趙鵬安,力瓦衣丁·買合蘇提,馬國需, ,孫照翠,許旭東,石磊嶺*,高興旺
大苞荊芥化學(xué)成分及其抗炎活性研究
趙鵬安1, 2,力瓦衣丁·買合蘇提2,馬國需2, 3,孫照翠3,許旭東3,石磊嶺2*,高興旺1*
1. 新疆大學(xué),新疆 烏魯木齊 830046 2. 新疆維吾爾自治區(qū)中藥民族藥研究所,新疆 烏魯木齊 830002 3. 中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)院藥用植物研究所,北京 100193
闡明維藥大苞荊芥中的活性物質(zhì)并研究其抗炎活性。采用多種色譜方法對大苞荊芥化學(xué)成分進行系統(tǒng)的分離純化,并利用波譜解析和理化性質(zhì)對化合物的結(jié)構(gòu)進行鑒定。同時采用MTT比色法,以氫化可的松作為陽性對照,對化合物進行體外抗炎活性實驗。從大苞荊芥中分離鑒定了20個化合物(包括2個酚酸類化合物和18個二萜類化合物),分別為7-氧代去氫松香酸(1)、脫氫松香醇(2)、朗伯酸(3)、3β,7α-二羥基-松香-8,11,13-三烯(4)、雷酚萜L(5)、13-hydroxy-9-oxo-9,10-seco-abiet-8(14)-en-18,10α-olide(6)、8,11,13-阿松香三烯-7a,18-二醇(7)、3β,19-二羥基-松香-8,11,13-三烯(8)、7α,15,18-三羥基-松香-8,11,13-三烯(9)、松香烷-8,11,13-三烯-7β-醇(10)、6α-羥基脫氫松香酸(11)、15-羥基-松香-8,11,13-三烯-19-乙酸酯(12)、7a-羥基-松香-8,11,13-三烯-19-乙酸酯(13)、水楊酸(14)、反式-3-甲氧基-4-羥基肉桂酸(15)、15-羥基松香酸(16)、7β-羥基脫氫松香酸(17)、6-羥基脫氫松香酸(18)、角果酸G(19)、家地非諾酸K(20)??寡谆钚詫嶒灲Y(jié)果表明,化合物11對脂多糖(lipopolysaccharide,LPS)誘導(dǎo)的小鼠巨噬細胞系RAW264.7中NO釋放有抑制作用,半數(shù)抑制濃度(median inhibition concentration,IC50)值為0.256 mg/mL。化合物5、6、11、13為首次從唇形科植物中分離得到;化合物1、3、4、7、8、10、12為首次從荊芥屬植物中分離得到;化合物2、9、14、15為首次從大苞荊芥中分離得到;化合物11具有一定的抗炎活性,具有進一步深入研究的價值。
大苞荊芥;二萜類化合物;抗炎活性;雷酚萜L;6α-羥基脫氫松香酸;7a-羥基-松香-8,11,13-三烯-19-乙酸酯
炎癥是具有血管系統(tǒng)的活體組織面對損傷性刺激,如感染、組織損傷等,所發(fā)生的復(fù)雜防御反應(yīng)[1]。病毒和細菌是誘導(dǎo)炎癥反應(yīng)的主要介質(zhì),物理因素、化學(xué)因素以及創(chuàng)傷都會對這一反應(yīng)產(chǎn)生影響[2-3]。發(fā)炎是身體對抗細菌和病毒所致感染,長時間或過度炎癥在體內(nèi)引發(fā)的一系列異常反應(yīng),引起各種疾?。ㄈ缣悄虿?、哮喘和瘧疾、類風(fēng)濕性關(guān)節(jié)炎),在嚴(yán)重的情況下甚至出現(xiàn)多器官功能障礙綜合征,威脅人類健康[4]。因此,不斷地研究安全有效的抗炎藥是一項長期任務(wù)。Byun等[5]研究發(fā)現(xiàn),荊芥的乙醇提取物能抑制脂多糖(lipopolysaccharide,LPS)誘導(dǎo)的細胞表面分子(CD80和CD86)的表達和促炎細胞因子,如腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)、白細胞介素-1β (Interleukin-1β,IL-1β)、IL-6的產(chǎn)生。Choi等[6]研究發(fā)現(xiàn),荊芥可降低特異性皮炎模型小鼠血清中的免疫球蛋白E(immunoglobulin E,IgE)、TNF-α、IL-6的含量。Zhang等[7]研究發(fā)現(xiàn)二萜類是荊芥屬植物發(fā)揮臨床抗炎作用的物質(zhì)基礎(chǔ)。Yang等[8]通過研究發(fā)現(xiàn)松香烷型二萜類化合物的是大苞荊芥在臨床上發(fā)揮抗炎作用的物質(zhì)基礎(chǔ),值得進一步研究。
大苞荊芥Benth.為唇形科(Labiatae)荊芥屬Linn.植物,主要分布于巴基斯坦、尼泊爾、伊朗等國家[9-10],是我國一種主要依靠進口的民間習(xí)用植物藥材,維吾爾名為“祖發(fā)”。據(jù)《維吾爾藥志》記載,大苞荊芥全草含揮發(fā)油、黃酮類、多糖類、萜類、有機酸類等化學(xué)成分,具有鎮(zhèn)咳平喘、清熱利濕之功效,用于氣管炎、咳嗽氣喘、感冒發(fā)燒等癥[11]。大苞荊芥常被作為神香草的替代品在民醫(yī)中廣泛用使用,因而對神香草的了解和研究較多,對大苞荊芥的研究較少,缺乏對大苞荊芥化學(xué)成分及其藥用活性的系統(tǒng)研究。因此本研究選擇大苞荊芥作為研究對象,應(yīng)用薄層色譜、硅膠柱色譜、反相C18柱色譜以及半制備型HPLC分別對大苞荊芥95%乙醇提取液的二氯甲烷和石油醚萃取部位進行了分離純化,針對大苞荊芥活性部位中的二萜類活性成分進行系統(tǒng)研究,通過核磁共振手段結(jié)合性狀特征鑒定化合物結(jié)構(gòu),共鑒定20個化合物,包括18個二萜類化合物,分別為7-氧代去氫松香酸(7-oxodehydroabietic acid,1)、脫氫樅香醇(dehydroabietol,2)、朗伯酸(lambertic acid,3)、3β,7α-二羥基-松香-8,11,13-三烯(3β,7α-dihydroxy- abieta-8,11,13- triene,4)、雷酚萜L(triptobenzene L,5)、13-hydroxy-9-oxo-9,10--abiet-8(14)-en-18,10a- olide(6)、8,11,13-阿松香三烯-7a,18-二醇(8,11,13- abietatriene-7a,18-diol,7)、3β,19-二羥基-松香- 8,11,13-三烯(3β,19-dihydroxy-abieta-8,11,13-triene,8)、7α,15,18-三羥基-松香-8,11,13-三烯(7α,15, 18-trihydroxyabieta-8,11,13-triene,9)、松香烷- 8,11,13-三烯-7β-醇(abieta-8,11,13-trien-7β-ol,10)、6α-羥基脫氫松香酸(6α-hydroxycallitrisic acid,11)、15-羥基-松香-8,11,13-三烯-19-乙酸酯(19-acetoxy- 15-hydroxyabieta-8,11,13-triene,12)、7a-羥基-松香- 8,11,13-三烯-19-乙酸酯(7a-hydroxyabieta-8,11,13- trien-19-ylacetate,13)、水楊酸(salicylis acid,14)、反式-3-甲氧基-4-羥基肉桂酸(-3-hydroxy-4- methoxy-cinnamic acid,15)、15-羥基松香酸(15-hydroxyabieta-8,11,13-trien-18-oic acid,16)、7β-羥基脫氫松香酸(7β-hydroxycallitrisic acid,17)、6-羥基脫氫松香酸(6-hydroxycallitrisic acid,18)、角果酸G(angustanoic acid G,19)、家地非諾酸K(jiadifenoic acid K,20)?;衔?、6、11、13為首次從唇形科植物中分離得到;化合物1、3、4、7、8、10、12為首次從荊芥屬植物中分離得到;化合物2、9、14、15為首次從大苞荊芥中分離得到。對分離得到的二萜類化合物進行了體外MTT法抗炎活性測試,其中化合物1、2、11、18對LPS誘導(dǎo)的RAW264.7巨噬細胞NO的釋放均具有不同程度的抑制作用,其中化合物11的抑制作用相對較好,半數(shù)抑制濃度(median inhibition concentration,IC50)值為0.256 mg/mL。本研究為民間藥大苞荊芥的臨床應(yīng)用提供質(zhì)量保障和參考依據(jù),也為大苞荊芥的二次開發(fā)和推廣應(yīng)用奠定科學(xué)基礎(chǔ)。
