Dangnyohwan improves glucose utilization and reduces insulin resistance by increasing the adipocyte-specific GLUT4 expression in Otsuka Long-Evans Tokushima Fatty rats

AIM OF THE STUDY: Dangnyohwan (DNH) has been used for treatment of diabetes mellitus. However, the exact cellular and molecular mechanisms underlying the beneficial effects of DNH are not well understood. Therefore, we investigated how DNH improves hyperglycemia and insulin resistance in obese-type diabetes model. METHODS AND MATERIALS: We examined the effect of DNH on the expression of glucose transporter 4 (GLUT4), GLUT4 translocation, and glucose transport activity in muscle and adipose tissues from Otsuka Long-Evans Tokushima Fatty (OLETF) and Long-Evans Tokushima Otsuka (LETO) rats. RESULTS: DNH ameliorated hyperglycemia and impaired glucose tolerance (IGT) observed in 26- and 42-week-old male OLETF rats. The basal and insulin-stimulated [14C]2-Deoxyglucose (2DG) uptake was significantly increased in adipocytes from DNH-treated OLETF rats, as compared with untreated OLETF rats. The expression level of GLUT4 was markedly decreased (by 90-95%) in the adipose tissue of OLETF rats, whereas DNH treatment drastically increased the expression of GLUT4 within 8 weeks. DNH improved GLUT4 recruitment stimulated by insulin in both the 26- and 42-week-old OLETF rat adipocytes. CONCLUSION: These results suggest that DNH could exert the beneficial effects on hyperglycemia and insulin resistance by increasing the expression and insulin-stimulated translocation of GLUT4 in OLETF rat adipocytes.     

Coptidis Rhizoma Water Extract Stimulates 5'-AMP-Activated Protein Kinase in Rat Skeletal Muscle

AIM:Coptidis Rhizoma(CR),the dried rhizomes of Asian herbs(including Coptis chinensis Franch),has been used to treat diabetes mellitus for thousands of years.We explored the possibility that CR acts directly on skeletal muscle,the major organ responsible for glucose homeostasis,and activates 5'-AMP-activated protein kinase(AMPK),a signaling intermediary leading to metabolic enhancement of skeletal muscle.METHODS:Isolated rat epitrochlearis and soleus muscles were incubated in a buffer containing a CR water ...     

泻心汤配伍理论实验研究

目的: 探讨泻心汤配伍合理性及其可能机制。 方法: 运用正交设计探讨泻心汤各组分对小鼠的急性毒性作用及相互关系。 结果: 体内研究发现黄连、大黄对小鼠ig均有一定的急性毒性作用,测得其小鼠ig大黄的半数致死量(LD₅₀)按生药计为48.92 g/kg;小鼠ig黄连的LD₅₀按生药计为2 047.9 mg/kg;黄芩对小鼠无明显急性毒性作用,未测出其小鼠口服LD₅₀。利用正交设计,以死亡率为指标观察泻心汤各组分对小鼠的急性毒性作用,结果显示:黄连是泻心汤中的主要有毒物质;大黄有降低黄连急性毒性作用的趋势,但作用明显弱于黄芩;黄芩可显著降低黄连的急性毒性,是泻心汤中的主要解毒物质,这可能是泻心汤在临床应用中表现出较小毒性的重要原因。 结论: 泻心汤中黄连具有一定的急性毒性,黄芩则是其解毒物质,该方的配伍从某种程度上达到了解毒的目的,具有良好的科学性,其解毒的机制可能与黄连中的生物碱能与黄芩苷或大黄蒽醌生成沉淀有关。     

