模拟手法压力刺激对颈性眩晕血瘀证患者血清损伤后内皮细胞的影响

Effect of Stress Stimulation of Manipulation on Endothelial Cells with Serum Induced Injury in Patients with Blood Stasis Syndrome of Cervical Vertigo

目的观察模拟手法压力刺激对颈性眩晕血瘀证患者血清损伤的人脐静脉内皮细胞的影响及机制。方法将培养的颈性眩晕血瘀证患者血清损伤的人脐静脉内皮细胞(HUVEC-C)分为血瘀证组(DMEM+10%患者血清+0 mm Hg)、低压组(DMEM+10%患者血清+90 mm Hg)和高压组(DMEM+10%患者血清+180 mm Hg),健康对照组(DMEM培养液细胞+10%健康人血清+0 mm Hg),于相应压力干预实验后,倒置显微镜下观察其形态学变化,MTT法检测其活性,硝酸还原酶法检测一氧化氮(NO)浓度,放射免疫分析技术测定内皮素(ET)。结果经过低压力刺激干预后细胞形态有一定变化,细胞活性有所提高,增加NO释放、抑制细胞ET释放,而高压力刺激则无显著影响。结论颈性眩晕血瘀证患者血清可以损伤血管内皮细胞,适当的压力刺激能增强细胞的活性和维持细胞的形态结构,压力刺激调节血管内皮细胞血管舒缩活性物质ET、NO合成释放,这可能是手法发挥活血化瘀功效的细胞生物学机制之一。

Objective： To study the effect and mechanism of stress stimulation of manipulation on endothelial cells with serum induced injury in patients with blood stasis syndrome of cervical vertigo. Methods： The cultivat- ed HUVEC-C of injury induced by the serum of the patient of blood stasis syndrome （BSS） were divided into blood stasis syndrome group （DMEM＋10% serum of the patient＋0 mmHg）,low pressure group （DMEM＋10% serum of the patient＋90 mmHg）,high pressure group （DMEM＋10% serum of the patient ＋180 mmHg） and healthy control group （DMEM cell of nutrient solution＋10% healthy human serum＋0 mmHg）. After corresponding stress intervenes in the experiment, the change of its configuration was put upside down under the microscope and observed,and its activity was tested by MTT assay,nitric reductase assay to measure NO concentration and ra- dioimmunoassay （RIA） to measure ET. Results： After intervention of low stress stimulation, the cell configuration had some changes. Cell activity increased. It could increase NO release and inhibit release of ET. However,there was no obvious effect under the high stress stimulation. Conclusion ： The serum of the patient of blood stasis syn- drome （BSS） can injure the vascular endothelial cell. Appropriate stress stimulation can increase the cell activity and maintain the cell configuration. Stress stimulation can regulate the synthetic release of ET and NO of vascular active substances in vascular endothelial ceil,which may be one of mechanisms of cell biology through technique effects of activating blood circulation to dissipate blood stasis.