R-1005雙回流旋轉(zhuǎn)蒸發(fā)儀(方圓儀器科技有限公司);BrukerAvanceIII600型NMR(德國布魯克公司);KH-700DB臺式數(shù)控超聲波清洗器(蘇州安源儀器有限公司);BYLABUVIII紫外燈(諾雷信達科技有限公司);半制備型液相色譜儀(硅儀生化科技有限公司);乙醇、甲醇、石油醚、二氯甲烷(琳盛化工有限公司);柱色譜硅膠(創(chuàng)益化工有限公司);薄層色譜硅膠HF254(永德茂科技有限公司);Rp-C18(北京譜朋科技有限公司);純凈水(福萬家純凈水有限公司);氫化可的松(批號B21001,上海源葉生物有限公司)。
藥材于2020年購自于新疆維吾爾自治區(qū)麥迪森藥業(yè)有限公司,經(jīng)新疆維吾爾自治區(qū)藥品檢驗研究院蘇萊曼·哈力克主任藥師鑒定為大苞荊芥Benth.的干燥全草,憑證藥材標(biāo)本(M20191025)保存于新疆中藥民族藥研究所藥材室。
實驗所用小鼠單核巨噬細胞RAW264.7購自中國醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所基礎(chǔ)醫(yī)學(xué)中心。
取6 kg大苞荊芥全草打粉,用95%乙醇加熱回流提取3次,每次2 h。合并提取液,濃縮后得浸膏437 g。將浸膏用水分散后依次用石油醚和二氯甲烷萃取,萃取液濃縮后得石油醚部位134.8 g、二氯甲烷部位67.4 g。
石油醚部位經(jīng)100~200目硅膠色譜分離,石油醚-醋酸乙酯(90∶1→10∶1)洗脫后得7個流分Fr. B1~B7。Fr. B6(12.0 g)依次用硅膠2次分離得4個流分Fr. B6-3a~B6-3d。Fr. B6-3a(3.0 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,90%甲醇,2.0 mL/min)分離得化合物1(3.6 mg)。Fr. B6-3b(2.1 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,85%甲醇,2.0 mL/min)分離得化合物2(3.6 mg)。Fr. B6-3d(1.2 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,85%甲醇,2.0 mL/min)分離得化合物3(2.5 mg)。Fr. B7(23.0 g)用硅膠柱得Fr. B7-1~B7-5共5個部位。Fr. B7-3(10.0 g)用硅膠柱洗脫,得4個流分Fr. B7-3a~B7-3d。Fr. B7-3a(2.8 g)用制備液相(HPLC-5C18E,250 mm×10 mm,3 μm,85%甲醇,2.0 mL/min)得化合物4(2.0 mg)、5(4.2 mg)。Fr. B7-3b(1.5 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,75%甲醇,2.0 mL/min)得到化合物6(2.0 mg)、7(2.8 mg)。Fr. B7-3c(1.6 g)用制備液相色譜分離(HPLC-5C18E,250 mm×10 mm,3 μm,80%甲醇,2.0 mL/min)得化合物8(2.5 mg)、9(3.3 mg)、10(3.7 mg)。Fr. B7-2(14.3 g)用硅膠柱得4個部位Fr. B7-2a~B7-2d。Fr. B7-2a(4.0 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,90%甲醇,2.0 mL/min)分離得化合物11(4.1 mg)、12(2.9 mg)、13(3.1 mg)。
二氯甲烷部位經(jīng)100~200目硅膠柱色譜分離,二氯甲烷-甲醇(100∶1→2∶1)洗脫后得7個流分Fr. A1~A7,F(xiàn)r. A1(15.0 g)通過硅膠柱再次分離得7個部位Fr. A1-1~A1-7。Fr. A1-2(2.1 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,60%甲醇,2.0 mL/min)分離,得化合物14(3.6 mg)。Fr. A1-4(3.6 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,40%甲醇,2.0 mL/min)分離得化合物15(3.0 mg)。Fr. A1~A3(12.0 g)用C18柱分離得4個流分(Fr. A1-3a~A1-3d)。Fr. A1-3b(1.4 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,75%甲醇,2.0 mL/min)分離,得化合物16(2.0 mg)、17(2.7 mg)。Fr. A1-3c(1.1 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,38%甲醇,2.0 mL/min)分離,得化合物18(2.6 mg)。Fr. A2(10.0 g)用硅膠柱分離得6個部分 Fr. A2-1~A2-6。Fr. A2-2(2.3 g)用制備液相色譜(HPLC-5C18E,250 mm×10 mm,3 μm,85%甲醇,2.0 mL/min)分離,得化合物19(2.3 mg)、20(3.6 mg)。
首先對巨噬細胞RAW264.7進行培養(yǎng)并制成單細胞懸液,將制好的單細胞懸液,以200 μL/孔接種于96孔板內(nèi),37 ℃培養(yǎng),隨后放入5% CO2培養(yǎng)箱貼壁24 h,各組加入磷酸緩沖液,使質(zhì)量濃度為1 μg/mL,繼續(xù)培養(yǎng)24 h后離心,取上清液放于96孔板中備用;氫化可的松用含10% FBS的DMEM培養(yǎng)液稀釋,樣品在96孔板中以50 μL/孔加入。之后室溫下在每孔加50 μL的Griess Reagent I和II,混勻,室溫避光放置10 min,用酶標(biāo)儀540 nm測定吸光度()值,計算NO含量。
NO含量=1-(給藥-對照)/對照
