Identification of Polygonatum from Wuyishan and Its Surrounding Areas by ISSR Molecular Markers and Their HPLC Fingerprints

采用ISSR分子标记鉴定方法及HPLC色谱方法对武夷山及周边地区5份黄精植物样本进行DNA分子标记鉴定及指纹图谱分析,并通过聚类分析软件分别探讨其遗传相关聚类图谱。ISSR分子标记法能较好地对5份黄精植物样本进行区分,HPLC指纹图谱分析同样表明5份黄精样本化学成分存在一定的差别。DNA分子标记及HPLC指纹图谱分析的遗传相关聚类分析结果相似,表明武夷山及周边地区的野生黄精种质资源存在明显的差别。The purpose of this project was to identify and analyze the genetic relationships of 5 kinds of Polygonatum in Wuyishan and its surrounding areas. Using ISSR marker and HPLC molecular identification method for chromatographic methods analysis of DNA markers for identification and fingerprinting of 5 Polygonatum samples, the genetic correlation map of Polygonatum was studied by cluster analysis software. ISSR molecular marker method could be used to distinguish Polygonatum of Wuyishan and its surrounding areas. HPLC fingerprint analysis also showed that there were some differences in the composition of Wuyishan and its surrounding distribution. The results of genetic correlation cluster analysis were similar by ISSR and HPLC. There were obvious differences in the germplasm resources of Wuyishan and its surrounding areas from the DNA molecular markers and HPLC fingerprinting analysis results.福建省科技厅自然科学基金重点项目（2013Y0018）;福建省科技厅自然科学基金青年项目（2015J01065

没食子儿茶素促进脂肪细胞吸收葡萄糖研究

目的探讨没食子儿茶素对脂肪细胞在高浓度葡萄糖条件下吸收葡萄糖的作用。方法应用3T3-L1前脂肪细胞分化模型,测定在脂肪细胞中没食子儿茶素对葡萄糖吸收的作用和对3T3-L1前脂肪细胞分化的影响。结果没食子儿茶素可促进脂肪细胞中高浓度葡萄糖(30 mmol/L)条件下由胰岛素刺激的葡萄糖吸收,加快3T3-L1前脂肪细胞的分化。结论没食子儿茶素可增加脂肪细胞葡萄糖吸收并促进3T3-L1前脂肪细胞分化,提示没食子儿茶素可有效改变血糖浓度和改善胰岛素抵抗。国家自然科学基金面上项目(81773601);;福建省卫生系统中青年骨干人才培养项目2015年资助计划基础项目;;厦门市卫计委资助项目(2015-ZQN-JC-45);;福建省自然科学基金计划资助项目(2015J01065);;福建省教育厅2015年第二批高等学校优秀学科(专业)带头人赴海外访学研修资助项目[闽教人(2015)103号];;福建省厦门市科技局科技计划高校创新项目(3502Z20143026);福建省厦门市科技局科技计划项目(3502Z20144031);;厦门医学院科研项目(Z2013-12,Z2013-25

Effects of the Xitongxiao prescription on apoptosis and proliferation of chondrocytes in knee

目的：探讨膝痛消方对促进软骨细胞增殖和抑制IL-1β诱导的软骨细胞凋亡的作用。方法：体外培养膝关节软骨细胞，用阿利新蓝染色方法检测软骨细胞蛋白多糖，用Ⅱ型胶原免疫荧光染色检测软骨细胞Ⅱ型胶原的表达，以鉴定软骨细胞；用MTS检测软骨细胞的增殖；用IL-1β诱导软骨细胞凋亡，AnnexinV/PI双染检测细胞凋亡。结果：膝关节剥离得到的细胞形态为多角形或梭形，阿利新蓝与Ⅱ型胶原免疫荧光染色均呈阳性，证明分离培养的细胞为软骨细胞。MTS实验结果发现膝痛消方提取物处理软骨细胞后检测到的OD值随浓度增加而增加且显著高于对照组。此外，IL-1β处理诱导软骨细胞凋亡，而膝痛消方提取物能够有效地抑制IL-1β引起的细胞凋亡。结论：膝痛消方能促进软骨细胞的增殖，抑制IL-1β诱导的软骨细胞的凋亡。以上发现为膝痛消方临床上治疗膝关节骨性关节炎提供药理作用依据。Objective:To investigate the effects of the Xitongxiao prescription on the cell proliferation and the inhibition of IL-1β-induced apoptosis of chondrocytes.Methods:To identify chondrocytes,the articular chondrocytes were cultured in vitro;staining with alcian blue was used to detect the protein polysaccharide,and staining with collagen immunofluorescence was used to examine the expression of type II collagen in chondrocytes.The proliferation of chondrocytes was determined by MTS.Apoptosis of chondrocytes was induced by IL-1β,and the effect of apoptosis was tested by Annexin V/PI double staining.Results:The morphology of the cells obtained from the knee joint was polygonal or fusiform.The tested results of Alcian blue and collagen immunofluorescence staining were positive.These together certified that the isolated cells were knee chondrocytes.MTS assay results showed that the detected OD values were dosedependently increased and they were significantly higher than those of the control group.In addition,the apoptosis of chondrocytes was induced by IL-1βtreatment.While the extract of the Xitongxiao prescription could effectively inhibit the IL-1β-induced apoptosis of chondrocytes.Conclusion:The Xitongxiao prescription could promote the proliferation of chondrocytes and inhibit the apoptosis of chondrocytes induced by IL-1β.The above findings may provide one of pharmacological mechanisms of the Xitongxiao for clinical treatment of knee osteoarthritis.厦门市科技计划项目(3502z20144031);福建省自然科学基金计划资助项目(2015J01065)

