Article(id=1322925863938957621, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322925862630339452, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.07.001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793339704736, onlineDateStr=2026-10-30, pubDate=1752076800000, pubDateStr=2025-07-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339704736, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339704736, creator=13701087609, updateTime=1793339704736, updator=13701087609, issue=Issue{id=1322925862630339452, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='7', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1752076800000', pubDateStr='2025-07-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339704424, creator='13701087609', updateTime=1793340532184, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322929334587105328, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322925862630339452, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322929334587105329, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322925862630339452, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1, endPage=5, ext={EN=ArticleExt(id=1322925864719098166, articleId=1322925863938957621, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Exploration on Hypoglycemic Mechanism of Yuzhu(Polygonatum odoratumin)Saponin Based on Sweet Taste and Sweet Receptors, columnId=1256183423710970242, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=New Opinion of Ph.D. Supervisor, runingTitle=null, highlight=null, articleAbstract=
Objective

To define“sweetness”in traditional Chinese medicine based on sweet taste receptors and explore the hypoglycemic mechanism of Yuzhu(Polygonatum odoratumin)saponin.

Methods

The diabetic model was established by streptozocin(STZ)method. After successfully modeling,the mice were randomly divided into model group,metformin group,low dose of Yuzhu(Polygonatum odoratumin)saponin group,high dose of Yuzhu(Polygonatum odoratumin)saponin group,low dose of Yuzhu(Polygonatum odoratumin)saponin+Lac(Lactisole)group and high dose of Yuzhu(Polygonatum odoratumin)saponin+Lac group. The mice without modeling were taken as the normal group. Fasting blood glucose and glucose tolerance were tested 7 weeks after administration of the drug. After 8 weeks of drug administration,the levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were determined by biochemical kits. The morphology of pancreas and liver tissues were observed by hematoxylin-eosin staining(HE),and the contents of glucagon-like peptide-1(GLP-1)and insulin were determined by enzyme linked immunosorbent assay(ELISA). HuTu-80 was used as a cellular model,and the contents of GLP-1 and Cyclic adenosine monophosphate(cAMP)were detected by ELISA. The intensity of Ca2+fluorescence was detected by laser confocal.

Results

Yuzhu(Polygonatum odoratumin)saponin significantly reduced the levels of fasting blood glucose,TC,TG and LDL-C in diabetic mice(P<0.05). Yuzhu(Polygonatum odoratumin)saponin increased the levels of HDL-C,GLP-1 and insulin(P<0.05),and improved the morphology of the pancreas and liver tissues. Yuzhu(Polygonatum odoratumin)saponin significantly increased the contents of cAMP,Ca2+and GLP-1 after their action on cells(P<0.05). However,the action of Yuzhu(Polygonatum odoratumin)saponin could be significantly inhibited by the sweet taste receptor inhibitor Lactisole(P<0.05).

Conclusion

Yuzhu(Polygonatum odoratumin)saponin agitates sweet taste receptors due to the sweet taste,which in turn promotes GLP-1 secretion and exerts hypoglycemic effects.

, authors=Huasheng YANG, Yin XIAO, Ting LI, Yawen DUAN, Lu WU, authorsList=Huasheng YANG, Yin XIAO, Ting LI, Yawen DUAN, Lu WU, authorCompany=null, correspAuthors=Lu WU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1322925869391552847, articleId=1322925863938957621, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=基于甘味及甜味受体探讨玉竹皂苷降血糖作用机制, columnId=1256183426563096985, journalTitle=中华中医药学刊, columnName=博士导师新论, runingTitle=null, highlight=null, articleAbstract=
目的

基于甜味受体界定中药“甘味”,并以此探讨玉竹皂苷降血糖的作用机制。

方法

采用链脲佐菌素制备糖尿病模型,造模成功后随机分为模型组、二甲双胍组、玉竹皂苷低剂量组、玉竹皂苷高剂量组、玉竹皂苷低剂量+2-(4-甲氧基苯氧基)丙酸(Lactisole,Lac)组、玉竹皂苷高剂量+Lac组,另取未造模小鼠为正常组。给药7周后,检测空腹血糖及糖耐量;给药8周后,采用生化试剂盒测定总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)含量,苏木精-伊红染色法(hematoxylin-eosin staining,HE)观察胰腺及肝组织形态,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)、胰岛素的含量;以HuTu-80为细胞模型,ELISA检测GLP-1、环磷酸腺苷(cyclic adenosine monophosphate,cAMP)的含量,激光共聚焦检测Ca2+荧光强度。