化合物1:白色無定形粉末。ESI-MS/: 337 [M+Na]+,13C-NMR、APT譜確定分子式C20H26O3。1H-NMR (600 MHz, CDCl3): 7.90 (1H, d,= 3.6 Hz, H-14), 7.43 (1H, dd,= 1.8, 2.4 Hz, H-12), 7.34 (1H, d,= 8.4 Hz, H-11), 2.30 (1H, d,= 13.2 Hz, H-5), 2.19 (1H, m, H-2b), 2.11 (1H, m, H-3b), 2.10 (1H, m, H-1b), 1.73 (1H, m, H-6b), 1.66 (1H, m, H-3a), 1.55 (1H, m, H-6a), 1.46 (1H, m, H-2a), 1.43 (1H, m, H-1a), 1.32 (3H, s, H-16), 1.32 (3H, s, H-17), 1.21 (3H, s, H-19), 1.17 (1H, d,= 7.0 Hz, H-15), 0.96 (3H, d,= 7.8 Hz, H-20);13C-NMR (150 MHz, CDCl3): 199.3 (C-7), 180.6 (C-18), 152.1 (C-9), 147.1 (C-13), 132.8 (C-14), 130.6 (C-8), 129.0 (C-12), 125.0 (C-11), 50.3 (C-5), 43.8 (C-4), 38.6 (C-1), 37.7 (C-10), 37.5 (C-3), 33.8 (C-15), 30.7 (C-6), 28.3 (C-20), 24.0 (C-17), 19.8 (C-2), 13.9 (C-19)。經(jīng)數(shù)據(jù)比對[12],確定化合物1為7-氧代去脫氫松香酸。
化合物2:白色無定形粉末。ESI-MS/: 309 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O。1H-NMR (600 MHz, CDCl3): 7.18 (1H, dd,= 7.8, 4.2 Hz, H-11), 7.05 (1H, d,= 7.2 Hz, H-12), 6.93 (1H, s, H-14), 3.72 (1H, d,= 11.4 Hz, H-18b), 3.49 (1H, d,= 11.4 Hz, H-18a), 2.31 (1H, m, H-1b), 2.44 (1H, m, H-5), 2.22 (1H, m, H-3b), 2.01 (1H, m, H-6b), 1.93 (1H, m, H-7b), 1.87 (1H, m, H-2b), 1.60 (1H, m, H-3a), 1.54 (1H, m, H-2a), 1.51 (1H, m, H-6a), 1.40 (1H, m, H-1a), 1.39 (6H, s, H-16, 17), 1.21 (1H, d,= 7.2 Hz, 1.19 (1H, m, H-7a), 1.19 (3H, d,= 6.8 Hz, H-20), 1.12 (3H, s, H-19);13C-NMR (150 MHz, CDCl3): 146.5 (C-9), 144.7 (C-13), 134.9 (C-8), 129.0 (C-14), 127.2 (C-12), 124.8 (C-11), 67.2 (C-18), 51.9 (C-4), 43.8 (C-5), 37.3 (C-1), 37.3 (C-10), 36.0 (C-3), 33.7 (C-15), 33.6 (C-20), 30.1 (C-7), 30.0 (C-6), 24.8 (C-17), 24.1 (C-16), 20.0 (C-2), 16.8 (C-19)。經(jīng)數(shù)據(jù)比對[13],確定化合物2為脫氫松香醇。
化合物3:白色無定形粉末。ESI-MS/: 349 [M+Na]+,13C-NMR、APT譜確定分子式C20H28O3。1H-NMR (600 MHz, CDCl3): 6.83 (1H, s, H-14), 6.63 (1H, s, H-11), 3.21 (1H, m, H-15), 2.81 (1H, m, H-7b), 2.67 (1H, m, H-7a), 2.43 (1H, m, H-2b), 2.13 (1H, d,= 12.0 Hz, H-5), 2.19 (1H, m, H-1b), 2.11 (1H, m, H-3b), 1.68 (1H, m, H-6b), 1.65 (1H, m, H-2a), 1.59 (1H, m, H-3a), 1.43 (1H, m, H-6a), 1.35 (1H, m, H-1a), 1.32 (6H, s, H-16, 17), 1.19 (3H, d,= 7.2 Hz, H-18), 1.10 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 183.5 (C-19), 151.0 (C-12), 132.0 (C-13), 127.6 (C-8), 126.8 (C-14), 126.8 (C-9), 112.1 (C-11), 52.9 (C-5), 44.0 (C-4), 39.5 (C-10), 38.4 (C-1), 37.5 (C-3), 31.4 (C-7), 29.8 (C-6), 28.9 (C-15), 26.9 (C-18), 23.3 (C-20), 22.9 (C-17), 22.7 (C-16), 19.6 (C-2)。經(jīng)數(shù)據(jù)比對[14],確定化合物3為朗伯酸。
化合物4:白色無定形粉末。ESI-MS/: 323 [M+Na]+,13C-NMR、APT譜確定分子C20H30O2。1H-NMR (600 MHz, CDCl3): 7.13 (1H, s, H-11), 7.12 (1H, s, H-14), 7.08 (1H, s, H-12), 4.31 (1H, m, H-7), 3.37 (1H, m, H-3), 2.89 (1H, m, H-15), 1.82 (1H, m, H-1b), 1.79 (1H, m, H-6b), 1.72 (1H, m, H-2b), 1.61 (1H, m, H-5), 1.57 (1H, m, H-1a), 1.48 (1H, m, H-6a), 1.47 (1H, m, H-2a), 1.23 (3H, d,= 7.2 Hz, H-20), 1.13 (3H, s, H-16), 1.10 (3H, s, H-17), 0.94 (6H, s, H-18, 19);13C-NMR (150 MHz, CDCl3): 150.0 (C-13), 146.9 (C-9), 136.1 (C-8), 126.9 (C-14), 126.0 (C-11), 124.7 (C-12), 78.9 (C-3), 69.7 (C-7), 44.3 (C-5), 38.6 (C-10), 37.9 (C-4), 36.8 (C-1), 33.7 (C-15), 30.9 (C-6), 28.5 (C-20), 28.2 (C-2), 24.2 (C-16), 24.0 (C-17), 15.7 (C-18), 15.7 (C-19)。經(jīng)數(shù)據(jù)比對[15],確定化合4為3β,7α-二羥基-松香-8,11,13-三烯。
