参附注射液化学成分鉴定及其炔醇类化合物的抗炎活性研究  

Identification of chemical constituents in Shenfu injection and study on anti-inflammatory activities of its polyacetylene compounds

作　　者：郑琪 王晓雅 刘杰 彭成[1] 熊亮[1,2] 朱雅宁 周勤梅 ZHENG Qi;WANG Xiaoya;LIU Jie;PENG Cheng;XIONG Liang;ZHU Yaning;ZHOU Qinmei(State Key Laboratory of Southwestern Chinese Medicine Resources/School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Institute of Innovative Medicine Ingredients of Southwest Specialty Medicinal Materials,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Innovative Institute of Chinese Medicine and Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;China Resources Sanjiu(Ya’an)Pharmaceutical Co.,Ltd.,Sichuan Ya’an 625000,China)

机构地区：[1]成都中医药大学药学院/省部共建西南特色中药资源国家重点实验室,成都611137 [2]成都中医药大学西南特色药材创新药物成分研究所,成都611137 [3]成都中医药大学中医药创新研究院,成都611137 [4]华润三九(雅安)药业有限公司,四川雅安625000

出　　处：《中国药房》2022年第16期1931-1936,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81630101);四川省科技计划项目(No.2018JZ0081)。

摘　　要：目的研究参附注射液中的化学成分及其炔醇类化合物的抗炎活性。方法经大孔吸附树脂、中压液相色谱、制备薄层色谱和反相半制备高效液相色谱对参附注射液进行分离纯化,根据理化性质和波谱数据鉴定化合物结构;通过RAW264.7细胞炎症模型评价其中炔醇类化合物的抗炎活性,进一步采用Western blot法评价活性炔醇类化合物对环氧合酶-2(COX-2)蛋白表达的影响。结果从参附注射液中共分离鉴定出12个化合物,其中包括8个人参皂苷类化合物,即人参皂苷Rg1(1)、人参皂苷Re(2)、人参皂苷Rb_(1)(3)、人参皂苷Rk_(1)(4)、20(R)-人参皂苷Rh_(1)(5)、20(S)-人参皂苷Rg_(3)(6)、三七皂苷R_(1)(7)、人参三醇(8);4个炔醇类化合物,即(3R,9R,10R)-人参炔三醇(9)、人参环氧炔醇(10)、heptadeca-1,8-dien-4,6-diyne-3,10-diol(11)、人参炔醇(12)。4种炔醇类化合物中,仅化合物10有抗炎活性。化合物10对正常的RAW264.7细胞无毒;其给药浓度为12.5~50.0μmol/L时,可显著逆转脂多糖诱导的细胞上清液中NO含量的升高(P<0.05或P<0.01);在给药浓度为50.0μmol/L时,可显著逆转脂多糖诱导的细胞中COX-2蛋白表达量的升高(P<0.05)。结论化合物4、7、10~12首次在参附注射液中被鉴定并报道,其中人参环氧炔醇具有一定的抗炎作用。OBJECTIVE To study the chemical constituents in Shenfu injection and the anti-inflammatory activities of its polyacetylene compounds.METHODS Shenfu injection was separated and purified by macroporous adsorption resin,medium pressure liquid chromatography,preparative thin layer chromatography and reversed-phase semi-preparative high-performance liquid chromatography,and the compound structure was identified according to the physicochemical properties and spectral data.RAW 264.7 cell inflammation model was used to evaluate the anti-inflammatory activities of polyacetylene compounds.The effects of active polyacetylene compounds on the expressions of cyclooxygenase-2(COX-2)protein were evaluated by Western blot assay.RESULTS Twelves compounds were isolated and identified from Shenfu injection,including 8 ginsenoside compounds,i.e.ginsenoside Rg_(1)(1),ginsenoside Re(2),ginsenoside Rb_(1)(3),ginsenoside Rk_(1)(4),20(R)-ginsenoside Rh_(1)(5),20(S)-ginsenoside Rg_(3)(6),notoginsenoside R1(7),panaxatriol(8);4 polyacetylene compounds,i.e.(3R,9R,10R)-panaxytriol(9),panaxydol(10),heptadeca-1,8-dien-4,6-diyne-3,10-diol(11)and panaxynol(12).Among 4 polyacetylene compounds,only compound 10 had anti-inflammatory activity.Compound 10 was not toxic to normal RAW264.7 cells;when the concentration of compound 10 ranged 12.5-50.0μmol/L,it could significantly reverse the lipopolysaccharide-induced NO content increase in cell supernatant(P<0.05 or P<0.01);when the concentration of compound 10 was 50.0μmol/L,it could significantly reverse the lipopolysaccharide-induced protein expression increase of COX-2 in cells(P<0.05).CONCLUSIONS Compounds 4,7,10-12 are identified and reported in Shenfu injection for the first time,and panaxydol possesses a certain anti-inflammatory effect.

关 键 词：参附注射液 炔醇类 抗炎活性 人参皂苷 分离鉴定 

分 类 号：R917[医药卫生—药物分析学] R965[医药卫生—药学]