结果

玉竹皂苷能显著降低糖尿病小鼠空腹血糖及TC、TG、LDL-C水平(P<0.05),升高HDL-C、GLP-1、胰岛素水平(P<0.05),改善胰腺及肝组织形态;玉竹皂苷作用于HuTu-80细胞后,可显著增加cAMP、Ca2+、GLP-1的含量(P<0.05)。然而,玉竹皂苷的作用能被甜味受体抑制剂Lactisole明显抑制(P<0.05)。

结论

玉竹皂苷因味甘而激动甜味受体,进而促进GLP-1分泌,发挥降血糖作用。

, authors=杨华生, 肖音, 李婷, 段亚文, 吴璐, authorsList=杨华生, 肖音, 李婷, 段亚文, 吴璐, authorCompany=null, correspAuthors=吴璐, authorNote=

杨华生(1974-),男,湖南衡阳人,教授,博士研究生导师,博士,研究方向:传统中医药理论及中药制剂。

, correspAuthorsNote=
吴璐(1980-),女,湖南长沙人,副教授,硕士,研究方向:传统中医药理论及中药制剂。E-mail:。
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杨华生(1974-),男,湖南衡阳人,教授,博士研究生导师,博士,研究方向:传统中医药理论及中药制剂。

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杨华生(1974-),男,湖南衡阳人,教授,博士研究生导师,博士,研究方向:传统中医药理论及中药制剂。

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组别nTCTGLDL-CHDL-C
NC组62.05±0.301.12±0.220.31±0.081.48±0.24
DM组64.51±0.67*2.10±0.37*0.48±0.06*0.93±0.14*
PC组62.86±0.28▲1.26±0.28▲0.34±0.06▲1.41±0.20▲
LD组63.36±0.56▲1.55±0.31▲0.42±0.061.31±0.23▲
HD组63.10±0.71▲1.40±0.21▲0.37±0.04▲1.38±0.29▲
LD+Lac组64.14±0.46#1.95±0.35#0.47±0.050.98±0.20#
HD+Lac组63.88±0.65Δ1.79±0.36Δ0.43±0.051.09±0.09Δ
), ArticleFig(id=1322925874365997439, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925863938957621, language=CN, label=表1, caption=

玉竹皂苷对小鼠血清中血脂水平的影响(±s)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别nTCTGLDL-CHDL-C
NC组62.05±0.301.12±0.220.31±0.081.48±0.24
DM组64.51±0.67*2.10±0.37*0.48±0.06*0.93±0.14*
PC组62.86±0.28▲1.26±0.28▲0.34±0.06▲1.41±0.20▲
LD组63.36±0.56▲1.55±0.31▲0.42±0.061.31±0.23▲
HD组63.10±0.71▲1.40±0.21▲0.37±0.04▲1.38±0.29▲
LD+Lac组64.14±0.46#1.95±0.35#0.47±0.050.98±0.20#
HD+Lac组63.88±0.65Δ1.79±0.36Δ0.43±0.051.09±0.09Δ
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基于甘味及甜味受体探讨玉竹皂苷降血糖作用机制
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杨华生 , 肖音 , 李婷 , 段亚文 , 吴璐
中华中医药学刊 | 博士导师新论 2025,43(7): 1-5
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中华中医药学刊 |博士导师新论 2025 , 43 (7) : 1 -5
基于甘味及甜味受体探讨玉竹皂苷降血糖作用机制
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[Author(id=1322925869659988308, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925863938957621, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1322925869735485782, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925863938957621, authorId=1322925869659988308, language=EN, stringName=Huasheng YANG, firstName=Huasheng, middleName=null, lastName=YANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=Jiangxi University of Chinese Medicine,Nanchang 330004,Jiangxi,China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1322925869806788951, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322925863938957621, authorId=1322925869659988308, language=CN, stringName=杨华生, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=江西中医药大学,江西 南昌 330004, bio={"content":"

