CAJ | 학술논문

目的：探讨1，3－二环戊基－1，2，3，6－四氢嘧啶－4，5－二甲酸二乙酯（ZL－5015）对内毒素攻击小鼠的保护作用及机制。方法：腹腔注射脂多糖（70 mg／kg）制备小鼠内毒素中毒死亡模型和内毒素血症模型。脂多糖（10 mg／L）刺激小鼠腹腔巨噬细胞产生炎症相关细胞因子作为体外炎症模型。 ELISA法检测细胞白细胞介素1β（ IL－1β）、白细胞介素10（IL－10）和肿瘤坏死因子α（TNF－α）的含量。 Real－time PCR检测细胞因子mRNA的表达水平。结果：ZL－5015（100和200 mg／kg）预防性灌胃给药能小幅提高内毒素中毒小鼠的存活率和存活时间，在中毒早期能降低内毒素血症小鼠血清中IL－1β和TNF－α的水平，提高IL－10的水平。体外实验表明， ZL－5015（10、20和40μmol／L）能抑制内毒素刺激的腹腔巨噬细胞IL－1β和TNF－α蛋白及mRNA的表达，促进IL－10蛋白及mRNA的表达。结论：四氢嘧啶类化合物ZL－5015能提高内毒素中毒小鼠的存活率和存活时间，但作用较弱，其作用机制可能与抑制促炎细胞因子IL－1β和TNF－α、促进抑炎细胞因子IL－10的表达有关。
목적：탐토1，3－이배무기－1，2，3，6－사경밀정－4，5－이갑산이을지（ZL－5015）대내독소공격소서적보호작용급궤제。방법：복강주사지다당（70 mg／kg）제비소서내독소중독사망모형화내독소혈증모형。지다당（10 mg／L）자격소서복강거서세포산생염증상관세포인자작위체외염증모형。 ELISA법검측세포백세포개소1β（ IL－1β）、백세포개소10（IL－10）화종류배사인자α（TNF－α）적함량。 Real－time PCR검측세포인자mRNA적표체수평。결과：ZL－5015（100화200 mg／kg）예방성관위급약능소폭제고내독소중독소서적존활솔화존활시간，재중독조기능강저내독소혈증소서혈청중IL－1β화TNF－α적수평，제고IL－10적수평。체외실험표명， ZL－5015（10、20화40μmol／L）능억제내독소자격적복강거서세포IL－1β화TNF－α단백급mRNA적표체，촉진IL－10단백급mRNA적표체。결론：사경밀정류화합물ZL－5015능제고내독소중독소서적존활솔화존활시간，단작용교약，기작용궤제가능여억제촉염세포인자IL－1β화TNF－α、촉진억염세포인자IL－10적표체유관。
[ ABSTRACT] AIM:To investigate the protective effect of 1, 3-dicyclopentyl-1, 2, 3, 6-tetrahydropyrimidine-4, 5-dicarboxylic acid diethyl ester (ZL-5015) on lethal endotoxin-challenged mice and to explore the underlying mechanism. METHODS:Mouse model of lethal endotoxin challenge and endotoxemia were established by intraperitoneal administration of lipopolysaccharide (LPS) at a dose of 70 mg/kg to the C57BL/6J mice.Mouse peritoneal macrophages stimulated with LPS (10 mg/L) were used as an in vitro inflammatory model.The levels of interleukin-1β( IL-1β) , interleukin-10 ( IL-10) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).Real-time PCR was used to evaluate the mRNA expression of the cytokines.RESULTS:Prophylactic treatment of the mice with ZL-5015 (100 and 200 mg/kg, ig) slightly increased the survival rate, extended the survival time, decreased the serum levels of IL-1βand TNF-α, and increased the serum level of IL-10 in the early stage of endotoxemia as compared with model group.The results of in vitro study demonstrated that treatment of the endotoxin-stimulated mouse peritoneal macrophages with ZL-5015 (10, 20 and 40μmol/L) inhibited the expression of IL-1βand TNF-αat both mRNA and protein levels but promoted the expression of IL-10 at both mRNA and protein levels.CONCLUSION: The tetrahydropyrimidine derivative ZL-5015 shows a moderate anti-endotoxin effect by increasing the survival rate and extending the survival time of the mice challenged by endotoxin, which may result from inhibition of the expression of pro-inflammatory cytokines such as IL-1βand TNF-α, and promotion of the expression of anti-inflammatory cytokine IL-10.