清毒活血化痰复方抑制脐静脉内皮细胞株ECV304氧糖剥夺/复糖复氧模型中NF-κB和STAT3的表达  

Qingdu Huoxue Huatan Compound inhibits the expression of nf-kappa B and STAT3 in umbilical vein endothelial cell line ECV304 OGD/R model

作　　者：骆殊[1] 邵佳[1] 吴颢昕[1] 陈刚[1] 刘舟[1] 

机构地区：[1]南京中医药大学,南京210046

出　　处：《北京中医药大学学报》2015年第2期120-125,共6页Journal of Beijing University of Traditional Chinese Medicine

基　　金：江苏省自然科学基金资助项目(No.BK2007243);江苏省高校自然科学研究项目(No.13KJD360001);南京中医药大学青年自然科学基金项目资助(No.12XZR07)

摘　　要：目的探讨清毒活血化痰复方对脐静脉内皮细胞氧糖剥夺/复糖复氧(OGD/R)损伤的保护作用及其作用机制。方法 SD大鼠连续5 d每天灌胃清毒活血化痰复方(12 g/kg),正常对照组给予等量生理盐水,制备药物血清和正常血清。OGD/R损伤脐静脉内皮细胞株ECV304,复氧复糖过程中添加清毒活血化痰复方药物血清,并以正常血清为对照。正常对照组、缺氧缺糖后复氧复糖组(模型组)、清毒活血化痰复方药物血清组(中药组)分别于复氧复糖后4、24、48、72 h提取基因和蛋白用于检测。以MTT法检测细胞存活率,用RT-PCR检测NF-κB和STAT3的mRNA水平,Western blot检测NF-κB和STAT3的蛋白表达。结果清毒活血化痰复方药物血清作用24 h即显现出明显的促ECV304增殖的作用,于48 h时作用最明显。在各个时间段对NF-κB和STAT3的mRNA和蛋白表达均有抑制作用。结论清毒活血化痰复方药物血清可以有效改善OGD/R对脐静脉内皮细胞的损伤,促进细胞增殖,其作用机制可能与抑制NF-κB和STAT3的表达相关。Objective To study the protective effects of Qingdu Huoxue Huatan（ Toxin-removing bloodactivating and phlegm-resolving） compound against OGD / R damage to umbilical vein endothelial cell and its underlying mechanism. Methods SD rats were intragastrically given Qingdu Huoxue Huatan compound（ 12 g / kg） while the control group received normal saline of the same dosage. Sera of rats in two groups were extracted afterwards. Drug serum of Qindu Huoxue Huatan compound was added to the OGD / R-damaged umbilical vein endothelial cell line ECV304 in the process of regainning oxygen and sugar,and cell survival rate was evaluated by using MTT and mRNA; protein levels of NF-κB and STAT3 were measured by using rt-PCR and western blotting. Results After 24 h,Qindu Huoxue Huatan compound drug serum promoted ECV304 proliferation,and the promotion reached the peak at 48 h.Inhibition of mRNA and protein of NF-κB and STAT3 was observed at each time period.Conclusion Qindu Huoxue Huatan compound drug serum could effectively improve OGD / R damage to umbilical vein endothelial cell line ECV304,and promote cell＇s proliferation. Its mechanism may be related to the inhibited expressions of nf-kappa B and STAT3.

关 键 词：清毒活血化痰复方 脐静脉内皮细胞 氧糖剥夺/复糖复氧 NF-ΚB STAT3 大鼠 

分 类 号：R285.5[医药卫生—中药学]