【作者】 刘中勇； 李林； 方家； 陈宁南； 邓鹏； 方险峰； 邹国辉；

【机构】 江西中医药大学附属医院心血管病科； 江西省人民医院；

【摘要】 目的:分离鉴定人脐静脉内皮原代细胞,并观察补肾活血方剂对原代细胞的NO分泌促进作用。方法:采用Ⅰ型胶原酶消化分离人脐静脉内皮细胞,培养传代。应用相差显微镜观察细胞形态。免疫荧光染色证明新分离细胞表达Ⅷ相关抗原。制备兔补肾活血含药血清,分为高中低及空白(含新生胎牛血清)对照组(不含药兔血清)。分别作用原代细胞。Western blot检测eNOS蛋白表达,NO检测试剂盒检测细胞上清液NO含量。结果:细胞形态学和免疫荧光结果显示,成功分离人脐静脉内皮细胞。补肾活血方剂效提高eNOS抑制剂L-LAME处理的细胞表达eNOS及NO分泌。结论:补肾活血方能有效提高内皮细胞NO分泌。是一种潜在提高血管通透性的潜在处方,具体机制有待进一步探讨。更多还原

【Abstract】 Objective:separation of human umbilical vein endothelial, and observe the effect of BuShengHuoXue(BSHX) herbal formal promoting NO secretion.Methods:using collagenase type I digestion separation human umbilical vein endothelial cells,training batches,using phase contrast microscope observation the cell morphology.Immunofluorescence staining proved that the new separated cells express VI related antigens. Preparation of the drug-containing serum from rabbit.Set blank (including new fetal bovine serum) and control group(excluding rabbit serum).Primitive cells function respectively.Western blot used to detecting the eNOS protein expression.The kits with NO measurement cell supernatant NO content.Results:according to the cell morphology and immunofluorescence,we had successful separated human umbilical vein endothelial cells.BSHX prescription can effectively improve the eNOS inhibitor treated cells secreting NO.We should do more work to find the machine.更多还原