槲皮素对HaCat细胞增殖及β-防御素和谷胱甘肽过氧化物酶表达的影响

Effects of quercetin on the proliferation of HaCat cells and the expression of β-defensin and glutathione peroxidase

目的研究槲皮素(quercetin)对人永生化上皮细胞(HaCat细胞)增殖及其非特异性免疫分子表达的影响。方法利用MTT法检测不同浓度槲皮素溶液作用对HaCat细胞增殖的影响,利用ELISA法检测在不同浓度槲皮素干预下的HaCat细胞培养上清液中β-防御素(β-DF)及谷胱甘肽过氧化物酶(GPX)的含量。结果 100μmol/L槲皮素对HaCat细胞的增殖有抑制作用,50μmol/L以下的槲皮素对HaCat细胞的增殖无明显影响。HaCat细胞经过不同浓度的槲皮素处理48、72h后,其细胞上清液中谷胱甘肽过氧化物酶的浓度与对照组比较差异有统计学意义(P<0.05),但β-防御素的表达在50.00μmol/L和25.00μmol/L槲皮素处理组上调。结论槲皮素具有增强细胞非特异免疫功能作用,其机制可能通过上调β-防御素的表达来实现。

Objective To study the effect of quercetin on the proliferation of HaCat cells and the expression of nonspecific immunity molecules（glutathione peroxidase andβ-defensin）.Methods The proliferation of HaCat cells after treatment with different concentrations of quercetin（100.00,50.00,25.00,12.50,6.25,and 0.00μmol/L）was detected using an MTT assay.The concentrations ofβ-defensin and glutathione peroxidase in HaCat cell culture supernatants treated with different concentrations of quercetin were detected with ELISA. Methods The proliferation of HaCat cells after treatment with different concentrations of quercetin（100.00,50.00,25.00,12.50,6.25,and 0.00μmol/L）was detected using an MTT assay.The concentrations ofβ-defensin and glutathione peroxidase in HaCat cell culture supernatants treated with different concentrations of quercetin were detected with ELISA. Results The proliferation of HaCat cells was significantly inhibited by 100μmol/L quercetin（compared to treatment with 0.00μmol/L quercetin;the t values was 9.84 12 hafter treatment with 100.00μmol/L quercetin,13.33 24 hafter treatment,11.34 48 hafter treatment,11.84 72 hafter treatment,and 14.88 96 hafter treatment,P〉0.05 for all）.However,a concentration of quercetin less than 50μmol/L had no effect on HaCat cell proliferation（compared to treatment with 0.00μmol/L quercetin;the Fvalues were 2.02 12 hafter treatment with 50.00,25.00,12.50,or 6.25μmol/L quercetin,2.46 24 hafter treatment,0.59 48 hafter treatment,2.72 72 hafter treatment,and 2.75 96 hafter treatment（P〈0.05 for all）.The concentration of glutathione peroxidase（GPX）in the cell culture supernatant 48 hand 72 hafter treatment with quercetin decreased markedly compared to that in the control group（the Fvalues were 24.53 and 5.78,respectively,P〈0.05 for both）.Cells treated with 50.00,25.00,12.50,or 6.25μmol/L quercetin for 48 hhad a significantly lower concentration of GPX in the supernatant（14.29±0.58 pg/mL,14.11±0.26 pg/mL,16.21±0.76 pg/mL and 16.60±0.52 pg/mL）compared to that in the control group（18.40 ± 1.15 pg/mL）.Cells treated with 25.00μmol/L or 12.50μmol/L quercetin for 72 hhad a significantly lower concentration of GPX in the supernatant（14.9 ± 1.07 pg/mL and 15.19 ± 0.69pg/mL）compared to that in the control group（17.05±1.25pg/mL,the q values were 4.46and 3.86,P〈0.05for both）.The concentration ofβ-defensin（β-DF）in the supernatant of cells treated with different concentrations of quercetin for 48hdid not differ significantly（F=2.56.P〉0.05）,but concentration ofβ-DF in the supernatant of cells treated with 50.00μmol/L or 25.00μmol/L quercetin for 24hand 72hwas 435.87±55.94pg/mL and 406.61±20.56pg/mL and 480.93±31.00pg/mL and 475.22±42.93pg/mL,respectively.The concentration ofβ-DF increased markedly compared to that in the control group（355.84±18.81pg/mL and 374.29±23.87pg/mL;the t values were 2.71and3.64and 5.45and 4.18,respectively,P〈0.05for all）.Conclusion Quercetin may enhance the expression ofβ-DF,thus enhancing nonspecific immunity.