩素对AngⅡ诱导的VSMC增殖有微弱抑制。12．5mg·L^-1总黄酮，10mg·L^-1球松素查尔酮、松属素、白杨素、汉黄芩素均能剂量依赖性地显著抑制AngⅡ诱导的VSMC迁移。其中总黄酮、白杨素、汉黄芩素的效果优于松属素和球松素查尔酮。结论：山核桃叶总黄酮能有效抑制AngII诱导的VSMC增殖和迁移；白杨素、汉黄芩素、松属素和球松素查尔酮对AngⅡ诱导的VSMC迁移的抑制作用强于对其诱导的VSMC增殖的抑制作用。

Objective： To investigate the inhibitory effects of total flavonoids from the leaves of Carya cathayensis and their four monomer compounds, namely pinostrobin chalcone, pinocembrin, chrysin and wogonin on angiotensin Ⅱ（ Ang Ⅱ ） -induced proliferation and migration in rat vascular smooth muscle cells （VSMC）. Method： VSMC were primary cultured by tissue sticking method. VSMC proliferation was induced by 100 nmol .L^-1 Ang Ⅲ for 12 h after treatment of serum starvation at the initial density of 2.0 × 10^4/mL. VSMC migration was induced by 100 nmol .L^-1 Ang Ⅱ for 12 h at the initial density of 6.0 × 10^4/mL. Both of them were treated with 12.5-62.5 mg.L^-1＇ total flavonoids, 10-50 mg .L^-1 pinostrobin chalcone, pinocembrin, chrysin and wogonin for 12 h. Cell proliferation was measured by direct counting the cells and cell migration was determined using the wound healing assay, respectively. Result： 12.5 mg.L^-1 total flavonoids significantly inhibited Ang 11-induced VSMC proliferation in a does dependent manner while 50 mg. L^-1 pinostrobin ehaleone, chrysin and wogonin slightly inhibited Angll-induced VSMC proliferation. 12.5 mg .L^-1 total flavonoids, 10 mg .L^-1 pinostrobin chalcone, pinoeembrin, ehrysin and wogonin significantly inhibited Ang Ⅱ-induced VSMC migration in a does dependent manner. Among them, the total flavonoids, chrysin and wogonin showed a higher inhibitory effect compared with those of pinocembrin and pinostrobin chalcone. Conclusion： The total flavonoids fi＇om the leaves of C. cathayensis can effectively inhibit Ang Ⅱ-induced VSMC proliferation and migration, while their four monomer compounds present better suppressive activities on Ang Ⅱ -induced migration than those of proliferation in VSMC.