Screening research on effective parts of Alisma orientalis( Samuel. ) Juzep. associated with blood glucose decrease in type 2 diabetic mice

目的初步探索泽泻[Alisma orientalis(Samuel.)Juzep.]降低2型糖尿病小鼠血糖的有效部位。方法采用超声提取法获得泽泻乙醇提取物,再分别用石油醚、乙酸乙酯、水饱和正丁醇和水相进行萃取分离,得到的泽泻乙醇提取物及其各溶剂萃取物应用于腹腔注射烟碱、链脲霉素诱导的2型糖尿病小鼠模型。造模成功后,将动物分组,其中空白对照组(未注射链脲霉素和烟碱的小鼠)和2型糖尿病模型组小鼠连续灌胃0.9%Na Cl溶液21 d(灌胃剂量为10μl/g),测试药物组分别连续灌胃泽泻乙醇提取物、泽泻石油醚萃取物、泽泻乙酸乙酯萃取物、泽泻正丁醇萃取物、泽泻水萃取物21 d(各测试药物灌胃高、中、低剂量分别为100μg/g、50μg/g、25μg/g),阳性对照组连续灌胃罗格列酮溶液21 d(灌胃剂量为10μg/g),每组8只小鼠,筛选具有降低2型糖尿病小鼠血糖的泽泻有效部位。用具有降低2型糖尿病小鼠血糖的有效部位处理3T3-L1前脂肪细胞,观察其对前脂肪细胞分化过程的影响。结果与2型糖尿病模型组比较,连续灌胃泽泻乙酸乙酯萃取物21 d(剂量为100μg/g)降低了2型糖尿病小鼠的血糖,改善了口服葡萄糖耐受试验(oral glucose tolerance test,OGTT)过程中的糖耐量(P<0.01)。1 mg/L和10 mg/L的泽泻乙酸乙酯萃取层均能促进前脂肪细胞的分化过程。结论泽泻乙酸乙酯萃取物降低了2型糖尿病小鼠血糖,促进了3T3-L1前脂肪细胞分化,但其具体作用机制尚需深入研究。Objective To explore the effective parts of Alisma orientalis( Samuel.) Juzep. associated with blood glucose decrease in type 2 diabetic mice.Methods Ethanol extracts from A. orientalis were obtained by ultrasonic extraction. Then,the ethanol extract was extracted and separated with petroleum ether,ethyl acetate,water saturated n-butanol and water respectively. After decompressing concentration,these solvent extractions were applied to type 2 diabetic mice model induced by intraperitoneal injection of streptozotocin and nicotine. After successful modeling,the mice were divided into the blank control group( without injection of streptozotocin and nicotine),model group of type 2 diabetes mellitus,positive control group and test drug groups,8 in each group. The blank control group and model group were treated with 0. 9% Na Cl at the dose of 10μl / g,the test drug groups were treated with corresponding extracts from different solvent extractions described above at the high-,mid-,low-dose of 100,50,25 μg / g respectively. The positive control group was treated with rosiglitazone at the dose of10μg / g. The treatment course was 21 days for intragastric administration. The effective parts of Alisma orientalis( Samuel.) Juzep. associated with blood glucose decrease in type 2 diabetic mice were screened and then the effect of the screened effective part on the differentiation of 3T3-L1 pre-adipocyte was observed.Results Compared with the model group,the blood glucose was decreased after the intragastric administration with ethyl acetate extract of A. orientalis at dose of100μg / g for 21 days and the glucose tolerance during the oral glucose tolerance test( OGTT) was improved( P < 0. 01). The differentiation of pre-adipocyte was promoted when the ethyl acetate extract of A. orientalis at the concentration of 1 mg / L and 10 mg / L. Conclusion The ethyl acetate extract of A. orientalis reduced the blood glucose in type 2 diabetic mice and promoted the differentiation of 3T3-L1 pre-adipocytes,but the specific mechanism still need to be further studied.福建省科技厅自然科学基金青年创新项目;; 厦门市卫计委资助项目(2014D008),厦门市卫计委资助项目(2015-ZQN-JC-45);; 福建省卫生系统中青年骨干人才培养项目基础项目;; 福建省科技厅自然科学基金(2015J01065);; 福建省教育厅中青年教师教育科研项目杰青项目(JA14421),福建省教育厅中青年教师教育科研项目(JA13425);; 厦门市科技局科技计划高校创新项目(3502Z20143026),厦门市科技局科技计划项目(3502Z20144031);; 厦门医学高等专科学校科研基金(Z2013-12,Z2013-25