四种中药对感染牙本质小管粪肠球菌的抑制作用

目的比较黄连等4种中药水煎剂对离体人牙根管内粪肠球菌的体外抑菌效果。方法选取84颗新鲜拔除的人单根管牙,截取6mm高的柱状牙根。随机分为4个实验组(黄连水煎剂组、五味子水煎剂组、大黄水煎剂组、连翘水煎剂组)和3个对照组[甲醛甲酚(FC)组、阴性对照组和阳性对照组],每组12颗牙。常规根管预备后灭菌制备感染粪肠球菌根管模型,阴性对照组不感染细菌,放置0.9%氯化钠注射液,余每组根管均感染细菌,分别放置上述不同药剂,3天、7天后分别培养并测量细菌的透光度。结果黄连水煎剂组3天与甲酫甲酚组3天组间无差别(P>0.05);连翘水煎剂组3天与阳性对照组3天组间无差别(P>0.05);五味子水煎剂组3天与阴性对照组7天组间无差别(P>0.05);大黄水煎剂组3天、黄连水煎剂组7天与甲酫甲酚组7天组间无差别(P>0.05);连翘水煎剂组7天与阳性对照组7天组间无差别(P>0.05);其余各组间差别均有统计学意义(P<0.05)。结论四种中药中,黄连、五味子、大黄均有不同程度抑制粪肠球菌的作用,黄连作用最强。黄连抑菌作用与FC相同。连翘无抑制粪肠球菌的作用。     

三清降糖方对代谢综合征大鼠糖脂代谢、血清瘦素及游离脂肪酸的影响

目的观察三清降糖方对代谢综合征大鼠胰岛素抵抗、血脂、血清瘦素及游离脂肪酸的影响。方法采用高脂高糖高盐饮食喂养方法制备大鼠代谢综合征模型,并随机分为正常组、模型组、罗格列酮组和三清降糖方低、中、高组。灌胃给药4周,采用生化法测血糖血脂,放免法测胰岛素及血清瘦素水平,比色法测游离脂肪酸水平,计算胰岛素抵抗指数。结果三清降糖方各组及罗格列酮组血脂、血糖、血清胰岛素、瘦素及游离脂肪酸等指标与模型组相比均显著降低(P均<0.05),三清降糖方各组与罗格列酮组比较无显著性差异。结论三清降糖方可以改善代谢综合征大鼠胰岛素抵抗,调节血脂,可能与降低血清瘦素及游离脂肪酸水平有关。     

A refined-JinQi-JiangTang tablet ameliorates prediabetes by reducing insulin resistance and improving beta cell function in mice

Ethnopharmacological relevance

Refined-JQ (JQ-R) is a mixture of refined extracts from three major herbal components of JinQi-JiangTang tablet: Coptis chinensis (Ranunculaceae), Astragalus membranaceus (Leguminosae), and Lonicera japonica (Caprifoliaceae). Our previous studies have indicated that JQ-R could decrease fasting blood glucose levels in diabetic mice and insulin resistance mice. Investigating the hypoglycemic effect of JQ-R on prediabetes has practical application value for preventing or delaying insulin resistance, impaired glucose tolerance and possibly the development of clinical diabetes.

Materials and methods

The anti-diabetic potential of JQ-R was investigated using a high fat-diet (HFD)-induced obesity mouse model. C57BL/6J mice (HFD-C57 mice) were fed with high-fat diet for 4 months. HFD-C57 mice were treated with either JQ-R (administered intragastrically once daily for 4 weeks) or metformin (as positive control), and the effects of JQ-R on body weight, blood lipids, glucose metabolism, insulin sensitivity, and beta cell function were monitored.

Results

The body weight, serum cholesterol, and the Homeostasis Model Assessment ratio (insulin resistance index) were significantly reduced in JQ-R or metformin-treated mice, and the glucose tolerance was enhanced and insulin response was improved simultaneously. Moreover, both JQ-R and metformin could activate liver glycogen syntheses even under a relatively high glucose loading. Although glyconeogenesis was inhibited in the metformin treated mice, it was not observed in JQ-R treated mice. Similar to metformin, JQ-R could also improve the glucose infusion rate (GIR) in hyperglycemic clamp test. JQ-R was also shown to increase the levels of phosphorylated AMPKα and phosphorylated acetyl CoA carboxylase (ACC), similar to metformin.

