CAJ | 학술논문

目的：探索原花青素B1的抗炎活性及其抗炎机制。方法以人急性单核细胞白血病细胞系（THP-1）为模型，设立不含脂多糖（LPS）的磷酸盐缓冲溶液（PBS）处理THP-1细胞为对照组，检测不同浓度原花青素B1（50、100、150、200 mg/L）对THP-1细胞活力的影响。分别应用1 mg/L的LPS（LPS组）、1 mg/L的LPS联合原花青素B150 mg/L（处理A组）、1 mg/L的LPS联合原花青素B1100 mg/L（处理B组）预处理THP-1细胞，收集各组THP-1细胞上清液，应用双抗体夹心亲和素-生物素-过氧化物酶复合物-酶联免疫吸附法测定各组THP-1细胞中TNF-α的释放水平。应用实时定量PCR法测定各组THP-1细胞MyD88 mRNA和髓样分化蛋白2（MD-2） mRNA的表达情况。结果不同浓度原花青素的THP-1细胞活力与对照组比较，差异有统计学意义（F=31.35，P<0.001），进一步比较发现，50、100 mg/L原花青素的THP-1细胞活力与对照组比较差异均无统计学意义（P均＞0.05），而150、200 mg/L原花青素的THP-1细胞活力均明显低于对照组（P均＜0.05）。处理A、B组的TNF-α水平较LPS组均显著减低（P均＜0.05），且处理B较处理A组下降更为明显（P＜0.05）。对照组、LPS组及处理A组间MyD88 mRNA和MD-2 mRNA的表达差异均有统计学意义（F=62.500、46.700，P均＜0.001），且LPS组的MyD88 mRNA和MD-2 mRNA表达均较对照组及处理A组明显升高（P均＜0.05），处理A组的MyD88 mRNA和MD-2 mRNA表达高于对照组（P均＜0.05）。结论原花青素B1可明显抑制LPS介导的炎症反应，负性调节Toll样受体-MyD88信号通路，可能是其抑制炎症反应的潜在机制。
목적：탐색원화청소B1적항염활성급기항염궤제。방법이인급성단핵세포백혈병세포계（THP-1）위모형，설립불함지다당（LPS）적린산염완충용액（PBS）처리THP-1세포위대조조，검측불동농도원화청소B1（50、100、150、200 mg/L）대THP-1세포활력적영향。분별응용1 mg/L적LPS（LPS조）、1 mg/L적LPS연합원화청소B150 mg/L（처리A조）、1 mg/L적LPS연합원화청소B1100 mg/L（처리B조）예처리THP-1세포，수집각조THP-1세포상청액，응용쌍항체협심친화소-생물소-과양화물매복합물-매련면역흡부법측정각조THP-1세포중TNF-α적석방수평。응용실시정량PCR법측정각조THP-1세포MyD88 mRNA화수양분화단백2（MD-2） mRNA적표체정황。결과불동농도원화청소적THP-1세포활력여대조조비교，차이유통계학의의（F=31.35，P<0.001），진일보비교발현，50、100 mg/L원화청소적THP-1세포활력여대조조비교차이균무통계학의의（P균＞0.05），이150、200 mg/L원화청소적THP-1세포활력균명현저우대조조（P균＜0.05）。처리A、B조적TNF-α수평교LPS조균현저감저（P균＜0.05），차처리B교처리A조하강경위명현（P＜0.05）。대조조、LPS조급처리A조간MyD88 mRNA화MD-2 mRNA적표체차이균유통계학의의（F=62.500、46.700，P균＜0.001），차LPS조적MyD88 mRNA화MD-2 mRNA표체균교대조조급처리A조명현승고（P균＜0.05），처리A조적MyD88 mRNA화MD-2 mRNA표체고우대조조（P균＜0.05）。결론원화청소B1가명현억제LPS개도적염증반응，부성조절Toll양수체-MyD88신호통로，가능시기억제염증반응적잠재궤제。
Objective To investigate the anti-inflammatory effect of procyanidin B1 and the underlied mechanisms. Methods This study performed in human acute monocytic leukemia cell line (THP-1) cells, excluding lipopolysaccharide (LPS) in phosphate buffer solution (PBS) with THP-1 cells as the control group, the THP-1 cell activity in different concentration of procyanidins B1 (50, 100, 150, 200 mg / L) were detected and compared. THP-1 cells were treated with 1 mg/L LPS (LPS group), 1 mg/L LPS joint 50 mg/L procyanidin B1 (group A), 1 mg/L LPS joint 100 mg/L procyanidin B1 (group B), respectively. The levels of tumor necrosis factor-alpha (TNF-α) in the supernatants were examined by double-antibody sandwich avidin-biotin-pcroxidase complex enzyme-linked immunosorbant assay. The expression of MyD88 mRNA and myeloid differential protein 2 (MD-2) mRNA were performed by real-time PCR. Results The cell viability of procyanidins B1 in different concentration were significantly different as compared with the control group (F=31.35, P<0.001), and the cell viability of procyanidins B1 with 50, 100 mg / L showed no significant differences as compared with the control group (all P>0.05), the cell viability of procyanidins B1 with 150 and 200 mg/L were much lower than those in the control group (all P<0.05). The levels of TNF-α in group A and group B were lower than those in the LPS group, and was the lowest in the group B (all P < 0.05). The expression of MyD88 mRNA and MD-2 mRNA in the control group, LPS group and group A all showed significant difference (F=62.500, 46.700, all P<0.001), and were much higher in the LPS group than those in the control group and group A, and were higher in the group A than those in the control group (all P<0.05). Conclusion Procyanidin B1 can inhibit LPS-induced inflammatory response through negative modulation Toll-like receptor-MyD88 signaling pathway.