西洋参茎叶体外抑制大肠杆菌增殖及抗炎活性的研究

Inhibitory effect of stems and leaves of Panax quinquefolium on proliferation and anti-inflammatory activity of Escherichia coli in vitro

为探讨西洋参茎叶不同提取部位对小鼠单核巨噬细胞RAW264.7抑菌效果及脂多糖(LPS)诱导的抗炎活性的影响,本试验用106 CFU/mL大肠杆菌菌液侵染RAW264.7细胞,分别以西洋参茎叶水提取和醇提取部位0.125、0.25、0.5 mg/mL作用细胞,CCK-8法检测不同提取物浓度对RAW264.7细胞增殖活性的影响,Hoechst染色法观察细胞被大肠杆菌侵染后的凋亡情况。LPS诱导细胞炎症反应,ELISA法检测细胞炎性因子IL-6、IL-1β、TNF-α的释放情况;实时荧光定量PCR法测定细胞中细胞炎性因子IL-6、IL-1β、TNF-αmRNA表达。结果表明:西洋参茎叶水提取部位和醇提取部位能抑制大肠杆菌诱导的细胞凋亡;西洋参茎叶水提取部位提取物浓度为0.25 mg/mL时,细胞释放TNF-α、IL-1β和IL-6细胞炎症因子浓度分别降低了50.20%、22.21%、43.41%,对其mRNA基因表达量分别降低了12.31%、27.07%、29.01%;西洋参茎叶醇提取部位提取物浓度为0.25 mg/mL时,细胞释放TNF-α、IL-1β和IL-6细胞炎症因子浓度分别降低了59.70%、30.24%、52.38%,对其mRNA基因表达量分别降低了21.54%、29.84%、31.79%,且醇提取部位较水提取部位的抑制效果好。西洋参茎叶水提取部位和醇提取部位对大肠杆菌有抑制作用,能增强细胞抗炎活性,为西洋参茎叶作为中草药饲料添加剂进一步开发利用提供了理论依据。

To investigate the effects of different extraction parts of stem and leaf of Panax quinquefolium on the antibacterial effect of mouse monocyte macrophage RAW264.7 and the anti-inflammatory activity induced by lipopolysaccharide(LPS).RAW264.7 cells were infected with 106 CFU/mL E.coli bacterial solution.The cells were treated with 0.125,0.25 mg/mL and 0.5 mg/mL of water and alcohol extracts from stems and leaves of Panax quinquefolium,respectively.CCK-8 method was used to detect the effect of different extract concentrations on the proliferation activity of RAW264.7 cells.Hoechst staining was used to observe the apoptosis of cells after infection with E.coli.LPS induced cellular inflammatory response,and the release of IL-6,IL-1βand TNF-αwas detected by ELISA.The mRNA expressions of IL-6,IL-1βand TNF-αin cells were determined by real-time fluorescent quantitative PCR.The results showed as follows:Water extract and alcohol extract from stems and leaves of Panax quinquefolium could inhibit the apoptosis induced by Escherichia coli.When the concentration of water extract from stems and leaves of Panax quinquefolium was 0.25 mg/mL,the concentration of TNF-α,IL-1βand IL-6 released by cells decreased by 50.20%,22.21%and 43.41%,respectively,and the mRNA gene expression decreased by 12.31%,27.07%and 29.01%,respectively.When the ethanol extract concentration was 0.25 mg/mL,the concentration of TNF-α,IL-1βand IL-6 released by cells decreased by 59.70%,30.24%and 52.38%,respectively,and the mRNA gene expression decreased by 21.54%,29.84%and 31.79%,respectively.The inhibitory effect of ethanol extract was better than that of water extract.The water extraction and alcohol extraction parts of stems and leaves of Panax quinquefolium have inhibitory effects on E.coli,and could enhance the anti-inflammatory activity of cells,which provides a theoretical basis for the further development and utilization of stems and leaves of Panax quinquefolium as feed additives of Chinese herbal medicine.