杨华生(1974-),男,湖南衡阳人,教授,博士研究生导师,博士,研究方向:传统中医药理论及中药制剂。

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杨华生(1974-),男,湖南衡阳人,教授,博士研究生导师,博士,研究方向:传统中医药理论及中药制剂。

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杨华生, 肖音, 李婷, 段亚文, 吴璐
作者信息
  • 江西中医药大学,江西 南昌 330004
通讯作者:
吴璐(1980-),女,湖南长沙人,副教授,硕士,研究方向:传统中医药理论及中药制剂。E-mail:。
作者简介:

杨华生(1974-),男,湖南衡阳人,教授,博士研究生导师,博士,研究方向:传统中医药理论及中药制剂。

Exploration on Hypoglycemic Mechanism of Yuzhu(Polygonatum odoratumin)Saponin Based on Sweet Taste and Sweet Receptors
Huasheng YANG, Yin XIAO, Ting LI, Yawen DUAN, Lu WU
Affiliations
  • Jiangxi University of Chinese Medicine,Nanchang 330004,Jiangxi,China
出版时间: 2025-07-10 doi: 10.13193/j.issn.1673-7717.2025.07.001
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目的

基于甜味受体界定中药“甘味”,并以此探讨玉竹皂苷降血糖的作用机制。

方法

采用链脲佐菌素制备糖尿病模型,造模成功后随机分为模型组、二甲双胍组、玉竹皂苷低剂量组、玉竹皂苷高剂量组、玉竹皂苷低剂量+2-(4-甲氧基苯氧基)丙酸(Lactisole,Lac)组、玉竹皂苷高剂量+Lac组,另取未造模小鼠为正常组。给药7周后,检测空腹血糖及糖耐量;给药8周后,采用生化试剂盒测定总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)含量,苏木精-伊红染色法(hematoxylin-eosin staining,HE)观察胰腺及肝组织形态,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)、胰岛素的含量;以HuTu-80为细胞模型,ELISA检测GLP-1、环磷酸腺苷(cyclic adenosine monophosphate,cAMP)的含量,激光共聚焦检测Ca2+荧光强度。

结果

玉竹皂苷能显著降低糖尿病小鼠空腹血糖及TC、TG、LDL-C水平(P<0.05),升高HDL-C、GLP-1、胰岛素水平(P<0.05),改善胰腺及肝组织形态;玉竹皂苷作用于HuTu-80细胞后,可显著增加cAMP、Ca2+、GLP-1的含量(P<0.05)。然而,玉竹皂苷的作用能被甜味受体抑制剂Lactisole明显抑制(P<0.05)。

结论

玉竹皂苷因味甘而激动甜味受体,进而促进GLP-1分泌,发挥降血糖作用。

甘味  /  甜味受体  /  玉竹  /  胰高血糖素样肽-1
Objective

To define“sweetness”in traditional Chinese medicine based on sweet taste receptors and explore the hypoglycemic mechanism of Yuzhu(Polygonatum odoratumin)saponin.

Methods

The diabetic model was established by streptozocin(STZ)method. After successfully modeling,the mice were randomly divided into model group,metformin group,low dose of Yuzhu(Polygonatum odoratumin)saponin group,high dose of Yuzhu(Polygonatum odoratumin)saponin group,low dose of Yuzhu(Polygonatum odoratumin)saponin+Lac(Lactisole)group and high dose of Yuzhu(Polygonatum odoratumin)saponin+Lac group. The mice without modeling were taken as the normal group. Fasting blood glucose and glucose tolerance were tested 7 weeks after administration of the drug. After 8 weeks of drug administration,the levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were determined by biochemical kits. The morphology of pancreas and liver tissues were observed by hematoxylin-eosin staining(HE),and the contents of glucagon-like peptide-1(GLP-1)and insulin were determined by enzyme linked immunosorbent assay(ELISA). HuTu-80 was used as a cellular model,and the contents of GLP-1 and Cyclic adenosine monophosphate(cAMP)were detected by ELISA. The intensity of Ca2+fluorescence was detected by laser confocal.

