摘要
目的探讨麝香保心丸对过氧化氢诱导人脐静脉内皮细胞增殖凋亡及炎症因子表达的影响。方法体外胶原酶消化法培养人脐静脉内皮细胞,5-溴脱氧尿嘧啶核苷渗入法测定细胞增殖活性,dUTP缺口末端标记法测定内皮细胞的凋亡,逆转录-聚合酶链反应法测定炎症因子单核细胞趋化蛋白1、白细胞介素6、核因子-κB-P65 mRNA表达水平,W estern b lotting测定内皮细胞核因子-κB-P65的蛋白水平。结果 (1)与对照组相比,过氧化氢组内皮细胞活性明显降低(P<0.05),麝香保心丸药物血清呈浓度依赖性抑制过氧化氢诱导内皮细胞损伤(P<0.05),与对照血清组相比,药物血清1 g组内皮细胞活性显著升高(P<0.05);(2)过氧化氢组内皮细胞大量凋亡,1 g麝香保心丸血清组只有少量细胞凋亡;(3)与对照组相比,过氧化氢组单核细胞趋化蛋白1、白细胞介素6、核因子-κB-P65 mRNA表达水平明显增高(P<0.05),麝香保心丸药物血清组呈浓度依赖性抑制上述指标的表达,1 g麝香保心丸血清组有明显降低,而对照血清组单核细胞趋化蛋白1、白细胞介素6、核因子-κB-P65 mRNA表达无明显差别(P>0.05);(4)与对照组相比,过氧化氢组核因子-κB-P65蛋白表达水平明显增高,麝香保心丸(血清)呈浓度依赖性抑制其的表达,1 g麝香保心丸血清组核因子-κB-P65表达水平明显降低(P<0.05)。结论麝香保心丸保护血管内皮细胞可能与抑制过氧化氢诱导人脐静脉内皮细胞炎症因子有关。
Aim To investigate the effect of Shexiang Baoxin Wan on cell proliferation,apoptosis and expression of inflammatory factors in human umbilicial vein endothelial cells（HUVEC） induced with hydrogen peroxide（H2O2）. Methods Collagenase I was used to digest and isolate HUVEC and then cultured.Bromodeoxyuridine（BrdU） enzyme-linked immunosorbent assay（ELISA） was used to assay proliferation of HUVEC in vitro.Cell apoptosis was determined by terminal dUTP nick end labling（TUNEL）.Expression of monocyte chemotactic protein-1（MCP-1）,interleukin-6（IL-6） and nuclear factor-kappa B（NF-κB）-P65 mRNA was detected by reverse transcription polymerase chain reaction（RT-PCR）.Western-blotting was performed to detect the expression of NF-κB-p65. Results（1）Compared with control,cell growth and proliferation was significantly lower in H2O2 group.The serum of Shexiang Baoxin Wan could inhibit the HUVEC proliferation declined by H2O2 in concentration-dependent manner.Apoptosis was higher in H2O2 groups than that in control group and reserved to normal in 1 g Shexiang Baoxin Wan.（2）Compared with control,the level of MCP-1,IL-6 and NF-κB mRNA in H2O2 group was greatly increased.Shexiang Baoxin Wan could inhibit the expression of these inflammatory factors induced by H2O2 in concentration-dependent manner.The serum of 1 g Shexiang Baoxin Wan remarkly decreased the level of MCP-1,IL-6 and NF-κB-P65 mRNA.（3） Compared to control,the level of NF-κB-P65 was largely increased in H2O2 group,Shexiang Baoxin Wan could inhibit the expression of NF-κB-P65 in concentration-dependent manner.The serum of 1 g Shexiang Baoxin Wan greatly decreased the level of NF-κB-P65. Conclusion The role of Shexiang Baoxin Wan in protecting endothelial cell may be related to inhibiting the inflammatory factors induced by H2O2.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2011年第10期813-818,共6页
Chinese Journal of Arteriosclerosis
关键词
麝香保心丸
内皮细胞
炎症因子
Shexiang Baoxin Wan
Endothelial Cells
Inflammatory Factors