Conclusion

JQ-R could reduce HFD-induced insulin resistance by regulating glucose and lipid metabolism, increasing insulin sensitivity through activating the AMPK signaling pathway, and subsequently improving β cell function. Therefore, JQ-R may offer an alternative in treating disorders associated with insulin resistance, such as prediabetes and T2DM.     

黄连可溶蛋白SDS-PAGE电泳鉴别

目的对雅连、峨眉野连、云连及6个不同产地的味连进行电泳鉴别。方法采用SDS-PAGE电泳方法。结果味连与雅连具有显著差异,味连与峨眉野连之间几乎无差异,不同产地的味连差异小,云连仅一条极弱的条带。结论SDS-PAGE电泳可以作为黄连质量控制的鉴别依据。     

雅连和味连对内毒素致小鼠死亡率的影响

目的:研究雅连和味连对内毒素所致小鼠死亡率的影响。方法:采用小鼠尾静脉注射脂多糖的方法,连续48h观察小鼠死亡情况。结果:味连小剂量组能明显延长小鼠受脂多糖攻击后的存活时间(P<0.01),雅连小剂量组和味连小剂量组均能降低脂多糖致小鼠死亡率,与模型组相比,差异有统计学意义(P<0.01,P<0.001)。结论:味连小剂量能够明显延长内毒素小鼠的生存时间,降低死亡率,其作用优于雅连。     

黄连中盐酸小檗碱的水提工艺研究

目的:优选黄连中盐酸小檗碱的最佳提取工艺条件。方法:采用正交试验设计,以盐酸小檗碱的含量为考察指标,考察加水倍量、提取次数和提取时间3因素对提取结果的影响。结果:盐酸小檗碱的最佳提取工艺为加药材12倍量的水回流提取1次,提取时间为1h。结论:该优化提取工艺稳定可行,重现性好,可用于黄连中小檗碱的提取。     

基于GLUT4转位的中药及活性成分改善胰岛素抵抗研究进展

对中药通过调控葡萄糖转运蛋白4(glucose transporter 4,GLUT4)转位而改善胰岛素抵抗(insulin resistance,IR)的研究进展进行总结.GLUT4转位研究对IR的机制研究以及药物作用靶点的选择都具有重要意义.本文主要从磷脂酰肌醇3-激酶/非典型蛋白激酶C/蛋白激酶B(PI3K/ PKC/ Akt),AMP活化蛋白激酶(AMP-activated protein kinase,AMPK)等关键途径,介绍各种中药活性成分、单方及复方通过调控GLUT4转位对IR的改善作用及机制,为防治IR中药的临床合理使用及新药开发提供参考.     

正交实验筛选黄连水提工艺

目的：优选黄连最佳酸水提取工艺。方法：采用正交试验法,以提取液中小檗碱含量为考察指标,对影响黄连提取工艺的因素进行了研究。结果：实验设计因素中回流次数影响最为显著。结论：黄连的最佳提取工艺为2％盐酸,12倍量水,回流3次,每次1．5h。     