Results

Yuzhu(Polygonatum odoratumin)saponin significantly reduced the levels of fasting blood glucose,TC,TG and LDL-C in diabetic mice(P<0.05). Yuzhu(Polygonatum odoratumin)saponin increased the levels of HDL-C,GLP-1 and insulin(P<0.05),and improved the morphology of the pancreas and liver tissues. Yuzhu(Polygonatum odoratumin)saponin significantly increased the contents of cAMP,Ca2+and GLP-1 after their action on cells(P<0.05). However,the action of Yuzhu(Polygonatum odoratumin)saponin could be significantly inhibited by the sweet taste receptor inhibitor Lactisole(P<0.05).

Conclusion

Yuzhu(Polygonatum odoratumin)saponin agitates sweet taste receptors due to the sweet taste,which in turn promotes GLP-1 secretion and exerts hypoglycemic effects.

sweet taste  /  sweet receptor  /  Yuzhu(Polygonatum odoratumin)  /  glucagon-like peptide-1
杨华生, 肖音, 李婷, 段亚文, 吴璐. 基于甘味及甜味受体探讨玉竹皂苷降血糖作用机制. 中华中医药学刊, 2025 , 43 (7) : 1 -5 . DOI: 10.13193/j.issn.1673-7717.2025.07.001
Huasheng YANG, Yin XIAO, Ting LI, Yawen DUAN, Lu WU. Exploration on Hypoglycemic Mechanism of Yuzhu(Polygonatum odoratumin)Saponin Based on Sweet Taste and Sweet Receptors[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (7) : 1 -5 . DOI: 10.13193/j.issn.1673-7717.2025.07.001
甘味,又称甜味,是中药五味之一。补虚类中药多为甘味,《中华人民共和国药典》收载的甘味药达238种,占比38.4%[1]。中药味甘有两个层面的含义,或滋味是“甘”,或具有“甘补/缓/和”的效应。因此,中药的甘可能是基于“滋味”,也可能是基于“效味”。这种二元的界定甘味的方法,可能对中药的“味-效”研究产生一定的困扰;同时,有的中药不仅滋味甘,其效也补/缓/和,那么这类中药甘味的界定标准就难以确定。现代研究表明,“甘”的产生与味觉感受器有关,而这种感受器即为“甜味受体”[2]。
甜味受体属于味觉受体第一家族,其分布广泛,在舌、肠道等均有表达。激活甜味受体,可产生甜味或相应效应[3]。“物质”激活舌头味蕾细胞中甜味受体,启动甜味信号,产生并传递甜味;基于滋味,这种物质可界定为“甘味”。“物质”激活肠道甜味受体,α-味导素活化,细胞内环磷酸腺苷(cyclic adenosine monophosphate,cAMP)浓度上升,Ca2+内流增加,启动甜味信号,胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)分泌增加,产生相应效应[4-7];基于效味,这种物质可界定为“甘味”。可见,甜味受体与中药的滋味、效味均有关联。因此可从甜味受体的角度,初步对中药“甘味”做一新的界定,即“凡可激动甜味受体,启动甜味信号的物质,均为甘味”。同时,有的物质虽不激动甜味受体,但可促进甜味受体表达而增强甜味信号强度,在葡萄糖等内源性甜味物质的介导下,也能促进GLP-1的分泌,这些物质也可界定为“甘味”,如甘味药含有的多糖。基于以上分析,本文提出“凡可启动、增强甜味信号的物质,均为甘味”。
