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目的:探讨清毒活血化痰复方颗粒对内皮细胞氧糖剥夺-复糖复氧(OGD/R)模型ERK1/2、STAT3蛋白磷酸化和ICAM-1表达的调控作用。方法:SD大鼠连续5 d灌胃清毒活血化痰复方颗粒(12 g/kg)或等体积生理盐水,制备药物血清和空白血清。脐静脉内皮细胞株ECV304分为正常对照组、氧糖剥夺/复糖复氧模型组、清毒活血化痰复方颗粒药物血清组、清毒活血化痰复方颗粒药物血清+U0126(ERK1/2磷酸化特异性阻断剂)组,分别于复氧复糖后4、24、48、72 h以Western blot检测p-ERK1/2、p-STAT3及ICAM-1蛋白的表达,RT-PCR检测ICAM-1 mRNA的表达。结果:作用24 h清毒活血化痰复方颗粒药物血清可以明显促进ERK1/2的磷酸化,并抑制STAT3磷酸化及ICAM-1的表达,但U0126可以部分阻断清毒活血化痰复方颗粒药物血清的以上作用。结论:清毒活血化痰复方颗粒药物血清可通过ERK1/2/STAT3通路调控OGD/R损伤脐静脉内皮细胞株ECV304 ICAM-1的表达。
Objective: To investigate the regulation of the expression of phosphorylation of ERK1 / 2, STAT3 protein and ICAM-1 in the model of OGD / R with endothelial cell oxygen-deprivation and replenishing phlegm compound granules. Methods: SD rats were treated with Qingxuhuoxue Huatan Compound Granule (12 g / kg) or normal saline for 5 days continuously to prepare drug serum and blank serum. Human umbilical vein endothelial cell line ECV304 was divided into normal control group, Oxygen-glucose deprivation / rewarming model group, Qingduhuoxue Huatan Compound Granule drug serum group, Qingdu Huoxue Huatan Compound Granule drug serum + U0126 (ERK1 / 2 phosphate Specific blockers). The expressions of p-ERK1 / 2, p-STAT3 and ICAM-1 protein were detected by Western blot at 4, 24, 48 and 72 h after reoxygenation treatment respectively. -1 mRNA expression. Results: The effect of 24 h serum of drug-promoting and blood-activating and resolving phlegm compound drugs could obviously promote the phosphorylation of ERK1 / 2 and the STAT3 phosphorylation and the expression of ICAM-1, but U0126 could partially block the drug Serum above effect. Conclusion: The drug serum of QDDHR can regulate the expression of ICAM-1 in umbilical vein endothelial cells (ECV304) induced by OGD / R through ERK1 / 2 / STAT3 pathway.