应用模糊数学方法进行黄连引种适栽区划的研究

本文应用的模糊数学方法，选择与黄连生物学特性有较大关系的气候生态因子，对我国黄连引种栽培提出适栽区别，以减少黄连等药用植物的盲目引种。     

长期服用非典型抗精神病药对患者体重指数、血浆瘦素和胰岛素低抗的影响

 目的 探讨长期服用氯氮平、奥氮平及利培酮对精神分裂症患者体重指数、血浆瘦素和胰岛素水平及胰岛素抵抗的影响及其之间的关系。方法 74例符合中国精神障碍分类与诊断标准第三版（CCMD-3）的精神分裂症长期单一用药患者与首发精神分裂症患者分为对照组（15例）、氯氮平组（27例）、利培酮组（19例）及奥氮平组（13例）,测定并比较各组患者的血浆瘦素（leptin）、空腹胰岛素（FINS）水平及胰岛素抵抗指数（HOMA-IR）。结果 ①长期经氯氮平、奥氮平及利培酮治疗可致精神分裂症患者体重指数（BMI）显著增高（P<0.05）,且氯氮平与奥氮平的作用强度大于利培酮;②长期经氯氮平、奥氮平及利培酮治疗可致精神分裂症患者血浆瘦素水平显著增高（P<0.05）,影响程度：氯氮平> 奥氮平> 利培酮;③长期经氯氮平、奥氮平及利培酮治疗的精神分裂症患者空腹胰岛素及胰岛素抵抗指数水平较首发精神分裂症患者高,但组间无统计学差异;④相关性分析结果显示,精神分裂症患者体重指数与其血浆瘦素、空腹胰岛素水平及胰岛素抵抗指数均呈显著正相关（P<0.05）。结论 长期单一使用氯氮平、奥氮平及利培酮治疗,会导致精神分裂症患者体重增加,血浆瘦素、空腹胰岛素水平及胰岛素抵抗指数升高,且体重指数与血浆瘦素、空腹胰岛素水平及胰岛素抵抗指数均呈显著正相关,提示长期单一应用氯氮平、奥氮平及利培酮引起血浆瘦素和胰岛素抵抗指数增高是其所致代谢紊乱的重要原因之一。     

Umbelliferone Improves an Impaired Glucose and Lipid Metabolism in High‐Fat Diet/Streptozotocin‐Induced Type 2 Diabetic Rats

Umbelliferone (UMB) is a natural product that has several pharmacological effects including antihyperglycemic activity in diabetic rats. Thus, the objective of this study was to investigate the effect of UMB on insulin resistance and on the regulation of glucose and lipid metabolism in type 2 diabetic rats. Type 2 diabetes was induced in rats by feeding a high‐fat diet (45 kcal% fat) and a single dose of streptozotocin injection. After 8 weeks of treatment, UMB significantly reduced the elevated blood glucose levels and insulin resistance and increased the liver glycogen and serum adiponectin. Moreover, the serum lipid and the storages of triglyceride and non‐esterified fatty acid in liver tissue were reduced. From histological examination, the lipid droplets in liver tissue were clearly decreased, and the fat cell size in the fat tissue was smaller in diabetic rats treated with UMB. Interestingly, UMB increased fat cell adiponectin, plasma membrane glucose transporter 4 (GLUT4) and peroxisome proliferator‐activated receptor gamma (PPARγ), and liver PPARα protein expressions. Our findings demonstrate that UMB improves glucose and lipid metabolism in type 2 diabetes by stimulating the insulin secretion and the related mechanisms via stimulating expression of adiponectin, GLUT4, PPARγ, and PPARα‐protein expressions. Copyright © 2015 John Wiley & Sons, Ltd.     

芒果苷对HepG2细胞胰岛素抵抗的影响

目的探讨芒果苷在体外的降糖效果及对胰岛素传导关键信号蛋白磷酸化蛋白激酶B[p-AKT(Thr308)]、磷酸化糖原合成激酶3β[p-GSK-3β(Ser9)]、腺苷酸活化蛋白激酶(AMPKα)、葡萄糖转运蛋白2(GLUT2)表达的影响。方法培养HepG2细胞,用CCK-8法检测芒果苷对HepG2细胞增殖的影响;采用浓度为1×10^-6 mol/L的胰岛素刺激细胞36 h建立胰岛素抵抗模型;用葡萄糖检测试剂盒检测芒果苷对胰岛素抵抗HepG2细胞葡萄糖消耗的影响,用western blot法探讨其可能的作用机制。结果芒果苷浓度在>125μg/mL浓度后对HepG2细胞生长有抑制作用,故采用浓度为60、30、15μg/mL的芒果苷进行葡萄糖消耗实验,发现实验组与模型组比较,葡萄糖消耗量明显增加且具有显著性差异(P<0.01,P<0.05)。western blot结果表明,与正常组比较,模型组p-AKT(Thr308)、p-GSK-3β(Ser9)、AMPKα、GLUT2蛋白表达明显降低且存在显著性差异(P<0.05);与模型组比较,实验组蛋白表达均明显增加,且高剂量组均存在显著性差异(P<0.01,P<0.05),低剂量组除GLUT2蛋白表达具有显著性差异(P<0.05),其他蛋白虽有增加但无统计学意义。结论芒果苷对HepG2细胞胰岛素抵抗有改善作用,其作用机制可能是通过调控p-AKT(Thr308)、p-GSK-3β(Ser9)、AMPKα及GLUT2等蛋白的表达来改善2型糖尿病的胰岛素抵抗。     

