从鸡血藤中获得醇提物(alcohol extract,ESs)及水提物(water extracts,WSs),研究其对α-淀粉酶和α-葡萄糖苷酶的抑制作用。结果表明,ESs的抑制效果显著高于WSs。为深入研究鸡血藤对2种酶的抑制作用,采用不同极性溶剂对ESs进一步萃取,获得W-ESs、D-ESs、N-ESs、E-ESs四个组分。实验结果表明,在同一浓度范围,ESs及4个组分的抑制效果存在差异。对α-淀粉酶的抑制效果为:W-ESs>D-ESs>ESs>N-ESs>E-ESs;对α-葡萄糖苷酶的抑制效果为:W-ESs>ESs>N-ESs>E-ESs>D-ESs。其中W-ESs对2种酶的抑制作用最好,IC50值分别为(0.88±0.02) mg/mL和(15.82±2.79) μg/mL。绘制Lineweaver-Burk 曲线确定酶抑制类型,结果显示,WSs、ESs、W-ESs及N-ESs对α-淀粉酶为反竞争性抑制,D-ESs和E-ESs为竞争性和非竞争性混合抑制。对于α-葡萄糖苷酶,W-ESs、D-ESs和E-ESs为反竞争性抑制类型,而WSs、ESs和N-ESs为反竞争性和非竞争性混合的抑制类型。研究结果表明,鸡血藤具有开发为降血糖药物的潜力。
In this study, ethanol extracts (ESs) and water extracts (WSs) were obtained from Spatholobus suberectus. The inhibition of α-amylase and α-glucosidase were determined. The results showed that the inhibition effect of ESs was significantly higher than that of WSs. To further explore the inhibition of the two enzymes by S. suberectus, the ESs was further separated by different polar solvents and obtained four components: W-ESs, D-ESs, E-ESs, and N-ESs. The different inhibitory effects of ESs and the four components at the same concentration range were as follows, α-amylase: W-ESs>D-ESs>ESs>N-ESs>E-ESs; α-glucosidase: W-ESs>ESs>N-ESs>E-ESs>D-ESs. W-ESs showed the best inhibition effect for both enzymes, with IC50 values of (0.88±0.02) mg/mL and (15.82±2.79) μg/mL. The Lineweaver-Burk curve was plotted to determine the type of enzyme inhibition. The results showed that WSs, ESs, W-ESs, and N-ESs were sensitive to α-amylase, which were the type of anti-competitive inhibition. D-ESs and E-ESs were competitive and non-competitive mixed inhibition. For α-glucosidases, WEs, D-ESs and E-ESs were the type of anti-competitive inhibition, while WSs, ESs and N-ESs were competitive and non-competitive mixed inhibition type. Our results showed that S. suberectus had the potential to be developed as a hypoglycemic drug.
[1] TU W J, XUE Y M, NIE D.The prevalence and treatment of diabetes in China from 2013 to 2018[J].JAMA, 2022, 327(17):1706.
[2] HIYOSHI T, FUJIWARA M, YAO Z M.Postprandial hyperglycemia and postprandial hypertriglyceridemia in type 2 diabetes[J].Journal of Biomedical Research, 2017, 33(1):1-16.
[3] JAGADEESAN G, MUNIYANDI K, MANOHARAN A L, et al.Understanding the bioaccessibility, α-amylase and α-glucosidase enzyme inhibition kinetics of Allmania nodiflora (L.) R.Br.ex Wight polyphenols during in vitro simulated digestion[J].Food Chemistry, 2022, 372:131294.
[4] 顾觉奋, 陈紫娟.α-葡萄糖苷酶抑制剂的研究及应用[J].药学进展, 2009, 33(2):62-67.
GU J F, CHEN Z J.The studies and applications of α-glucosidase inhibitors[J].Progress in Pharmaceutical Sciences, 2009, 33(2):62-67.
[5] 李洪梅. α-葡萄糖苷酶抑制剂的临床应用[J].中国医刊, 2007, 42(10):19-21.
LI H M.Clinical application of α-glucosidase inhibitors[J].Chinese Journal of Medicine, 2007, 42(10):19-21.
[6] 国家药典委员会. 中华人民共和国药典-二部:2010年版[M].北京:中国医药科技出版社, 2010.
Chinese Pharmacopoeia Commission. Chinese pharmacopoeia-Part Ⅱ: 2010 edition[M]. Beijing: The Medicine Science and Technology Press of China, 2010.
[7] 刘静, 王晓静, 戴忠, 等.鸡血藤研究进展[J].中国药事, 2019, 33(2):188-194.
LIU J, WANG X J, DAI Z, et al.Research progress of Spatholobi caulis[J].Chinese Pharmaceutical Affairs, 2019, 33(2):188-194.
[8] 苏贤君. 中药鸡血藤Spatholobus suberectus Dunn.化学成分的研究[D].广州:广州中医药大学, 2016.
SU X J.Research on the chemical constituents of Spatholobus suberectus Dunn.[D].Guangzhou:Guangzhou University of Chinese Medicine, 2016.
[9] 唐旗羚, 曹颖男, 徐娟娟, 等.鸡血藤组方提取物的抗肿瘤作用及其机制初探[J].中国医药指南, 2018, 16(15):34;39.