化合物5:白色無定形粉末。ESI-MS/: 325 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O2。1H-NMR (600 MHz, CDCl3): 7.13 (1H, d,= 7.2 Hz, H-11), 7.00 (1H, d,= 7.2 Hz, H-12), 6.89 (1H, s, H-14), 4.33 (1H, d,= 11.2 Hz, H-19b), 3.53 (1H, dd,= 11.7, 4.4 Hz, H-3), 3.43 (1H, d,= 11.2 Hz, H-19a), 2.92 (1H, m, H-7b), 2.83 (1H, m, H-7a), 2.79 (1H, d,= 8.2 Hz, H-15), 2.34 (1H, ddd,= 13.3, 3.4, 3.4 Hz, H-1b), 2.01 (1H, m, H-2b), 1.97 (1H, m, H-6b), 1.91 (1H, m, H-2a), 1.66 (1H, m, H-6a), 1.54 (1H, ddd,= 13.3, 13.3, 3.5 Hz, H-1a), 1.46 (1H, dd,= 12.3, 1.2 Hz, H-5), 1.32 (3H, s, H-18), 1.22 (6H, d,= 6.9 Hz, H-16, 17), 1.15 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 146.3 (C-9), 146.0 (C-13), 134.4 (C-8), 126.8 (C-14), 124.5 (C-11), 124.1 (C-12), 80.7 (C-3), 64.2 (C-19), 50.7 (C-5), 43.0 (C-4), 37.2 (C-10), 36.8 (C-1), 33.5 (C-15), 31.1 (C-7), 28.6 (C-2), 26.0 (C-20), 24.0 (C-16), 24.0 (C-17), 22.5 (C-18), 19.0 (C-6)。經(jīng)數(shù)據(jù)比對[16],確定化合物5為雷酚萜L。
化合物6:白色無定形粉末。ESI-MS/: 357 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O4。1H-NMR (600 MHz, CDCl3): 6.50 (1H, d,= 0.9 Hz, H-14), 2.69 (1H, ddd,= 17.2, 10.3, 5.3 Hz, H-11b), 2.44 (1H, ddd,= 17.2, 6.4, 4.8 Hz, H-11a), 2.28 (1H, dddd,= 13.5, 10.8, 5.9, 0.9 Hz, H-7b), 2.20 (1H, dddd,= 13.5, 10.8, 5.0, 0.9 Hz, H-7a), 2.14 (1H, ddd,= 13.5, 6.4, 5.3 Hz, H-12b), 1.96 (1H, ddd,= 13.5, 10.3, 4.8 Hz, H-12a), 1.92 (1H, d,= 6.9 Hz, H-15), 1.80 (1H, t,= 6.1 Hz, H -5), 1.70 (1H, m, H-1b), 1.68 (1H, m, H-2b), 1.53 (1H, m, H-2a), 1.52 (1H, m, H-3b), 1.51 (1H, m, H-6b), 1.46 (1H, m, H-6a), 1.45 (3H, s, H-20) 1.39 (1H, m, H-3a), 1.29 (1H, m, H-1a), 1.26 (3H, s, H-19), 1.01 (3H, d,= 7.2 Hz, H-16), 0.96 (3H, d,= 7.4 Hz, H-17);13C-NMR (150 MHz, CDCl3): 199.1 (C-9), 180.4 (C-18), 28.4 (C-1), 148.1 (C-14), 139.4 (C-8), 85.0 (C-10), 72.3 (C-13), 55.2 (C-5), 48.5 (C-4), 37.0 (C-15), 34.1 (C-11), 30.8 (C-12), 29.7 (C-7), 25.4 (C-3), 23.7 (C-6), 22.2 (C-20), 19.7 (C-2), 19.7 (C-19), 17.5 (C-17), 16.4 (C-16)。經(jīng)數(shù)據(jù)比對[17],確定化合物6為13-hydroxy-9-oxo-9,10--abiet-8 (14)-en-18,10a-olide。
化合物7:白色無定形粉末。ESI-MS/: 325 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O2。1H-NMR (600 MHz, CDCl3): 7.22 (1H, d,= 7.2 Hz, H-11), 7.16 (1H, s, H-12), 7.14 (1H, d,= 8.2 Hz, H-14), 3.87 (1H, d,= 11.4 Hz, H-19b), 3.55 (1H, d,= 10.8 Hz, H-19a), 3.21 (1H, m, H-7), 2.88 (1H, m, H-15), 2.61 (1H, m, H-3b), 2.35 (1H, m, H-3a), 2.22 (1H, m, H-1b), 2.07 (1H, m, H-2b), 1.99 (1H, m, H-6b), 1.92 (1H, m, H-2a), 1.71 (1H, m, H-1a), 1.61 (1H, m, H-6a), 1.30 (1H, m, H-5), 1.24 (3H, s, H-18), 1.23 (3H, s, H-16), 1.13 (3H, s, H-17), 1.08 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 147.1 (C-13), 146.5 (C-9), 135.7 (C-8), 127.7 (C-14), 126.7 (C-11), 124.6 (C-12), 68.3 (C-7), 65.5 (C-19), 45.1 (C-5), 38.5 (C-4), 37.8 (C-10), 35.2 (C-1), 33.5 (C-15), 29.7 (C-3), 28.6 (C-2), 26.7 (C-20), 24.7 (C-18), 24.1 (C-16), 23.8 (C-17), 19.0 (C-6)。經(jīng)數(shù)據(jù)比對[18],確定化合物7為8,11,13-阿松香三烯-7α,18-二醇。
化合物8:白色無定形粉末。ESI-MS/: 325 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O2。1H-NMR (600 MHz, CDCl3): 7.19 (1H, d,= 6.2 Hz, H-11), 7.17 (1H, s, H-12), 7.09 (1H, s, H-14), 4.44 (1H, d,= 11.4 Hz, H-19b), 4.22 (1H, d,= 11.8 Hz, H-19a), 3.36 (1H, dd,= 4.2, 4.8 Hz, H-3), 2.95 (1H, m, H-7b), 2.87 (1H, m, H-15), 2.37 (1H, m, H-1b), 2.28 (1H, m, H-7a), 2.23 (1H, m, H-2b), 2.09 (1H, m, H-6b), 1.76 (1H, m, H-2a), 1.63 (1H, m, H-6a), 1.62 (1H, m, H-1a), 1.56 (3H, s, H-18), 1.49 (1H, m, H-5), 1.25 (3H, s, H-16), 1.23 (3H, s, H-17), 1.20 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 147.0 (C-9), 146.3 (C-13), 134.5 (C-14), 134.4 (C-8), 124.8 (C-11), 122.3 (C-12), 78.9 (C-3), 65.3 (C-19), 51.1 (C-5), 42.3 (C-4), 37.2 (C-10), 35.4 (C-1), 34.5 (C-15), 31.6 (C-7), 28.0 (C-2), 25.4 (C-20), 22.5 (C-16), 22.5 (C-17), 19.6 (C-6), 19.5 (C-18)。經(jīng)數(shù)據(jù)比對[16],確定化合物8為3β,19-二羥基-松香-8,11,13-三烯。
