中药砂仁降血糖活性成分研究

	中药砂仁降血糖活性成分研究
	丁敏
导师	耿长安
关键词	砂仁，姜科，降木脂素，降血糖活性，促GLP-1分泌，GPa抑制活性 Amomum villosum Zingiberaceae norlignans antidiabetic effects GLP-1 stimulation GPa inhibition
摘要	糖尿病 (diabetes mellitus) 是由于胰岛素分泌不足或其生物效应受损而引起人体糖、蛋白质、脂肪和电解质代谢紊乱的疾病。现有治疗药物主要包括胰岛素和口服降糖药，然而长期使用敏感性降低和易引发低血糖等，不能满足临床需要，亟需发现新型降糖药物。中药砂仁 (Amomi Fructus) 为姜科豆蔻属植物阳春砂、绿壳砂或海南砂的干燥成熟果实，具有抗菌、抗炎、胃肠保护和降血糖等活性。我们发现，砂仁的乙醇提取物在50.0 μg/mL时，对GPa酶的抑制率为79.1%，同时在STC-1细胞上具有促胰高血糖素样肽-1 (GLP-1) 分泌的活性，在50.0 μg/mL时，促泌率为100.1%。为了揭示砂仁的降血糖活性成分，本研究以α-葡萄糖苷酶和GPa酶抑制活性为导向，利用硅胶、葡聚糖凝胶LH-20、Rp-C18、MCI gel CHP 20P、制备TLC、半制备HPLC等色谱分离手段，从中分离鉴定43个化合物，其结构通过HRESIMS、1D和2D NMR、IR和UV等波谱技术以及ECD计算方法鉴定。化合物结构类型涉及9个木脂素 (1~7, 25, 26)，6个单萜 (8~13)，2个桉烷型倍半萜 (14, 15)，2个半日花烷型二萜 (16, 17)，6个达玛烷型三萜苷 (18~23)，1个二芳基庚烷 (24)，2个黄酮 (27, 28)，2个苯丙素 (31, 32) 以及3个苯乙酮 (33~35)和10个其他类型化合物 (29, 30, 36~43)。8个新化合物分别鉴定为：(?) 砂仁素A (1)、(+) 砂仁素A (2)、(?) 砂仁素B (3)、(+) 砂仁素B (4)、(?) 砂仁素C (5)、(+) 砂仁素C (6)、砂仁素D (7)和kravanhin A 3-O-β-D-glucopyranoside (8)，其中化合物1/2、3/4和5/6互为对映异构体。采用三种酶学 (α-葡萄糖苷酶、PTP1B和GPa) 和STC-1细胞模型对砂仁中分离得到的化合物进行活性评价。结果表明，砂仁素A (1/2) 具有显著的促GLP-1分泌活性，在25.0和12.5 μM 时促泌率分别为375.1%和166.9%，明显高于阳性对照OEA (163.1%, 25.0 μM)，砂仁素B (3/4)在25.0和12.5 μM 时，对GLP-1的促泌率分别为222.7%和62.7%。4个莰烷型单萜 (8~11) 和1个半日花烷型二萜 (17) 在25.0 μM浓度时，对GLP-1的促泌率分别为94.6 %、258.4%、398.6%、483.3%和692.8%。萜类化合物 (1R,2S,4R,7S)-vicodiol 9-O-β-D-glucopyranoside (9) 和三七皂苷 R1 (18) 对GPa酶具有明显的抑制活性，IC50分别为78.6和98.7 μM；酚性成分异槲皮苷 (28) 和3-甲氧基-1, 2苯二酚 (37) 对PTP1B具有较好的抑制活性，IC50分别为41.9 μM和147.5 μM。本研究初步揭示了中药砂仁的降血糖活性成分，为砂仁的降血糖应用提供了化学和药理学依据。本文第二章对中药砂仁的化学成分和药理活性研究进展进行综述，总结了近二十年从砂仁中分离得到的98个化学成分，化合物类型主要包括单萜、倍半萜、二萜、二芳基庚烷和黄酮等；药理活性主要涉及胃肠保护、降血糖、抗炎、镇痛、抗菌和抗氧化等。; Diabetes mellitus is a disorder of glucose, protein, fat or electrolyte metabolism in the body due to insufficient insulin secretion and/or insulin resistance in organs. Exogenous insulins and oral hypoglycaemic drugs are the main treatment measures for diabetes mellitus, whereas their efficacies are unsatisfactory due to undesirable hypoglycemia, side effects and drug resistance. The dried fruit of Amomum villosum (Amomi Fructus) is an important spices and traditional Chinese medicine in Zingiberaceae family with antibacterial, anti-inflammatory, gastrointestinal protection and hypoglycemic activities. In this study, the EtOH extract of Amomi Fructus was revealed with GPa inhibitory activity with an inhibitory ratio of 79.1% at 50.0 μg/mL. In addition, the EtOH extract also showed glucagon-like peptide-1 (GLP-1) stimulation