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??OBJECTIVE To investigate the Polygonum capitatum's influences on the related indicators in db/db mice which is the obesity model of type 2 diabetes mellitus. METHODS Randomly dividing the mice into 5 groups:model group, rosiglitazone hydrochloride group, low-, moderate- and high-dose groups of Polygonum capitatum (5, 10, 20 g??kg^-1), make the db/m mice as blank control. Give the medicine for four weeks. The body weight, blood sugar were determined every week.At the end of fourth week, measuring the glucose tolerance and INS, IL-6 in serum. After all the mice were killed, testing the cholesterol and triglyceride in liver and skeletal muscle and then collecting the liver tissue for HE staining. At the meantime, the expression level of AMPK and GLUT4 in liver were detected by Q-PCR. RESULTS Polygonum capitatum can improve the body weight, blood sugar and glucose tolerance of db/db mice as well as the content of INS and IL-6 in serum, but increase the content of SOD and decrease the content of MDA in mice, furthermore, the cholesterol and triglyceride levels in the liver and skeletal muscle were also declined. HE staining showed that Polygonum capitatum could reduce the number of vacuoles in the liver of db/db mice, and make its shape more complete and ordered. What's more, raising the expression of AMPK and GLUT4 in the liver. CONCLUSION Polygonum capitatum can improve the condition of insulin resistance state, alleviate inflammation and advance the ability of db/db mice, which can also reduce the number of vacuoles in liver, and relieve the tissue lipid metabolic disorder. Meanwhile, Polygonum capitatum can promote the uptake of glucose in liver tissues, which is resulted from up-regulation of expression in hepatic AMPK and GLUT4 gene.     

壳聚糖絮凝沉淀工艺糖对维康浸膏剂疗效的影响

目的:考察壳聚糖对"糖维康(TWK)"浸膏剂絮凝效果,优选最佳沉淀工艺。研究壳聚糖糖维康浸膏剂(K-TWK)对治疗胰岛素抵抗(Insulin resistance,IR)大鼠血糖、血脂水平、胰岛素等的影响,探讨其除杂工艺的合理性。方法:以澄明度、总固型物收率为指标,进行制剂工艺考察;采用高脂高糖饲料喂养法复制IR大鼠的动物模型,测胰岛素、血糖、TG、LDL-C、TC及HDL-C;采用放射免疫分析方法检测血中胰岛素、瘦素、TNF-α及C-肽水平,评价K-TWK与TWK的生物等效型。结果:K-TWK与TWK均能提高IR大鼠对胰岛素的敏感性,降低肝指数和血脂水平(P<0.05,P<0.01),降低IR大鼠血清瘦素、TNF-α水平(P<0.01),升高C-肽水平(P<0.05);二者相比无显著差异。结论:K-TWK与TWK在治疗糖尿病方面,二者无显著差异(P>0.05),通过壳聚糖絮凝沉淀可有效降低TWK杂质而不影响其药理作用。     

