机构地区:[1]510632广州,暨南大学附属第一医院普外科 香港圣母医院外科部 [2]510632广州,暨南大学附属第一医院普外科 510632广州,暨南大学附属第一医院体部伽玛刀中心 [3]暨南大学医学院组织胚胎学教研室
出 处:《中华实验外科杂志》2014年第2期312-315,共4页Chinese Journal of Experimental Surgery
基 金:国家基础研究973计划资助项目(2010CB833603);国家自然科学基金资助项目(30772131);暨南大学第一临床医学院科研培育基金资助项目(511005004);暨南大学第一临床医学院开放基金资助项目(511005026)
摘 要:目的 观察纳米金(AuNP)对HepG2细胞血管内皮生长因子(VEGF)表达和分泌的影响.方法 实验分为不同浓度AuNP(5、10、20 μg/L)处理组和对照组,HepG2细胞常规消化种板,培养24h后:AuNP处理组分别加入浓度为5、10、20 μg/L的AuNP溶液1 ml;对照组加入等量无血清培养液,继续培养12h后:采用Western blot法检测HepG2细胞内VEGF的表达水平;酶联免疫吸附试验(ELISA)检测HepG2细胞VEGF的分泌水平;噻唑蓝(MTT)比色法检测HepG2细胞增殖率.改变HepG2细胞的培养条件(37℃/4℃),观察AuNP对HepG2细胞VEGF分泌的影响.建立肝癌腹水瘤模型进一步观察AuNP对模型小鼠腹水VEGF水平及腹水量的影响.结果 体外实验:各组HepG2细胞分泌的VEGF浓度分别为:对照组(351.64±7.89)ng/L;5μg/L AuNP组(285.62±3.45) ng/L;10 μg/L AuNP组(121.72±3.10) ng/L;20 μg/L AuNP组(9.83±2.28) ng/L,P<0.05.各AuNP处理组(5、10、20 μg/L) HepG2细胞增殖率分别为:(98.98±7.57)%、(91.09±11.46)%、(92.13±5.65)%,各组AuNP对HepG2细胞增殖影响差异均无统计学意义(P>0.05).体内实验:腹水瘤模型小鼠腹水VEGF水平及腹水体积分别为:对照组(62.95±11.93) ng/L、(13.22±1.03) ml;AuNP处理组(27.12 ±8.58) ng/L、(5.21 ±0.62) ml,P<0.01.结论 AuNP可以明显抑制HepG2细胞VEGF的表达和分泌,体内实验证实AuNP能够降低腹水瘤模型小鼠腹水VEGF水平,明显减少腹水量.Objective To observe the inhibitory effect of gold nanoparticles on the expression and secretion of vascular endothelial growth factor (VEGF) by HepG2 cells.Methods HepG2 cells were seeded in 96-well plates and treated with different concentrations (5,10,and 20 μg/L) of gold nanoparticles for 12 h.The concentration of VEGF protein expressed and secreted by HepG2 cells was determined by using Western blotting and enzyme linked immunosorbent assay (ELISA) separately.The proliferation rate of HepG2 cells was assessed by using methyl thiazol tetrazolium (MTT) assay.For mouse ascites model experiments,H22 cells were intraperitoneally injected into BALB/c mice.After 7 days,gold nanoparticles (1 mg/kg) was administered through intraperitoneal injection every 2 days over a period of 14 days.Mice were then sacrificed and ascites fluids collected.The fluid accumulation in the peritoneal carvity,as well as the concentration of VEGF of mouse ascites was determined.Results The VEGF concentration of 4 groups (control,5,10 and 20 μg/L gold nanoparticles) was (351.64 ±7.89),(285.62 ± 3.45),(121.72 ±3.10) and (9.83 ±2.28) ng/L respectively,P 〈0.05.The proliferation rate of HepG2 cells in three gold nanoparticles group (5,10,and 20 μg/L) was (98.98 ±7.57)%,(90.95 ± 11.62)% and (86.38 ±4.23)% respectively,P 〉 0.05.The ascites accumulation and VEGF concentration in control and gold nanoparticles groups were (13.22 ± 1.03) ml and (62.95 ± 11.93) ng/L,and (5.21 ± 0.62) ml and (27.12 ± 8.58) ng/L respectively,P 〈 0.01.Conclusion Gold nanoparticles can inhibit the expression and secretion of VEGF and cause lesser ascites accumulation of mouse ascites model.
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