玉竹为百合科植物玉竹Polygonatum odoratum(Mill.)Druce的干燥根茎,功能“养阴润燥,生津止渴”,中医临床上常用于治疗胃阴虚、肺阴虚证[8]。玉竹主要含有多糖、皂苷、黄酮等成分,现代常用于糖尿病的治疗[9-10]。玉竹味“甘”,基于前述“凡可启动、增强甜味信号的物质,均为甘味”,可以推测,玉竹中应含有可启动、增强甜味信号的物质。本项目组前期研究也发现,玉竹中的皂苷可促进阴虚动物GLP-1的分泌[11]。因此玉竹皂苷可能是玉竹降血糖的物质基础之一,其机制可能与其启动、增强甜味信号有关。故本研究先从甜味受体的角度,构建界定中药“甘味”的新方法;采用链脲佐菌素(streptozotocin,STZ)制备糖尿病模型,研究玉竹皂苷对糖尿病小鼠空腹血糖、糖耐量、血脂、GLP-1、胰岛素、胰脏和肝脏组织形态的影响,并与甜味受体抑制剂2-(4-甲氧基苯氧基)丙酸(Lactisole,Lac)进行比较研究;采用细胞模型研究玉竹皂苷对甜味信号通路上信号分子cAMP、Ca2+的影响,进一步验证玉竹皂苷对甜味受体的激动作用。
KM小鼠,清洁级,体质量20~22 g,由江西中医药大学实验动物科技中心提供,许可证号:SCXK(赣)2018-0003。本研究所有实验操作均符合实验动物使用指导原则,方案取得江西中医药大学伦理委员会审核批准(批准号:JZSYDWLL-20201215)。
HuTu-80人十二指肠腺癌细胞(上海富衡生物科技有限公司,批号:FH0536)。
玉竹药材(安徽道源堂中药饮片有限公司,批号:200901);盐酸二甲双胍片(中美上海施贵宝制药有限公司,批号:AbS0267)。链脲佐菌素(北京索莱宝科技有限公司,批号:506J022);总胆固醇(total cholesterol,TC)试剂盒、甘油三酯(triglyceride,TG)试剂盒、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)试剂盒、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)试剂盒(南京建成生物工程研究所,批号:20220922、20220921、20220922、20220923);小鼠GLP-1试剂盒(上海恒远生物科技有限公司,批号:H2106Y28);GLP-1酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)试剂盒(武汉菲恩生物科技有限公司,批号:H1053G053);cAMP ELISA试剂盒(南京建成生物工程研究所,批号:20210611);Fluo-3/AM钙离子荧光探针(上海翊圣生物科技有限公司,批号:F2023461);Lactisole(上海源叶生物科技有限公司,批号:L06J12T136516)。
TCSSP5Ⅱ激光共聚焦荧光显微镜(德国Leica公司);Eclipse TS100-F倒置显微镜(日本Nikon公司);ACCUCHEKN Active BloodGlucose Meter(罗氏活力型血糖仪);HI1220烘片机(德国Leica公司);EG1150H石蜡包埋热台(德国Leica公司);RM2255轮转式病理切片(德国Leica公司);MODEL 3111二氧化碳培养箱(Thermo e-lectron公司)。
取玉竹,粉碎,加水提取3次,每次2 h;合并提取液,醇沉,过滤得上清液;上清液浓缩,乙酸乙酯萃取,收集乙酸乙酯,蒸干得干膏;干膏用水溶解,AB-8大孔树脂纯化,先用水、10%乙醇除去杂质,再用40%乙醇洗脱。收集洗脱液,蒸干,即得玉竹皂苷[11]。经测定,玉竹皂苷纯度为63.5%,得率为0.1%,且不含有玉竹中含有的葡萄糖、蔗糖、果糖、鼠李糖、木糖等小分子糖类成分。