on STC-1 cells by 100.1%, at the concentration of 50.0 μg/mL. In order to clarify the antidiabetic constituents, 43 compounds were isolated by various chromatographic methods including silica gel, sephadex LH-20, Rp-C18, MCI gel CHP 20P column chromatography, preparative thin layer chromatography and HPLC. Their structures were determined by extensive spectroscopic data (1D and 2D NMR, HRESIMS, IR, UV and [α]D) and ECD calculations. The isolated compounds were identified as nine lignans (1~7, 25, 26), six monoterpenes (8~13), two sesquiterpenes (14, 15), two diterpenes (16, 17), two triterpenes (18~23), one diarylheptanoid (24), two flavonoids (27, 28), two phenylpropanoid (31, 32), three acetophenone (33~35), and ten other types compounds. Eight new compounds were characterized as (?) amovillosumin A (1), (+) amovillosumin A (2), (?) amovillosumin B (3), (+) amovillosumin B (4), (?) amovillosumin C (5), (+) amovillosumin C (6), amovillosumin D (7) and kravanhins A 3-O-β-D-glucopyranoside (8), of which compounds 1/2, 3/4 and 5/6 were three pairs of enantiomers. The antidiabetic effects of the isolates were analyzed by three enzymes and STC-1 cells. Amovillosumin A (1/2) showed obvious activity in stimulating GLP-1 secretion by 166.9 % and 375.1 % at 12.5 and 25.0 μM, more potent than the positive control, oleoylethanolamide (OEA, 163.1%, 25.0 μM). Amovillosumin B (2) obviously increased the GLP-1 secretion by 62.7% (12.5 μM) and 222.7% (25.0 μM), comparable to the positive control. Four monoterpenoids (8~11) and one labdane diterpenoid (17) showed GLP-1 stimulation at 25.0 μM with promotion rates of 94.6 %, 258.4%, 398.6%, 483.3% and 692.8%, more potent than OEA (133.2%, 25.0 μM). Two terpenoids (1R,2S,4R,7S)-vicodiol 9-O-β-D-glucopyranoside (9) and notoginsenoside R1 (18) showed GPa inhibition with IC50 values of 78.6 and 98.7 μM. Compounds 28 and 37 showed obvious PTP1B inhibitory activity with IC50 values of 41.9 μM and 147.5 μM. The second chapter reviewed the research progress of the chemical constituents and pharmacological activities of A. villosum. A total of 98 chemical constituents were isolated from A. villosum.
语种	中文
	2022-05
学位授予单位	中国科学院大学
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/75121
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	丁敏. 中药砂仁降血糖活性成分研究[D]. 中国科学院大学,2022.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
丁敏-中药砂仁降血糖活性成分研究--终稿（6349KB）	学位论文		限制开放	CC BY-NC-SA	请求全文