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
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篇名: 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
TITLE: Identification of chemical constituents in Shenfu injection and study on anti-inflammatory activities of its poly- acetylene compounds
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Westernblot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Rg(11)、人参皂苷Re(2)、人参皂苷Rb(13)、人参皂苷Rk(14)、20(R)-人参皂苷Rh(15)、20(S)-人参皂苷Rg(36)、三七皂苷R(17)、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。
ABSTRACT: OBJECTIVE To study the chemical constituents in S henfu 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 Rg3 (6),notoginsenoside R 1(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 RAW 264.7 cells;when the concentration of compound 10 ranged 12.5-50.0 μmol/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 co mpound 10 was 50.0 μmol/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.
期刊: 2022年第33卷第16期
作者: 郑琪,王晓雅,刘杰,彭成,熊亮,朱雅宁,周勤梅
AUTHORS: ZHENG Qi,WANG Xiaoya ,LIU Jie,PENG Cheng ,XIONG Liang,ZHU Yaning ,ZHOU Qinmei
关键字: 参附注射液;炔醇类;抗炎活性;人参皂苷;分离鉴定
KEYWORDS: Shenfu injection ; polyacetylenes; anti-
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