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目的观察氧化低密度脂蛋白(ox LDL)诱导血管内皮细胞(VEC)损伤条件培养基及气血并治方有效组分D(主要为芍药苷)和E(主要为总黄酮)配伍(重量比1∶2)是否对血管平滑肌细胞(VSMC)MAPK、PKC活性和胞内Ca2+荧光强度有影响。方法将VSMC正常培养液、ox LDL诱导VEC损伤的条件培养基、辛伐他汀加ox LDL诱导VEC损伤的条件培养基、有效组分D和E优化配伍加ox LDL诱导VEC损伤的条件培养基等分别作用于VSMC,采用β放射活性法测定丝裂素活化蛋白激酶(mitogen activatedproteinkinase,MAPK)及蛋白激酶C(proteinkinaseC,PKC)活性,流式细胞仪检测VSMC内的Ca2+荧光强度。结果VEC损伤条件培养基作用于VSMC后,与正常培养VSMC相比,细胞MAPK、PKC活性明显增加(P<0.01),细胞内Ca2+荧光强度增加;有效组分D和E配伍作用于VEC损伤条件培养基培养的VSMC后,与模型组相比,MAPK及PKC活性明显减少(P<0.01)、细胞内Ca2+荧光强度降低。结论气血并治方中有效组分D和E优化配伍可以拮抗MAPK、PKC活性和细胞内Ca2+荧光强度的增加,气血并治方防治动脉粥样硬化(AS)的作用可能与此有关。
Objective To observe the oxidative low-density lipoprotein (ox-LDL)-induced vascular endothelial cell (VEC) injury conditional medium and the effective components D (mainly paeoniflorin) and E (mainly total flavonoids) in the combination of Qixue Bingfang prescription (weight ratio). 1 : 2) Whether it affects the MAPK, PKC activity and intracellular Ca 2+ fluorescence intensity of vascular smooth muscle cells (VSMCs). Methods VSMC normal culture medium, conditioned medium for VEC injury induced by ox LDL, conditioned medium for VEC damage induced by simvastatin plus ox LDL, and optimized combination of effective components D and E plus ox LDL for conditioned medium for VEC injury The activity of mitogen-activated protein kinase (MAPK) and protein kinase C (PKC) was measured by beta-irradiation method. The Ca2+ fluorescence intensity of VSMC was detected by flow cytometry. Results Compared with normal cultured VSMCs, VEC injury conditioned medium significantly increased the activity of MAPK and PKC (P<0.01), and increased the intracellular Ca2+ fluorescence intensity. The effective components D and E were compatible with VEC damage conditions. After VSMC cultured in culture medium, compared with the model group, the activity of MAPK and PKC was significantly decreased (P<0.01), and the intracellular Ca2+ fluorescence intensity was decreased. Conclusion The optimized combination of effective components D and E in Qixue Bingzhi Formula can antagonize the increase of MAPK, PKC activity and intracellular Ca2+ fluorescence intensity. The effect of Qixuebingzhi Prescription on prevention and treatment of atherosclerosis (AS) may be related to this.