青藤碱-4-羟基-棕榈酸酯的合成及其抗炎作用

Synthesis of 4-palmitoyl-sinomenine and its anti-inflammation activity

目的合成并初步探索青藤碱衍生物青藤碱-4-羟基-棕榈酸酯的抗炎作用。方法以青藤碱(SN)为原料,化学合成青藤碱-4-羟基-棕榈酸酯。用脂多糖(LPS)诱导建立内毒素血症小鼠模型,并在诱模前于治疗组小鼠体内分别注射5 mg/kg、2.5 mg/kg剂量的青藤碱和青藤碱-4-羟基-棕榈酸酯,未治疗组和空白对照组注射生理盐水,观察记录各组小鼠的状态及生存状况。MTT法检测不同浓度青藤碱-4-羟基-棕榈酸酯体外对RAW264.7细胞增殖的影响。用终浓度为(0、1、2、5、10)μmol/L的青藤碱-4-羟基-棕榈酸酯共培养刺激RAW264.7细胞24 h,再加终浓度为1μg/kg的LPS刺激6 h后,实时定量PCR检测IL-6mRNA表达水平。结果成功制备青藤碱-4-羟基-棕榈酸酯。体内研究发现:相同浓度青藤碱-4-羟基-棕榈酸酯治疗组小鼠,生存状态较青藤碱治疗组有所改善;青藤碱-4-羟基-棕榈酸酯体外可以抑制巨噬细胞的增殖和IL-6的表达。结论青藤碱-4-羟基-棕榈酸酯具有一定的抗炎作用,可能是通过抑制巨噬细胞增殖、减少炎症介质释放而实现的。

Objective To synthesize the derivatives of sinomenine, 4-palmitoyl-sinomenine, and explore its therapeutic effect on lipopolysaccharide （ LPS）-induced endotoxemia. Methods A highly efficient synthesis of sinomenine derivatives called 4-palmitoyl-sinomenine was made with a molecule of palmitic acid substitutions at C-16 position of ring A. One hour before endotoxemia induction by i.p. injection of 10 mg/kg LPS, high-dose treatment mice （ n = 5/group） received an i. p. injection of 5 mg/kg sinomenine or 4-palmitoyl-sinomenine while the low-dose treatment mice （n=5/group） received 2.5 rng/kg sinomenine or 4-palmitoyl-sinomenine. Untreated group and normal control group received normal saline. And their survival was monitored hourly for 24 hours. Examination of cytotoxicity of 4-palmitoyl-sinomenine on RAW264. 7 cells was conducted at a concentration range of 1 to 125 pmoVL using MTT assay. RAW264. 7 cells were exposed to 4-palmitoyl-sinomenine （0, l, 2, 5, l0 μmol/L） for 24 hours, and then treated with LPS （1 μg/mL） for 6 hours. Then RAW204. ？ cells were collected and the mRNA level of IL-6 was detected by real-time quantitative PCR（qRT-PCR）. Results Sinomenine derivatives were successfully synthesized to get 4-palmitoyl-sinomenine. The survival percentage of 4-palmitoyl-sinomenine treatment groups was higher than that of sinomenine treatment groups at the same treatment concentration. The 4-palmitoyl-sinomenine inhibited RAW264. ？ cell proliferation and IL-6 gene transcription. Conclusion The 4-palmitoyl-sinomenine has an anti-inflammation probably through inhibiting the proliferation of RAW204. 7 cells and decreasing the inflammatory gene expression and inflammatory cytokine release.