太子参环肽类化合物Pseudostellarin E对3T3-L1前脂肪细胞分化和葡萄糖吸收的作用

目的探讨太子参环肽类化合物Pseudostellarin E是否具有治疗2型糖尿病的潜力。方法在3T3-L1前脂肪细胞中应用了葡萄糖吸收试验和分化模型。结果 Pseudostellarin E不仅加速3T3-L1前脂肪细胞分化进程,而且在高浓度(30m M)葡萄糖条件下增加了分化的脂肪细胞对胰岛素刺激的葡萄糖的吸收。结论上述结果表明,Pseudostellarin E可能具有治疗糖尿病的潜能。国家自然科学基金面上项目(81773601);;福建省教育厅2015年第二批高等学校优秀学科(专业)带头人赴海外访学研修资助项目(闽教人[2015]103号);;福建省自然科学基金计划资助项目(2015J01065);;福建省卫生系统中青年骨干人才培养项目2015年资助计划基础项目(2015-ZQN-JC-45);;福建省厦门市科技局科技计划高校创新项目(3502Z20143026);;厦门医学院科研项目(Z2013-12,Z2013-25

Effect of Ethanol Extract from Camellia Oleifera Abel. on Blood Glucose in Type 2 Diabetic Mice

目的筛选油茶（Camellia oleifera Abel.）降低2型糖尿病小鼠血糖的有效部位。方法采用腹腔注射烟碱、链脲霉素诱导的2型糖尿病小鼠模型,应用超声提取法分别获得油茶叶、果、籽的乙醇提取物,筛选降低血糖的有效部位。结果阳性药罗格列酮每日灌胃剂量10mg/kg·BW,连续灌胃21d后,降低了2型糖尿病小鼠血糖。分别灌胃油茶叶、油茶果、油茶籽乙醇提取物,按每日剂量100mg/kg·BW连续灌胃21d后,均在一定程度上降低2型糖尿病小鼠血糖,油茶籽乙醇提取物对口服葡萄糖耐受试验（Oral glucose tolerance test,OGTT）作用最明显。与对照组相比,1μM和2μM的阳性药罗格列酮均能明显促进3T3-L1前脂肪细胞分化;油茶籽乙醇提取物在0.5μg·m L-1和1μg·m L-1浓度下也体现出促进分化的活性。结论油茶籽乙醇提取物降低了2型糖尿病小鼠的血糖,促进了3T3-L1前脂肪细胞的分化,但其具体作用机制尚需深入研究。OBJECTIVE To investigate effective parts possessing property of lowering blood glucose in type 2diabetic mice from Camellia oleifera Abel. METHODS Leaf,fruit and seed ethanol extract of C. oleifera were obtained through ultrasonic extraction,and were subsequently subject to type 2 diabetic mice model induced by intraperitoneal injection of streptozocin and nicotine. RESULTS In accordance with the positive drug rosiglitazone by intragastric administration once daily dose of 10 mg / kg·BW,continuous intragastric administration for 21 days,blood glucose in type 2 diabetic mice was reduced. In accordance with a daily dose of 100 mg / kg·BW leaf,fruit and seed ethanol extract of C. oleifera,continuous intragastric administration for 21 days respectively,in a certain extent,blood glucose in type 2 diabetic mice was reduced. Moreover,effect of seed ethanol extract on blood glucose and oral glucose tolerance test（ OGTT） is the most obvious. Compared with control group（ treated with insulin）,1μM and 2μM of positive drug rosiglitazone can significantly promote differentiation of 3T3-L1 pre-adipocytes,C. oleifera seed ethanol extract as the most effective part for lowing blood glucose at 0. 5μg·m L~（-1） and 1μg·m L~（-1）concentration also promoted 3T3-L1 pre-adipocyte differentiation. CONCLUSION It was suggested that C. oleifera seed ethanol extract lowered blood glucose in type 2 diabetic mice and enhanced differentiation of 3T3-L1 pre-adipocytes,but the specific mechanism needs further study.福建省教育厅中青年教师教育科研项目杰青项目JA14421;福建省教育厅中青年教师教育科研项目JA13425; 厦门市科技局科技计划高校创新项目3502Z20143026;厦门市科技局科技计划项目3502Z20144031; 福建省自然科学基金计划资助项目2015J01065; 福建省科技厅自然科学基金青年创新项目2014D008; 福建省卫生系统中青年骨干人才培养项目基础项目2015-ZQN-JC-45; 厦门市卫生和计划生育委员会资助研究经费; 厦门医学高等专科学校科研基金Z2013-12;Z2013-2