TANG Q L, CAO Y N, XU J J, et al.Study of anti-cancer effect and the mechanism of Millettia formulation extract[J].Guide of China Medicine, 2018, 16(15):34;39.
[10] FU Y F, JIANG L H, ZHAO W D, et al.Immunomodulatory and antioxidant effects of total flavonoids of Spatholobus suberectus Dunn on PCV2 infected mice[J].Scientific Reports, 2017, 7:8676.
[11] HOU X K, MAO J S.Long noncoding RNA SNHG14 promotes osteosarcoma progression via miR-433-3p/FBXO22 axis[J].Biochemical and Biophysical Research Communications, 2020, 523(3):766-772.
[12] 李丽, 王林萍.鸡血藤总黄酮对大鼠急性心肌缺血的保护作用[J].中成药, 2015, 37(10):2 303-2 306.
LI L, WANG L P.Protective effect of total flavonoids from Spatholobus suberectus on acute myocardial ischemia in rats[J].Chinese Traditional Patent Medicine, 2015, 37(10):2 303-2 306.
[13] ZHANG C H, LIU J Y, HE X Y, et al.Caulis spatholobi ameliorates obesity through activating brown adipose tissue and modulating the composition of gut microbiota[J].International Journal of Molecular Sciences, 2019, 20(20):5150.
[14] 袁莎莎, 杨宏杰, 范朝华, 等.鸡血藤总黄酮对2型糖尿病模型大鼠血脂、血液流变学的影响[J].中医学报, 2020, 35(10):2 178-2 181.
YUAN S S, YANG H J, FAN C H, et al.Effects of Spatholobus spatholobi total flavonoids on blood lipids and hemorheology in type 2 diabetic rat[J].Acta Chinese Medicine, 2020, 35(10):2 178-2 181.
[15] ZHAO P J, ALAM M B, LEE S H, et al.Spatholobus suberectus exhibits antidiabetic activity in vitro and in vivo through activation of AKT-AMPK pathway[J].Evidence-Based Complementary and Alternative Medicine:ECAM,2017, 2017:6091923.
[16] 王宁丽, 魏鉴腾, 祝潇, 等.基于碘-淀粉显色的α-淀粉酶抑制剂筛选方法研究及应用[J].中国海洋药物, 2017, 36(5):75-78.
WANG N L, WEI J T, ZHU X, et al.Study and application of the method for screening of α-amylase inhibitors based on iodine-starch colorimetry[J].Chinese Journal of Marine Drugs, 2017, 36(5):75-78.
[17] JIANG L L, WANG Z, WANG X Y, et al.Exploring the inhibitory mechanism of piceatannol on α-glucosidase relevant to diabetes mellitus[J].RSC Advances, 2020, 10(8):4 529-4 537.
[18] KIM J H, LEE S Y, PARK J M, et al.Antioxidant activity and inhibition activity against α-amylase and α-glucosidase of Juniperus rigida Sieb extracts[J].Korean Journal of Food Preservation, 2014, 21(3):396-403.
[19] 庄远杯, 凌梅娣, 魏爱红, 等.石韦不同极性萃取物体外降血糖活性研究[J].广西植物, 2022, 42(5):855-859.
ZHUANG Y B, LING M D, WEI A H, et al.Hypoglycemic activities of different solvent extracts from Pyrrosia Lingua in vitro[J].Guihaia, 2022, 42(5):855-859.
[20] 温正辉, 凌梅娣, 余思萍, 等.蒲桃不同药用部位乙醇提取物对α-葡萄糖苷酶和α-淀粉酶活性的抑制作用研究[J].中国药房, 2019, 30(23):3 246-3 251.
WEN Z H, LING M D, YU S P, et al.Study on inhibitory effects of ethanol extract of different medicinal parts from Syzygium jambos on the activities of α-glycosidase and α-amylase[J].China Pharmacy, 2019, 30(23):3 246-3 251.
[21] 许建和. 生物催化工程[M].上海:华东理工大学出版社, 2008.
XU J H. Biocatalysis Engineering[M]. Shanghai: East China University of Science and Technology Press, 2008.
[22] 郑丽婷, 周鸿, 刘奕明, 等.黄柏碱对α-葡萄糖苷酶的体外抑制作用[J].南京中医药大学学报, 2020, 36(6):853-858.
ZHENG L T, ZHOU H, LIU Y M, et al.Inhibitory effect of phellodendrine on α-glucosidase in vitro[J].Journal of Nanjing University of Traditional Chinese Medicine, 2020, 36(6):853-858.
[23] 包正宇, 樊铭聪, 李言, 等.乌饭树树叶蓝黑色素对α-葡萄糖苷酶的抑制活性研究[J].食品与机械, 2021, 37(7):165-170;176.
BAO Z Y, FAN M C, LI Y, et al.Study on the α-glucosidase inhibition of the dark blue pigment from the leaves of Vaccinium bracteatum Thunb[J].Food & Machinery, 2021, 37(7):165-170;176.
[24] 李美萍, 侯健, 刘燕, 等.细叶韭花醇提物体外抗氧化及对α-葡萄糖苷酶抑制作用的研究[J].中国调味品, 2022, 47(2):30-34.
LI M P, HOU J, LIU Y, et al.Study on antioxidation of ethanol extract from Allium tenuissimum L.flowers in vitro and its inhibition on α-glucosidase[J].China Condiment, 2022, 47(2):30-34.