动物购回后适应性喂养1周后,禁食过夜,再以120 mg/kg的1% STZ一次性腹腔注射制备糖尿病模型,正常组(normal control group,NC组)注射等剂量空白缓冲液。3 d后,尾静脉法取血,测定随机血糖,若≥16.7 mmol/L,表明造模成功[12]。将糖尿病小鼠随机分成6组,分别为模型组(diabetes model group,DM组)、阳性组(positive control group,PC组,二甲双胍250 mg/kg)、玉竹皂苷低剂量组(low dose group,LD组,18.2 mg/kg)、玉竹皂苷高剂量组(high dose group,HD组,72.8 mg/kg)、玉竹皂苷低剂量+100 mg/kg Lactisole组(low dose group+Lactisole,LD+Lac组)、玉竹皂苷高剂量+100 mg/kg Lactisole组(high dose group+Lactisole,HD+Lac组),每组6只。给药组给予相应药物,正常组、模型组给予相应体积的蒸馏水,持续8周。
分别于给药前及给药后2、4、6、8周进行体质量、摄食量、摄水量、尿量的测定。将小鼠放入代谢笼,每笼放入一定量的饲料(W1)与水(V1),收集24 h尿液,24 h后称量剩余饲料(W2)与水(V2),将收集的尿液离心,分离尿液,测定尿量[13]。摄食量公式:摄食量=W1-W2;摄水量公式:摄水量=V1-V2。
给药7周后,禁食过夜,尾静脉取血测空腹血糖[14-15]。禁食16 h后,以2.0 g/kg葡萄糖溶液灌胃,尾静脉取血测灌胃后0、0.5、1.0、1.5、2.0 h的血糖值,并计算其血糖曲线下面积(area under the curve,AUC)[16]。
给药8周结束后,禁食过夜;麻醉,解剖,收集小鼠胰腺、肝脏等组织;放入4% 多聚甲醛中固定24 h,乙醇脱水,二甲苯透明,浸蜡包埋切片,用中性树脂封片,置于光学显微镜下观察。
给药8周结束后,小鼠禁食过夜,麻醉后摘眼球取血,血液静置2 h后,4℃,3500 r/min离心10 min,分离血清;按相应试剂盒说明书操作,测定TC、TG、LDL-C、HDLC的含量。
取“2.3.4”项下的血清,ELISA测定其中GLP-1、胰岛素的含量。
培养HuTu-80细胞,取对数生长期细胞,以每孔4×105个接种到6孔板中,设置正常组,葡萄糖组(300μg/mL),100、200、300、400、500μg/mL玉竹皂苷组;细胞置于CO2培养箱中培养48 h后,将培养液吸去,磷酸缓冲盐溶液(phosphate buffered saline,PBS)清洗细胞2次;再加入无糖培养液继续培养3 h;吸去培养液,PBS清洗2次,分别加入相应药物,孵育1 h。吸取细胞培养液于EP管内,4℃离心10 min,取上清液,ELISA法检测各组GLP-1含量;然后在300μg/mL玉竹皂苷中分别加入5 mmol/L Lactisole、10 mmol/L Lactisole,考察玉竹皂苷与Lactisole合用对GLP-1分泌的影响。
取对数生长期细胞,以每孔4×105个接种到6孔板中,设置正常组、葡萄糖组(300μg/mL)、玉竹皂苷组(300μg/mL)。细胞置于CO2培养箱中培养48 h,将培养液吸去,PBS清洗2次;加入无糖培养液,放入培养箱中继续培养3 h。吸去培养液,PBS清洗细胞2次,再分别加入相应药物,正常组加入同体积的细胞培养液,放入培养箱孵育15 min。吸去培养液,PBS清洗2次,加入RIPA裂解液,消化2 min后吹打均匀,4℃离心10 min,收集上清液,ELISA法检测胞内cAMP的含量。
取对数生长期细胞,以每孔1×105个加入到激光共聚焦培养皿底孔中,静置,贴壁,再孵育24 h;设置正常组、300μg/mL葡萄糖组、300μg/mL玉竹皂苷组。将培养液吸去,PBS清洗2次,加入无糖培养液,放入培养箱中继续培养3 h。吸去培养液,PBS清洗2次后,避光加入2μmol/L Fluo-3/AM,每孔150μL;37℃孵育10 min,PBS清洗2次后,给药组加入相应药物,正常组加入同体积的细胞培养液,培养箱中孵育5 min,吸去培养液,PBS清洗2次;再加入PBS缓冲液,放入培养箱继续孵育15 min。用激光共聚焦显微镜观察并拍照,采用Image Pro Plus软件计算Ca2+荧光强度[17]。
