摘要
目的通过体外实验探究黑果小檗根(Berberis heteropoda Schrenk roots)不同极性部位的肠道解痉作用和抗炎活性;通过成分对比分析,初步推测潜在活性组分。方法(1)用60%乙醇和水分别提取黑果小檗根的主要成分,再对乙醇提取物依次进行系统溶剂萃取(石油醚、二氯甲烷、乙酸乙酯和饱和正丁醇),得到不同极性萃取物。(2)采用高效凝相色谱法(HPLC法)对不同极性提取物中代表性组分进行定量分析并比较其含量差异。(3)通过小鼠离体结肠平滑肌实验,研究不同提取物对正常和痉挛状态下肠道肌肉张力的影响,筛选具有抑制肠痉挛作用的活性部位。(4)采用脂多糖(LPS)诱导RAW 264.7细胞建立体外炎症细胞模型,通过不同提取物干预后测定细胞上清液中一氧化氮(NO)和白介素-6(IL-6)含量,筛选抗炎活性部位。(5)采用半制备液相色谱法对潜在活性部位进行成分分离,并用核磁共振法和质谱法进行成分定性。结果(1)HPLC成分对比分析表明,饱和正丁醇萃取物中木兰花碱、非洲防己碱、药根碱的含量高于其他萃取物,二氯甲烷萃取物中小檗胺、阿魏酸、巴马汀和小檗碱的含量高于其他萃取物。此外,发现乙酸乙酯萃取物中含有明显区别于其他萃取物的的未知峰(峰面积占比60%)。(2)小鼠结肠张力的体外实验结果表明,二氯甲烷萃取物对正常小鼠结肠张力有明显的抑制作用。此外,二氯甲烷萃取物可缓解乙酰胆碱和氯化钡引起的肠痉挛。(3)炎症细胞模型干预实验的结果表明,乙酸乙酯萃取物的抗炎作用明显优于其他萃取物。(4)从乙酸乙酯萃取物中分离得到了先前发现的未知峰化合物,结果鉴定为阿魏酸-4-O-葡萄糖苷。结论研究发现黑果小檗根二氯甲烷部位同比其他部位的肠道解痉作用最强,同时乙酸乙酯部位是发挥抗炎作用的主要活性部位,以此丰富了黑果小檗根已知的药理作用。此外,从黑果小檗根中首次分离得到了化合物:阿魏酸-4-O-葡萄糖苷,为黑果小檗根的物质基础研究提供了参考。
Objective In vitro experiments were conducted to investigate the intestinal spasmolysis and anti-inflammatory activity of different polar parts of Heiguoxiaochai(Berberis heteropoda Schrenk)roots(BHSr).The potential active components were preliminarily inferred through component comparison analysis.Method(1)The main components of BHSr were extracted with 60%ethanol and water respectively,and then the ethanol extracts were successively extracted with systematic solvent(petroleum ether,dichloromethane,ethyl acetate and saturated n-butanol).Extracts of different polarity were obtained.(2)HPLC method was used to quantitatively analyze the representative components in different polarity extracts and compare their content differences.(3)The effects of different extracts on intestinal muscle tension under normal and spasmodic conditions were studied by colon smooth muscle test in mice in vitro,and the active sites with inhibitory effects on intestinal spasmodic effects were screened.(4)Raw 264.7 cells were induced by lipopolysaccharide(LPS)to establish an inflammatory cell model in vitro.The contents of NO and IL-6 in the supernatant of cells were determined after the intervention of different extracts,and the anti-inflammatory sites were screened.(5)Semi-preparative liquid chromatography was used to separate the components of the potential active sites,and NMR and mass spectrometry were used to characterize the components.Result(1)HPLC analysis showed that the contents of magnoflorine,columbamine and jateorhizine in the saturated n-butanol extract were higher than those of other extracts,and the contents of berbamine,ferulic acid,palmatine and berberine in the dichloromethane extract were higher than those of other extracts.In addition,it was found that the ethyl acetate extract contained an unknown peak(peak area accounted for 60%)which was significantly different from other extracts.(2)The results of in vitro experiments on the colon tension of mice showed that dichloromethane extract had obvious inhibitory effect on the colon tension of normal mice.In addition,dichloromethane extracts can relieve intestinal spasms caused by acetylcholine and barium chloride.(3)The results of the inflammatory cell model intervention experiment showed that the anti-inflammatory effect of ethyl acetate extract was obviously better than that of the other extracts.(4)The unknown peak compound was isolated from the ethyl acetate extract and identified as ferulic acid 4-O-glucoside.Conclusion This study enriched the known pharmacological effects of BHSr,and provided a scientific basis for its intestinal spasmolysis and anti-inflammatory activity.In addition,the potential active components of BHSr were preliminarily explored and the compound ferulic acid-4-O-glucoside was isolated for the first time.
作者
闫琦
李莉
李松雅
冯敏
李玮
王晓梅
李飞
YAN Qi;LI Li;LI Songya;FENG Min;LI Wei;WANG Xiaomei;LI Fei(College of Pharmacy,Xinjiang Medical University,Urumqi 830054,Xinjiang,China;Shihezi University School of Medicine,Shihezi 832000,Xinjiang,China;State Key Laboratory of Bioactive Substance and Function of Natural Medicines,China Pharmaceutical University,Nanjing 210009,Jiangsu,China)
出处
《中华中医药学刊》
CAS
北大核心
2022年第3期115-120,270,共7页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(U1703108)
作者简介
闫琦(1995-),男,河南正阳人,实验师,硕士,研究方向:药物分析学;通讯作者:李莉(1963-),女,新疆乌鲁木齐人,教授,博士研究生导师,博士,研究方向:药物分析学。E-mail:llxjmu@163.com。
