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目的:研究中药地骨皮醇提液对细菌脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVEC)炎症的保护作用,为研究糖尿病新药提供理论基础。方法:将HUVEC细胞分为正常对照组、LPS对照组和药物预处理+LPS作用组,作用24h后。倒置显微镜观察细胞的形态学变化。采用ELISA法测定细胞上清中所分泌的ICAM-1量。并通过RT-PCR法检测细胞因子ICAM-1的表达。结果:LPS对照组细胞出现了明显的炎症感染表现,而用药物处理组细胞的状态出现了明显的好转。ELISA结果显示低浓度的地骨皮提取液相对单独LPS作用组并不能降低上清ICAM-1的分泌量,但中高浓度的地骨皮提取液可以降低ICAM-1的分泌量,且具有统计学意义。RT-PCR表明LPS能显著激活ICAM-1mRNA的表达,而当给予了地骨皮提取液后,除低剂量组外,中高剂量组能较明显降低ICAM-1mRNA的表达。结论:地骨皮醇提液能够对细菌脂多糖(LPS)诱导的人脐静脉内皮细胞的损害有保护作用,这可能成为地骨皮防治糖尿病血管病变的一个机制。
OBJECTIVE: To study the protective effect of extract of Digupi of Chinese medicine on inflammation induced by bacterial lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVEC), and to provide a theoretical basis for the study of new drugs for diabetes. Methods: HUVECs were divided into normal control group, LPS control group and drug pretreatment + LPS group. Inverted microscope was used to observe the morphological changes of cells. The amount of ICAM-1 secreted in the cell supernatant was measured by ELISA. The expression of cytokine ICAM-1 was detected by RT-PCR. Results: The cells in LPS control group showed obvious inflammatory infection, while the cells treated with drug showed obvious improvement. ELISA results showed that low concentration of Digupi extract relative LPS alone group can not reduce the secretion of ICAM-1 supernatant, but the high concentration of Digupi extract can reduce the secretion of ICAM-1, and with statistical significance. RT-PCR showed that LPS could significantly activate the expression of ICAM-1 mRNA. However, when the extract from Digupi skin was treated with LPS, the expression of ICAM-1 mRNA was significantly lower than that of the low-dose group. CONCLUSION: The extract of Digupi can protect LPS-induced injury of human umbilical vein endothelial cells, which may be a mechanism of prevention and treatment of diabetes mellitus.