实验数据用表示,采用SPSS19.0统计软件进行数据处理,组间均数比较时用单因素方差分析,组间均数两两比较采用最小显著差数(LSD)检验,方差不齐时用Tamhane检验,以P<0.05表示差异具有统计学意义。
玉竹皂苷对小鼠体质量、摄食量、摄水量、尿量的影响,结果见图1。图1A显示,与模型组比较,玉竹皂苷低、高剂量组小鼠在治疗6周后,体质量明显增加(P<0.05)。图1B显示,与正常组比较,2周后模型组小鼠摄食量显著升高(P<0.05);与模型组比较,玉竹皂苷低、高剂量组小鼠在治疗6、8周后,摄食量明显降低(P<0.05)。图1C显示,与模型组比较,玉竹皂苷低、高剂量组小鼠在治疗8周后,摄水量明显下降(P<0.05)。图1D显示,与模型组比较,玉竹皂苷低、高剂量组小鼠在治疗6周后,尿量显著减少(P<0.05)。然而,玉竹皂苷对糖尿病小鼠摄水量、尿量的作用,均可被甜味受体抑制剂Lactisole抑制(P<0.05)。
玉竹皂苷对小鼠空腹血糖的影响,结果见插页Ⅰ图2A。结果显示,与正常组比较,模型组小鼠空腹血糖显著升高(P<0.05),玉竹皂苷治疗6周后,空腹血糖明显下降(P<0.05);玉竹皂苷对糖尿病小鼠OGTT的影响,结果见插页Ⅰ图2B。结果显示,与正常组比较,模型组小鼠血糖曲线下面积显著增大(P<0.05),给予玉竹皂苷后,血糖曲线下面积明显减小(P<0.05)。插页Ⅰ图2A-图2B结果同时表明,Lactisole可逆转玉竹皂苷对空腹血糖、口服葡萄糖耐量的作用(P<0.05)。
玉竹皂苷对胰腺、肝组织形态的影响,结果见插页Ⅱ图3。结果显示,正常组小鼠的胰岛组织结构完整,边界清晰,细胞分布均匀,模型组小鼠胰岛组织萎缩变形,边界不规则,而玉竹皂苷可明显修复胰岛组织的损伤。同时,正常小鼠肝细胞分布密集,细胞形态良好,肝细胞及肝索在肝静脉周围呈放射状排列;模型组小鼠肝组织肝索消失,肝细胞内有大空泡形成,细胞核变大,并出现炎细胞浸润;给予玉竹皂苷后,肝细胞形态有所改善,炎细胞浸润减少。而Lactisole可逆转玉竹皂苷对胰腺、肝脏组织的作用,使其损伤加重。
玉竹皂苷对小鼠血脂的影响,结果见表1。结果显示,与正常组比较,模型组小鼠TC、TG、LDL-C水平显著升高(P<0.05),HDL-C水平显著下降(P<0.05);与模型组比较,玉竹皂苷低、高剂量组小鼠血清TC、TG水平明显下降(P<0.05),HDL-C水平显著升高(P<0.05);同时给予玉竹皂苷与甜味受体抑制剂Lactisole,Lactisole可抑制玉竹皂苷对TC、TG、HDL-C的作用(P<0.05)。
玉竹皂苷对GLP-1的影响,结果见图4A。结果显示,与正常组比较,模型组GLP-1含量明显下降(P<0.05);与模型组比较,玉竹皂苷组GLP-1含量显著升高(P<0.05)。玉竹皂苷对胰岛素的影响,结果见图4B。结果显示,与正常组比较,模型组小鼠血清胰岛素含量显著下降(P<0.05)。与模型组比较,玉竹皂苷组小鼠血清胰岛素含量显著升高(P<0.05)。图4也表明,Lactisole可逆转玉竹皂苷对GLP-1、胰岛素的作用(P<0.05)。
玉竹皂苷对HuTu-80细胞GLP-1分泌的影响,结果见插页Ⅰ图5A-图5B。插页Ⅰ图5A显示,与正常组相比,200~500μg/mL的玉竹皂苷均能显著增加胞外GLP-1的浓度(P<0.05);插页Ⅰ图5B显示,Lactisole能明显下调玉竹皂苷对GLP-1分泌的促进作用(P<0.05)。进一步研究玉竹皂苷对甜味受体信号通路信号分子cAMP、Ca2+的影响,结果见插页Ⅰ图5C-图5D。插页Ⅰ图5C显示,玉竹皂苷可显著增加细胞内甜味受体信号通路信号分子cAMP含量(P<0.05);插页Ⅰ图5D表明,与正常组(0.06×10-1±0.02×10-1)相比,玉竹皂苷组(0.31×10-1±0.07×10-1)及葡萄糖组(0.16×10-1±0.03×10-1)中细胞内Ca2+荧光强度明显增强(P<0.05)。