23-乙酰泽泻醇B对2型糖尿病小鼠血糖的影响

目的 探讨23-乙酰泽泻醇B是否有治疗2型糖尿病的潜能。方法 通过腹腔注射链脲佐菌素和烟酰胺建立2型糖尿病小鼠模型。灌胃罗格列酮或23-乙酰泽泻醇B 3周后,测定2型糖尿病小鼠血糖值,次日进行口服葡萄糖耐受试验（OGTT）。采用葡萄糖荧光示踪剂,测定23-乙酰泽泻醇B对葡萄糖吸收的影响。采用3T3-L1前脂肪细胞分化模型,测定其对分化的影响。结果 分别每天灌胃阳性药罗格列酮10 mg·kg-1、23-乙酰泽泻醇B(5、10、20 mg·kg-1),连续灌胃给药3周后,降低了2型糖尿病小鼠血糖值,一定程度改善OGTT过程中胰岛素抵抗。在30 mmol·L-1高糖条件下,23-乙酰泽泻醇B促进了脂肪细胞对胰岛素刺激的葡萄糖吸收; 23-乙酰泽泻醇B(1、10μmol·L-1)促进3T3-L1前脂肪细胞的分化过程。结论 23-乙酰泽泻醇B降低2型糖尿病小鼠血糖,促进前脂肪细胞分化,促进脂肪细胞吸收葡萄糖,但作用机制仍需进一步探索。国家自然科学基金面上项目(No 81773601);;福建省卫生系统中青年骨干人才培养项目(No 2015-ZQN-JC-45);;福建省教育厅中青年教师教育科研项目(No JT180664

槐花提取物降血糖活性研究

目的研究槐花Sophora japonica L.(Leguminosae)提取物对2型糖尿病小鼠降血糖活性。方法通过超声提取法获得槐花乙醇提取物。使用腹腔注射烟碱和链脲霉素诱导的2型糖尿病小鼠模型和3T3-L1前脂肪细胞分化模型,检测槐花乙醇提取物对小鼠血糖的影响和对3T3-L1前脂肪细胞分化的影响。结果槐花乙醇提取物灌胃剂量每日100μg/g,连续灌胃给药3周,降低了2型糖尿病小鼠的血糖,改善口服葡萄糖耐受试验OGTT过程中的糖耐量。0.4 mg/L和0.8 mg/L的槐花乙醇提取物均能促进3T3-L1前脂肪细胞分化。结论槐花乙醇提取物降低2型糖尿病小鼠血糖,促进3T3-L1前脂肪细胞分化,具体作用机制有待进一步研究。福建省教育厅中青年教师教育科研项目(JA13425);福建省教育厅中青年教师教育科研项目杰青项目(JA14421);福建省卫生系统中青年骨干人才培养项目基础项目;厦门市卫计委资助项目(2015-ZQN-JC-45);厦门市卫计委资助项目(2014D008);福建省科技厅自然科学基金青年创新项目;福建省科技厅自然科学基金项目(2015J01065);厦门市科技局科技计划高校创新项目(3502Z20143026);厦门市科技局科技计划项目(3502Z20144031);厦门医学院科研基金(Z2013-12、Z2013-25

23-乙酰泽泻醇B对2型糖尿病小鼠血糖的影响

目的探讨23-乙酰泽泻醇B是否有治疗2型糖尿病的潜能。方法通过腹腔注射链脲佐菌素和烟酰胺建立2型糖尿病小鼠模型。灌胃罗格列酮或23-乙酰泽泻醇B 3周后,测定2型糖尿病小鼠血糖值,次日进行...国家自然科学基金面上项目(No 81773601);; 福建省卫生系统中青年骨干人才培养项目(No 2015-ZQN-JC-45);; 福建省教育厅中青年教师教育科研项目(No JT180664