目的 對RAW 264.7細胞炎癥模型進行優(yōu)化，并考察異喹啉類生物堿的抗炎活性。方法 采用內毒素脂多糖（LPS）誘導RAW264.7細胞炎癥反應，以細胞上清液中炎癥因子分泌水平為指標，優(yōu)化LPS誘導濃度、時間及陽性藥地塞米松（DEX）孵育時間，并考察異喹啉類生物堿的抗炎活性。結果 RAW 264.7細胞上清液中腫瘤壞死因子-α（TNF-α）分泌水平隨LPS誘導濃度、時間增加而升高，但當LPS質量濃度超過100 ng/mL、誘導時間超過12 h后，TNF-α分泌水平趨于平緩；LPS為20 ng/mL、誘導12 h的TNF-α分泌水平明顯升高。DEX對TNF-α分泌抑制作用隨孵育時間延長而增強，但孵育4 h后，TNF-α分泌抑制作用趨于平緩。異喹啉類生物堿呈現(xiàn)不同程度TNF-α分泌抑制作用，其抑制活性與其季胺型母核結構及R基團烷氧基取代等密切相關。結論 LPS為20 ng/mL、誘導12 h能較好誘導RAW 264.7細胞炎癥反應；異喹啉類生物堿的TNF-α分泌抑制作用與其季胺型母核結構、R基團取代等密切相關。

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.

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中國博士后科學基金項目（2019M653087）；廣東省深圳市科技計劃項目（JCYJ20170413173747440、20180306174903174、ZDSYS201707281432317、CKFW2016082916015476）；山西中醫(yī)藥大學科技創(chuàng)新能力培育計劃項目（2019PY-110、2020PY-YC-05）