化合物9:白色無定形粉末。ESI-MS/: 341 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O3。1H-NMR (600 MHz, CDCl3): 7.32 (1H, s, H-11), 6.89 (1H, s, H-12), 6.78 (1H, s, H-14), 4.43 (1H, m, H-19b), 4.18 (1H, m, H-19a), 3.23 (1H, m, H-7), 2.32 (1H, m, H-1b), 2.28 (1H, m, H-3b), 2.17 (1H, m, H-6b), 1.99 (1H, m, H-2b), 1.92 (1H, m, H-3a), 1.82 (1H, m, H-2a), 1.81 (1H, m, H-6a), 1.70 (1H, m, H-5),1.59 (1H, m, H-1a), 1.53 (3H, s, H-18), 1.36 (3H, s, H-16), 1.28 (3H, s, H-17), 1.06 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 146.8 (C-9), 144.8 (C-13), 135.7 (C-8), 127.3 (C-14), 126.1 (C-11), 125.6 (C-12), 68.7 (C-7), 66.3 (C-19), 58.3 (C-15), 45.7 (C-5), 43.6 (C-4), 38.9 (C-1), 37.5 (C-3), 37.5 (C-10), 28.6 (C-2), 28.6 (C-20), 24.2 (C-16), 24.0 (C-17), 20.0 (C-6), 16.1 (C-18)。經(jīng)數(shù)據(jù)比對[19],確定化合物9為7α,15,18-三羥基-松香-8,11,13-三烯。
化合物10:白色無定形粉末。ESI-MS/: 309 [M+Na]+,13C-NMR、APT譜確定分子式C20H30O。1H-NMR (600 MHz, CDCl3): 7.44 (1H, s, H-11), 7.14 (1H, d,= 7.2 Hz, H-12), 7.08 (1H, d,= 6.8 Hz, H-14), 2.87 (1H, s, H-15), 1.61 (1H, m, H-1a), 2.62 (1H, m, H-3b), 2.57 (1H, m, H-7), 2.56 (1H, m, H-1b), 2.36 (1H, m, H-6b), 2.33 (1H, m, H-3a), 2.03 (1H, m, H-2b), 1.99 (1H, m, H-5), 1.97 (1H, m, H-2a), 1.48 (1H, m, H-6a), 1.32 (3H, s, H-18), 1.24 (3H, s, H-16), 1.23 (3H, s, H-17), 1.21 (3H, s, H-20), 1.16 (3H, s, H-19);13C-NMR (150 MHz, CDCl3): 146.7 (C-9), 145.3 (C-13), 138.3 (C-8), 126.0 (C-14), 125.4 (C-11), 124.6 (C-12), 71.7 (C-7), 50.2 (C-5), 43.6 (C-4), 39.1 (C-10), 37.4 (C-1), 33.9 (C-15), 32.1 (C-3), 29.9 (C-2), 28.8 (C-20), 24.2 (C-16), 24.0 (C-17), 23.3 (C-18), 19.9 (C-6), 14.3 (C-19)。經(jīng)數(shù)據(jù)比對[20],確定化合物10為松香烷-8,11,13-三烯-7β-醇。
化合物11:白色無定形粉末。ESI-MS/: 339 [M+Na]+,13C-NMR、APT譜確定分子式C20H28O3。1H-NMR (600 MHz, CDCl3): 7.18 (1H, d,= 7.4 Hz, H-11), 7.00 (1H, d,= 7.8 Hz, H-12), 6.91 (1H, s, H-14), 1.54 (1H, m, H-1a), 2.81 (1H, m, H-15), 2.76 (1H, m, H-3b), 2.61 (1H, dd,= 9.0, 10.2 Hz, H-6), 2.36 (1H, m, H-7b), 2.33 (1H, m, H-3a), 2.19 (1H, m, H-5), 2.19 (1H, m, H-1b), 2.03 (1H, m, H-2b), 1.99 (1H, m, H-2a), 1.67 (1H, m, H-7a), 1.37 (3H, s, H-18), 1.23 (3H, s, H-17), 1.22 (3H, s, H-16), 1.10 (3H, s, H-19);13C-NMR (150 MHz, CDCl3): 182.6 (C-20), 146.2 (C-9), 144.6 (C-13), 134.8 (C-8), 127.1 (C-14), 125.3 (C-11), 124.4 (C-12), 65.0 (C-6), 52.2 (C-5), 44.9 (C-4), 39.6 (C-10), 33.6 (C-15), 32.0 (C-3), 31.9 (C-1), 29.7 (C-2), 24.3 (C-16), 24.1 (C-17), 24.1 (C-18), 20.7 (C-7), 14.3 (C-19)。經(jīng)數(shù)據(jù)比對[19],確定化合物11為6α-羥基脫氫松香酸。
化合物12:白色無定形粉末。ESI-MS/: 367 [M+Na]+,13C-NMR、APT譜確定分子式C22H32O3。1H-NMR (600 MHz, CDCl3): 7.39 (1H, s, H-12), 7.19 (1H, s, H-11), 7.16 (1H, s, H-14), 4.33 (1H, d,= 11.2 Hz, H-20b), 4.01 (1H, d,= 11.4 Hz, H-20a), 2.90 (1H, m, H-7b), 2.34 (1H, m, H-7a), 2.07 (3H, s, H-22), 1.82 (1H, m, H-1b), 1.72 (1H, m, H-6b), 1.57 (1H, m, H-1a), 1.56 (1H, m, H-3b), 1.55 (1H, m, H-5), 1.52 (6H, s, H-16/17), 1.43 (1H, m, H-2b), 1.43 (1H, m, H-6a), 1.21 (1H, m, H-3a), 1.21 (3H, s, H-18), 1.19 (1H, m, H-2a), 1.04 (3H, s, H-19);13C-NMR (150 MHz, CDCl3): 171.5 (C-21), 148.0 (C-9), 146.0 (C-13), 134.7 (C-8), 124.9 (C-14), 124.6 (C-11), 122.1 (C-12), 73.5 (C-20), 67.0 (C-15), 51.2 (C-5), 38.7 (C-4), 37.2 (C-10), 36.0 (C-1), 31.6 (C-16), 31.6 (C-17), 31.2 (C-3), 30.4 (C-7), 29.4 (C-2), 27.4 (C-18), 25.7 (C-19), 21.1 (C-22), 19.3 (C-6)。經(jīng)數(shù)據(jù)比對[21],確定化合物12為15-羥基-松香-8,11,13-三烯-19-乙酸酯。