本研究采用一次性腹腔注射STZ的方法建立糖尿病模型,糖尿病小鼠经玉竹皂苷治疗后,其摄食量、摄水量、尿量、TC、TG、HDL-C、空腹血糖、口服葡萄糖耐量以及胰腺、肝组织形态学均发生显著变化,表明玉竹皂苷有明显的降血糖效应,对糖尿病也有一定的治疗作用。同时,与模型组比较,玉竹皂苷组小鼠的GLP-1、胰岛素的浓度显著升高,且甜味受体抑制剂Lactisole能明显阻断玉竹皂苷对TC、TG、HDL-C、GLP-1、胰岛素等指标的作用。可见,玉竹皂苷是通过甜味受体发挥降血糖作用。换而言之,激动甜味受体可发挥降血糖作用。进一步采用HuTu-80细胞验证玉竹皂苷对甜味受体的激动作用。HuTu-80细胞为十二指肠腺癌细胞,其中有甜味受体的表达,与甜味受体激动剂结合后,cAMP分泌增加,Ca2+内流,进而GLP-1分泌增加。细胞实验结果再次证实,玉竹皂苷可激动甜味受体,启动甜味受体信号通路。
目前,糖尿病主要治疗方法是降血糖,GLP-1受体激动剂是一类降血糖药物,如艾塞那肽[18-19]。GLP-1是一种内源性肠促胰岛素激素,其通过促进胰岛素释放、抑制胰高血糖素释放、抑制食欲和胃排空来降低血糖[20-21]。本研究发现,玉竹皂苷可与甜味受体结合,启动甜味受体信号通路,进而促进GLP-1的分泌,最终导致胰岛素分泌增加。可见,GLP-1受体激动剂与甜味受体激动剂在促进胰岛素分泌、降血糖等方面其作用是一致的。本项目前期研究也表明,玉竹皂苷与甜味受体有较好的结合性[11]。
本文针对目前中药甘味的内涵,结合甜味受体,提出“凡可启动、增强甜味信号的物质,均为甘味”。这种新界定的“甘味”不仅包括滋味,也包括效味。其一,这种新界定的中药“甘味”,有“滋味”。基于滋味确定的甘味中药,其本质上是其中含有葡萄糖、蔗糖等甜味受体激动剂。其二,这种新界定的中药“甘味”,也有“效味”。启动、增强甜味信号的物质,均可直接或间接产生GLP-1。传统中医药理论表明,甘味药能补、能和、能缓,即甘味药具有补益、和中、调和药性等作用[22-23]。其中,补益作用又包括补气、补阳、补阴、补血。气属阳,血属阴,因此,甘味药的补益作用可进一步归纳为补阴、补阳。启动甜味受体信号通路,GLP-1分泌增加,进而促进胰岛素分泌,在降低血糖的同时,通过调节血液渗透压进一步影响水液代谢,而阴虚、阳虚证的水液代谢均可能出现紊乱。因此,这种新界定的中药“甘味”,在一定程度上可阐释甘味药的“补益”作用。需要指出的是,基于新界定的中药“甘味”阐释甘味药和中、调和药性等效应,还需进一步分析。
玉竹常用于治疗阴虚证,现代用于治疗糖尿病[24]。糖尿病有阴虚表现,阴虚也可能导致糖尿病[25-26]。“甘补阴”,因此玉竹治疗糖尿病可从“甘味”的角度探讨。基于新界定的中药“甘味”,自然可从甜味受体的角度探讨玉竹降血糖、治疗糖尿病的机制。实验结果表明,从甜味受体的角度能较好地阐释玉竹活性成分——玉竹皂苷的降血糖机制。因此,这种新界定的中药“甘味”,在实践上也有可行性。总之,本文基于甜味受体整合滋味、效味,提出界定中药“甘味”新方法,为中药的五味研究提供了新视角,助推了五味理论的传承与发展。同时,研究结果也为玉竹的应用及评价提供实验依据。
  • 国家自然科学基金项目(81960724)
  • 江西省自然科学基金面上项目(20232BAB206171)
  • 江西省大学生创新创业训练计划项目(S202310412101)
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2025年第43卷第7期
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doi: 10.13193/j.issn.1673-7717.2025.07.001
  • 首发时间:2026-10-30
  • 出版时间:2025-07-10
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基金
国家自然科学基金项目(81960724)
江西省自然科学基金面上项目(20232BAB206171)
江西省大学生创新创业训练计划项目(S202310412101)
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    江西中医药大学,江西 南昌 330004

通讯作者:

吴璐(1980-),女,湖南长沙人,副教授,硕士,研究方向:传统中医药理论及中药制剂。E-mail:。
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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