基于AMPK信号通路探讨花旗泽仁对2型糖尿病糖、脂代谢作用的实验研究

目的:基于AMPK信号通路探讨花旗泽仁对2型糖尿病糖、脂代谢的作用机制。方法:db/m小鼠10只,为空白组;db/db小鼠40只,随机分为4组,每组10只,为模型组、阳性药组(0.2 g/kg)、花旗泽仁高剂量组(32 g/kg)、花旗泽仁低剂量组(16 g/kg)。给药6周后,检测各组小鼠空腹血糖(FPG)及血清中胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C),Western blot法检测各组小鼠肝脏中单磷酸腺苷酸活化蛋白激酶(AMPK)、促进葡萄糖转运体4(GLUT4)、瘦素(Leptin)、过氧化物酶体增殖活化受体-1a(PGC-1a)蛋白质相对表达量。结果:花旗泽仁可显著降低FPG及血清中TC、TG和HDL-C的含量,上调小鼠肝脏中AMPK、GLUT4、Leptin、PGC-1a蛋白质相对表达量。结论:花旗泽仁可显著改善db/db小鼠糖、脂代谢紊乱,其作用可能与调节肝脏中AMPK相关信号通路有关。     

血清芽管实验检测白假丝酵母菌的药物敏感性

目的了解泌尿生殖道感染白假丝酵母菌对黄连、黄芩、七叶一枝花和氟康唑注射液的敏感性,为临床有效控制其感染提供依据。方法采用白假丝酵母菌芽管药敏试验法,测定黄连、黄芩、七叶一枝花的50%乙醇提取物和氟康唑注射液对从泌尿生殖道感染标本中分离出的52株白假丝酵母菌芽管的最低抑制浓度。结果黄连、黄芩、七叶一枝花的50%乙醇提取物和氟康唑注射液,在一定的浓度下对泌尿生殖道感染白假丝酵母菌芽管有明显的抑制作用,其抑制芽管的最低浓度分别为1.25,5,2.5 mg/ml和0.125 mg/ml。结论白假丝酵母菌芽管药敏试验法,可为临床快速筛选敏感药物;黄连、黄芩、七叶一枝花的50%乙醇提取物和氟康唑注射液对白假丝酵母菌有较好的抗菌活性。     

黄芪黄酮与葛根黄酮配伍对肝脏糖脂代谢的影响

目的:明确黄芪黄酮与葛根黄酮配伍对糖尿病大鼠糖脂代谢的影响。方法:SPF级SD雄性大鼠66只,按血糖随机分为6组,11只/组。除正常组外,其他各组均按46 mg·kg-1予以1次性ip链脲佐菌素(STZ)诱导糖尿病模型。造模同日ig给药,黄芪黄酮组、葛根黄酮组、黄芪黄酮+葛根黄酮组分别给予0.039,0.036,0.075 g·kg-1;阳性药组给予金芪降糖片1.47 g·kg-1;正常组、模型组ig给予等量蒸馏水10 m L·kg-1。各组连续给药30 d。分别检测血糖、血清胆固醇(cholesterol,CHO)与甘油三酯(triglyceride,TG)含量,检测肝脏胰岛素(insulin,INS)、葡萄糖转运体4(glucose transporter 4,GLUT4)、脂联素(adiponectin,ADPN)及瘦素(leptin,LEP)浓度。运用单因素方差分析法分析两种药物的单独效应,析因设计方差分析法分析两种药物合用的交互效应。结果:与正常组比较,模型组大鼠血糖、血清CHO,TG均升高(P0.05),肝组织INS,GLUT4,ADPN及LEP浓度均下降(P0.05)。与模型组比较,黄芪黄酮组糖尿病大鼠血糖水平明显降低(P0.05),葛根黄酮组变化不明显,二药合用未呈现协同作用;黄芪黄酮组、葛根黄酮组CHO,TG含量均明显降低,但合用效果不如单味药;黄芪黄酮、葛根黄酮均能明显升高肝脏GLUT4水平(P0.05);二药对肝脏INS,GLUT4及LEP的影响,合用比单味药效果好,二药存在协同作用。结论:黄芪黄酮与葛根黄酮配伍能够协同调节INS,GLUT4及LEP水平而实现改善糖尿病糖脂代谢紊乱的作用。