化合物13:白色無定形粉末。ESI-MS/: 367 [M+Na]+,13C-NMR、APT譜確定分子式C22H32O3。1H-NMR (600 MHz, CDCl3): 7.26 (1H, s, H-12), 7.20 (1H, d,= 7.4 Hz, H-11), 7.13 (1H, d,= 2.4 Hz, H-14), 4.28 (1H, d,= 8.6 Hz, H-20b), 3.98 (1H, d,= 12.2 Hz, H-20a), 2.93 (1H, m, H-7), 2.86 (1H, m, H-15), 1.99 (3H, s, H-22), 1.82 (1H, m, H-1b), 1.72 (1H, m, H-6b), 1.67 (1H, m, H-2b), 1.60 (1H, m, H-3b), 1.54 (1H, m, H-1a), 1.54 (1H, m, H-5), 1.41 (1H, m, H-6a), 1.29 (1H, m, H-2a), 1.25 (3H, s, H-16), 1.24 (3H, s, H-17), 1.15 (1H, m, H-3a), 1.15 (3H, s, H-18), 1.07 (3H, s, H-19);13C-NMR (150 MHz, CDCl3): 171.6 (C-21), 147.1 (C-9), 146.89 (C-13), 135.9 (C-8), 36.2 (C-1), 128.0 (C-14), 126.9 (C-11), 124.9 (C-12), 79.2 (C-7), 69.9 (C-20), 45.3 (C-5), 38.7 (C-10), 38.7 (C-4), 33.8 (C-15), 32.7 (C-3), 28.7 (C-2), 27.3 (C-16), 24.9 (C-17), 24.2 (C-18), 24.1 (C-19), 21.2 (C-22), 19.1 (C-6)。經(jīng)數(shù)據(jù)比對[22],確定化合物13為7a-羥基-松香-8,11,13-三烯-19-乙酸酯。
化合物14:淡黃色油狀物。ESI-MS/: 161 [M+Na]+,13C-NMR、APT譜確定分子式C7H6O3。1H-NMR (600 MHz, CDCl3): 7.93 (1H, dd,= 8.0, 1.4 Hz, H-6), 7.52 (1H, dd,= 7.8, 1.6 Hz, H-3), 7.02 (1H, brd,= 8.0 Hz, H-4), 6.94 (1H, t,= 7.4 Hz, H-5), 10.40 (1H, s, OH);13C-NMR (150 MHz, CDCl3): 175.0 (1-COOH), 162.3 (C-2), 137.0 (C-4), 131.1 (C-6), 119.7 (C-5), 111.8 (C-1), 118.0 (C-3)。經(jīng)數(shù)據(jù)比對[23],確定化合物14為水楊酸。
化合物15:淡黃色固體。ESI-MS/: 217 [M+Na]+,13C-NMR、APT譜確定分子式C10H10O4。1H-NMR (600 MHz, CDCl3): 7.69 (1H, d,= 16.2 Hz, H-7), 7.11 (1H, d,= 1.8 Hz, H-6), 6.99 (1H, dd,= 7.4, 1.2 Hz, H-2), 6.94 (1H, d,= 8.4 Hz, H-3), 6.31 (1H, d,= 15.6 Hz, H-8), 3.94 (3H, s, 5-OCH3);13C-NMR (150 MHz, CDCl3): 172.0 (C-9), 143.1 (C-7), 139.9 (C-5), 138.6 (C-4), 126.8 (C-1), 126.3 (C-2), 114.1 (C-3), 114.0 (C-6), 109.5 (C-8), 56.1 (5-OCH3)。經(jīng)數(shù)據(jù)比對[24],確定化合物15為反式-3-甲氧基-4-羥基肉桂酸。
化合物16:白色無定形粉末。ESI-MS/: 339 [M+Na]+,13C-NMR、APT譜確定分子式C20H28O3。1H-NMR (600 MHz, CDCl3): 7.22 (2H, s, H-11, 12), 7.16 (1H, s, H-14), 2.91 (1H, dd,= 16.0, 5.6 Hz, H-7b), 2.56 (1H, m, H-7a), 2.38 (1H, m, H-3b), 2.21 (1H, m, H-6b), 2.12 (1H, m, H-2b), 2.11 (1H, m, H-6a), 2.07 (1H, m, H-1b), 1.59 (1H, d,= 10.2 Hz, H-5), 1.58 (1H, m, H-2a), 1.56 (6H, s, H-16, 17), 1.33 (3H, s, H-18), 1.24 (1H, m, H-1a), 1.23 (1H, m, H-3a), 1.12 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 183.2 (C-19), 146.6 (C-9), 146.1 (C-13), 135.3 (C-8), 125.7 (C-11), 125.0 (C-14), 122.3 (C-12), 72.5 (C-15), 52.9 (C-5), 43.9 (C-4), 38.8 (C-1), 38.5 (C-10), 34.8 (C-3), 32.3 (C-7), 31.79 (C-16), 31.67 (C-17), 28.9 (C-18), 23.3 (C-20), 21.1 (C-6), 19.9 (C-2)。經(jīng)數(shù)據(jù)比對[20],確定化合物16為15-羥基松香酸。
化合物17:白色無定形粉末。ESI-MS/: 339 [M+Na]+,13C-NMR、APT譜確定分子式C20H28O3。1H-NMR (600 MHz, CDCl3): 7.19 (1H, d,= 7.2 Hz, H-11), 7.15 (1H, d,= 1.8 Hz, H-12), 7.13 (1H, d,= 1.8 Hz, H-14), 4.79 (1H, d,= 4.8 Hz, H-7), 2.33 (1H, m, H-1b), 2.29 (1H, d,= 7.2 Hz, H-5), 2.09 (1H, m, H-2b), 2.09 (1H, m, H-6b), 2.00 (1H, m, H-3b), 1.47 (1H, m, H-3a), 1.44 (1H, m, H-2a), 1.43 (1H, m, H-1a), 1.36 (1H, m, H-6a), 1.35 (6H, s, H-16, 17), 1.23 (1H, d,= 7.2 Hz, H-15), 1.10 (3H, s, H-19), 1.04 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 183.3 (C-18), 146.9 (C-9), 145.8 (C-13), 136.0 (C-8), 128.0 (C-14), 127.0 (C-12), 125.7 (C-11), 68.7 (C-7), 45.8 (C-5), 43.7 (C-4), 38.6 (C-1), 37.5 (C-3), 37.5 (C-10), 33.7 (C-15), 29.8 (C-6), 28.7 (C-20), 24.2 (C-17), 24.0 (C-16), 22.2 (C-19), 20.0 (C-2)。經(jīng)數(shù)據(jù)比對[25-26],確定化合物17為7β-羥基脫氫松香酸。
