目的 对RAW 264.7细胞炎症模型进行优化，并考察异喹啉类生物碱的抗炎活性。方法 采用内毒素脂多糖（LPS）诱导RAW264.7细胞炎症反应，以细胞上清液中炎症因子分泌水平为指标，优化LPS诱导浓度、时间及阳性药地塞米松（DEX）孵育时间，并考察异喹啉类生物碱的抗炎活性。结果 RAW 264.7细胞上清液中肿瘤坏死因子-α（TNF-α）分泌水平随LPS诱导浓度、时间增加而升高，但当LPS质量浓度超过100 ng/mL、诱导时间超过12 h后，TNF-α分泌水平趋于平缓；LPS为20 ng/mL、诱导12 h的TNF-α分泌水平明显升高。DEX对TNF-α分泌抑制作用随孵育时间延长而增强，但孵育4 h后，TNF-α分泌抑制作用趋于平缓。异喹啉类生物碱呈现不同程度TNF-α分泌抑制作用，其抑制活性与其季胺型母核结构及R基团烷氧基取代等密切相关。结论 LPS为20 ng/mL、诱导12 h能较好诱导RAW 264.7细胞炎症反应；异喹啉类生物碱的TNF-α分泌抑制作用与其季胺型母核结构、R基团取代等密切相关。

Objective To optimize the RAW 264.7 cell inflammation model and investigate the anti-inflammatory activity of isoquinoline alkaloids. Methods Endotoxin lipopolysaccharide (LPS) was used to induce an inflammatory response in RAW 264.7 cells, in which the induction concentration and time of LPS, and incubation time of positive drug dexamethasone (DEX) were optimized by determining the secretion level of inflammatory factors in cell supernatant. In addition, the anti-inflammatory activity of isoquinoline alkaloids were investigated. Results The secretion level of TNF-αin supernatant of RAW 264.7 cells increased with the inductive concentration and time of LPS induction, while the level of TNF-α tended to be flat when the induction concentration of LPS exceeded 100 ng/mL, or the induction time exceeded 12 h. In addition, the secretion level of TNF-α was higher when the induction concentration and time of LPS were 20 ng/mL and 12 h, respectively. Besides, the inhibitory effect of DEX on TNF-α increased with its incubation time, while the inhibitory effect of DEX on TNF-α secretion tended to be flat after incubation for 4 h. The anti-inflammatory results showed that the isoquinoline alkaloids exhibited different degrees of inhibition on TNF-α secretion, and the anti-inflammatory activity were closely correlated with their quaternary amine parent nucleus and alkoxy substitution of R group. Conclusion LPS at 20 ng/mL for 12 h incubation could better induce the inflammatory response in RAW 264.7 cells, and the inhibitory effect of isoquinoline alkaloids on the secretion of TNF-α are closely related to their quaternary amine parent nucleus structure and R group substitution.

R285.5

中国博士后科学基金项目（2019M653087）；广东省深圳市科技计划项目（JCYJ20170413173747440、20180306174903174、ZDSYS201707281432317、CKFW2016082916015476）；山西中医药大学科技创新能力培育计划项目（2019PY-110、2020PY-YC-05）