化合物18:白色無定形粉末。ESI-MS/: 339 [M+Na]+,13C-NMR、APT譜確定分子式C20H28O3。1H-NMR (600 MHz, CDCl3): 7.44 (1H, d,= 5.2 Hz, H-14), 7.17 (1H, d,= 7.8 Hz, H-11), 7.10 (1H, d,= 1.8 Hz, H-12), 4.75 (1H, dd,= 6.6, 6.0 Hz, H-6), 2.54 (1H, d,= 6.6 Hz, H-5), 2.33 (1H, m, H-1b), 2.18 (1H, m, H-7b), 2.03 (1H, m, H-2b), 1.98 (1H, m, H-3b), 1.96 (1H, m, H-2a), 1.59 (1H, d,= 12.2 Hz, H-3a), 1.59 (1H, d,= 12.6 Hz, H-7a), 1.33 (6H, s, H-16, 17), 1.25 (1H, m, H-1a), 1.22 (1H, m, H-15), 1.16 (3H, s, H-19), 0.60 (3H, s, H-20)。13C-NMR (150 MHz, CDCl3): 182.8 (C-18), 146.7 (C-9), 145.2 (C-13), 138.2 (C-8), 126.0 (C-14), 125.4 (C-12), 124.6 (C-11), 71.7 (C-6), 50.2 (C-5), 43.6 (C-4), 39.1 (C-10), 39.0 (C-1), 37.1 (C-3), 33.8 (C-15), 32.1 (C-7), 24.2 (C-17), 24.1 (C-16), 23.3 (C-20), 20.0 (C-2), 14.6 (C-19)。經(jīng)數(shù)據(jù)比對[25-26],確定化合物18為6-羥基脫氫松香酸。
化合物19:白色無定形粉末。ESI-MS/: 323 [M+Na]+,13C-NMR、APT譜確定分子式C19H24O3。1H-NMR (600 MHz, CDCl3): 7.65 (1H, d,= 8.2 Hz, H-12), 7.65 (1H, s, H-14), 7.24 (1H, d,= 5.4 Hz, H-11), 2.75 (1H, m, H-6b), 2.63 (1H, m, H-7b), 2.57 (3H, s, H-17), 2.47 (1H, m, H-7a), 2.44 (1H, m, H-1b), 2.18 (1H, m, H-2b), 2.07 (1H, m, H-3b), 1.98 (1H, m, H-6a), 1.54 (1H, d,= 11.6 Hz, H-5), 1.40 (1H, m, H-2a), 1.37 (3H, s, H-18), 1.37 (1H, m, H-1a), 1.17 (1H, m, H-3a), 1.01 (3H, s, H-20);13C-NMR (150 MHz, CDCl3): 198.4 (C-15), 183.0 (C-19), 153.8 (C-9), 136.0 (C-13), 134.6 (C-8), 129.0 (C-14), 126.1 (C-11), 126.0 (C-12), 52.6 (C-5), 44.0 (C-4), 39.3 (C-10), 38.5 (C-1), 36.9 (C-3), 32.1 (C-7), 29.9 (C-18), 24.2 (C-17), 22.1 (C-20), 20.8 (C-6), 20.3 (C-2)。經(jīng)數(shù)據(jù)比對[12],確定化合物19為角果酸G。
化合物20:白色無定形粉末。ESI-MS/: 339 [M+Na]+,13C-NMR、APT譜確定分子式C20H28O3。1H-NMR (600 MHz, CDCl3): 6.92 (1H, s, H-14), 6.91 (1H, d,= 7.6 Hz, H-12), 6.70 (1H, d,= 6.4 Hz, H-11), 2.88 (1H, m, H-7b), 2.82 (1H, m, H-15), 2.01 (1H, m, H-1b), 1.81 (1H, m, H-3b), 1.76 (1H, m, H-6b), 1.74 (1H, m, H-2b), 1.73 (1H, m, H-7a), 1.50 (3H, s, H-20), 1.49 (1H, m, H-3a), 1.44 (1H, m, H-2a), 1.43 (1H, m, H-1a), 1.34 (1H, m, H-6a), 1.26 (3H, s, H-19), 1.19 (6H, d,= 7.2 Hz, H-16, 17);13C-NMR (150 MHz, CDCl3): 180.8 (C-18), 151.5 (C-9), 141.5 (C-13), 128.1 (C-8), 128.1 (C-14), 125.1 (C-12), 115.2 (C-11), 85.3 (C-10), 55.4 (C-5), 48.8 (C-4), 36.8 (C-1), 36.6 (C-3), 33.5 (C-15), 29.2 (C-7), 26.5 (C-6), 24.4 (C-16, 17), 22.4 (C-20), 20.0 (C-2), 18.4 (C-19)。經(jīng)數(shù)據(jù)比對[7],確定化合物20為家地非諾酸 K。
由表1可知,部分二萜類化合物對RAW 264.7巨噬細胞具有抑制作用。以氫化可地松為陽性對照,化合物1、2、11和18具有微弱的抗炎活性,其中化合物11的抗炎活性相對較好,IC50值為0.256 mg/mL。具有抗炎活性的化合物1、2、11、18均為松香烷二萜結(jié)構(gòu),且六元環(huán)上的取代基都有1個羥基或1個羧基、或二者皆有,根據(jù)IC50值比較,這4個化合物的活性由強到弱依次為化合物11、18、2、1;根據(jù)結(jié)構(gòu)進行比較,化合物11和18六元環(huán)上的取代基都有1個羥基和1個羧基,化合物2六元環(huán)上的取代基有1個羥基,化合物1六元環(huán)的取代基是1個羧基。由此推斷,此類化合物六元環(huán)上的羧基或羥基可能是使化合物具有抗炎活性關(guān)鍵基團,并且羥基和羧基同時存在化合物的抗炎活性更好,取代基為羥基的化合物抗炎活性要優(yōu)于取代基為羧基的化合物。
表1 化合物對RAW 264.7細胞的抗炎作用
大苞荊芥是維吾爾族民醫(yī)習(xí)用藥材,具有鎮(zhèn)咳平喘、清熱利濕之功效,用于治療氣管炎、咳嗽氣喘、感冒發(fā)燒等癥。大苞荊芥常被作為神香草的替代品在民醫(yī)中廣泛用使用,因而對大苞荊芥的研究較少,缺乏對大苞荊芥化學(xué)成分及其藥用活性的系統(tǒng)研究。因此,本研究以大苞荊芥為研究對象,從其95%乙醇提取物石油醚和二氯甲烷萃取部位共分離純化了20種化合物,經(jīng)波譜解析和理化性質(zhì)鑒定,其中有18個化合物為二萜類化合物。NO作為炎癥組織損傷的重要致病因子,在急、慢性炎癥反應(yīng)的發(fā)生發(fā)展過程中發(fā)揮重要作用。當(dāng)免疫細胞遭受微生物內(nèi)毒素、炎癥介質(zhì)等刺激時,會生成大量的誘導(dǎo)型一氧化氮合成酶,產(chǎn)生NO進行免疫應(yīng)答[27]。因此,本研究通過各化合物對LPS誘導(dǎo)的小鼠RAW264.7巨噬細胞產(chǎn)生NO的影響來評價各化合物的抗炎活性。結(jié)果表明化合物1、2、11、18對LPS誘導(dǎo)的RAW264.7巨噬細胞NO的釋放均具有不同程度的抑制作用,其中化合物11的抑制作用相對較好,其IC50值為0.256 mg/mL。本研究闡明了大苞荊芥抗炎作用的藥效物質(zhì)基礎(chǔ),為大苞荊芥的進一步開發(fā)利用提供了科學(xué)依據(jù)。
利益沖突 所有作者均聲明不存在利益沖突
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Chemical constituents ofand its anti-inflammatory activities
ZHAO Peng-an1, 2, LIVARIDING Mahesuti2, MA Guo-xu2, 3, SUN Zhao-cui3, XU Xu-dong3, SHI Lei-ling2, GAO Xing-wang1
1. Xinjiang University, Urumqi 830046, China 2. Xinjiang Institute of Traditional Chinese Medicine and Ethnic Medicine, Urumqi 830002, China 3. Institute of Medicinal Plant Development, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100193, China
To elucidate the active substances inand their anti-inflammatory activities.The chemical constituents ofwere systematically isolated and purified by various chromatographic methods, and the structures of the compounds were identified by spectral analysis and physicochemical properties. Meanwhile, MTT colorimetry was used to test the anti-inflammatory activities of the compoundswith hydrocortisone as the positive control.Twenty compounds (two phenolic acids and 18 diterpenoids) were isolated and identified fromand identified as 7-oxodehydroabietic acid (1), dehydroabietol (2), lambertic acid (3), 3β,7α-dihydroxy-abieta-8,11,13-triene (4), triptobenzene L (5), 13-hydroxy-9-oxo-9,10-seco-abiet-8(14)-en-18,10a-olide (6), 8,11,13-abietatriene-7a,18-diol (7), 3β,19-dihydroxy-abieta-8,11,13- triene (8), 7α,15,18-trihydroxy-abieta-8,11,13-triene (9), abieta-8,11,13-trien-7β-ol (10), 6α-hydroxycallitrisic acid (11), 19-acetoxy-15-hydroxy-abieta-8,11,13-triene (12), 7a-hydroxy-abieta-8,11,13-trien-19-ylacetate (13), salicylis acid (14),-3-hydroxy-4-methoxy-cinnamic acid (15), 15-hydroxycallitrisic acid (16), 7β-hydroxycallitrisic acid (17), 6-hydroxycallitrisic acid (18), angustanoic acid G (19) and Jiadifenoic acid K (20). The results of anti-inflammatory activity showed that compound 11 inhibited the release of NO in LPS induced mouse macrophages RAW264.7, with IC50value of 0.256 mg/mL.Compounds 5, 6, 11 and 13 were isolated from Lamiaceae for the first time. Compounds 1, 3, 4, 7, 8, 10, and 12 are isolated fromgenus for the first time. Compounds 2, 9, 14 and 15 were isolated fromfor the first time. Compounds 11 had certain anti-inflammatory activity, which had the value for further in-depth research.
Benth.; diterpenoid; anti-inflammatory activity; triptobenzene L; 6α-hydroxycallitrisic acid; 7a-hydroxy- abieta-8,11,13-trien-19-ylacetate
R284.1
A
0253 - 2670(2023)22 - 7342 - 09
10.7501/j.issn.0253-2670.2023.22.011
2023-04-12
新疆維吾爾自治區(qū)公益性科研院所基本科研業(yè)務(wù)經(jīng)費資助項(Ky2023zmy005);中國醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程(2022-I2M-1-017)
趙鵬安(1998—),男,碩士研究生,研究方向為中藥學(xué)。E-mail: 1049867556@qq.com
通信作者:石磊嶺(1977—),男,碩士,研究員,研究方向為中藥學(xué)研究。E-mail: shileiling@sina.com
高興旺(1983—),男,博士,主要從事天然產(chǎn)物化學(xué)研究。E-mail: gxw@xju.edu.cn
[責(zé)任編輯 王文倩]