Influence of berberine on protein tyrosine kinase of erythrocyte insulin receptors from type 2 diabetes mellitus

Objective： Bererine has been used to treat type 2 diabetes mellitus in Chinese traditional medicine because of its hypoglycemic effect. In this report, we compared the intrinsic tyrosine kinase activities of erythrocyte insulin receptors from type 2 diabetes mellitus with or without stimulation by berberine in vitro. Methods- Preparations containing insulin receptors were obtained from soluble human erythrocytes, and the insulin receptors were partially purified by affinity chromatography. The tyrosine kinase activity was measured by the exogenous substrate phosphorylation. Results： Both the membrane tyrosine kinase activity and the purified receptor tyrosine kinase activity from diabetics decreased significantly compared with those of normal individuals （reduced by 67.4% and 47.2%, respectively）. After incubation with berbefine, there is a statistical difference in the activity of membrane tyrosine kinase for diabetic patients （ a 150% increase）. Berefine had no effect on the tyrosine kinase activity of purified insulin receptors. Conclusion： We concluded from these results that berbefine was able to improve the insulin sensitivity by increasing the protein tyrosine kinase activity of membrane-bound insulin receptors from type 2 diabetes mellitus.     

不同品系黄连产量和质量的研究

目的考察和比较湖北黄连GAP基地选育的3个不同品系黄连(大花叶、无光叶、小花叶)的产量和质量。方法采用田间随机区组设计,连续3年采集3个品系黄连根茎样品,分别以烘干法测定其质量,用紫外分光光度法和高压液相色谱法分别测定其总生物碱和盐酸小檗碱的量。结果在考察的3个品系药材样本中,各品系三年生黄连产量没有显著差异(P>0.05),在四年生和五年生黄连中,大花叶和无光叶品系产量均明显高于小花叶品系(P<0.05);在三年生、四年生、五年生黄连中,各品系总生物碱和盐酸小檗碱的量无明显差异(P>0.05)。结论大花叶和无光叶品系的黄连产量高、质量合格,可作为黄连规范化种植的种子基原。     

反相高效液相色谱法测定复方小檗片中盐酸小檗碱的含量

目的采用反相高效液相色谱(RP-HPLC)法测定复方小檗片中盐酸小檗碱的含量。方法用Krom asil色谱柱(5μm,250 mm×4.6 mm),流动相∶乙腈-水-磷酸二氢钾-十二烷基磺酸钠(500 m l∶500 m l∶3.0 g∶1.5 g),流速:1 m l.m in-1,柱温:40℃,检测波长为345 nm。结果在0.021 2~0.233 2μg之间,峰面积与进样量的线性关系为Y=3 513 092.99X-15 938.76(r=0.999 9),平均回收率为97.22%。测得3批片剂中盐酸小檗碱的含量分别为0.772,0.748,0.749 mg/片。结论所建方法简便、准确、重复性好,可用于复方小檗片中盐酸小檗碱的含量测定。     

高效液相色谱法测定三黄片中盐酸小檗碱的含量

目的建立三黄片中盐酸小檗碱的含量测定方法。方法高效液相色谱法，色谱柱为Kromasil^TMC，s柱，每1000mL含磷酸二氢钾3．4g、十二烷基硫酸钠1，7g的乙腈-水（65：35）为流动相，流速1mL／min，检测波长345nm。结果盐酸小檗碱进样量线性范围是0．0357．0，357μg，r=0．9999，平均回收率为100．3％，RSD=0．77％（n=6）。结论该法操作简便、快捷、准确，可用于三黄片的质量控制。     

小檗碱对人胃癌细胞株BGC-823增殖的影响

用MTT法、克隆分析、流式细胞仪及免疫组织化学法 ,观察了小檗碱对人胃癌细胞株BGC 82 3增殖的影响 ,结果发现小檗碱对人胃癌细胞株BGC 82 3有抑制增殖作用 ,且与药物浓度呈正相关。其机理主要为使突变型 p5 3和 p2 1蛋白的表达下调 ,阻滞胃癌细胞进入细胞周期 ,使人胃癌细胞BGC 82 3的细胞周期阻滞于G0 /G1期。     

反相高效液相色谱法测定如意金黄贴膏中小檗碱的含量

 目的：黄柏在如意金黄贴膏中为君药，建立黄柏中小檗碱的定量方法。方法：采用反相高效液相色谱法测定小檗碱的含量。使用μ-BondapakTM C₁₈(3.9 mm×300 mm，10 μm)色谱柱，以乙腈-磷酸二氢钾-磷酸（300∶700∶1.50）为流动相，检测波长为350 nm。结果：此法线性关系良好，平均回收率为100.56%。结论：本方法精密度高，分离度良好，可用于如意金黄贴膏的质量控制。     

复方黄连素治疗腹泻型肠易激综合征疗效观察

目的探讨复方黄连素对肠易激综合征中的疗效。方法136例腹泻型肠易激综合征患者,随机分为两组,各68例。以复方黄连素、培菲康、匹维溴胺、多虑平治疗为治疗组,对照组中无复方黄连素,疗程2周。结果两组疗效比较差异有统计学意义,P<0.01。结论复方黄连素治疗腹泻型肠易激综合征效果良好,建议临床进一步推广。     

左金胃漂浮缓释片中小檗碱、巴马汀、吴茱萸碱和吴茱萸次碱的体外释放规律和机制研究

目的探讨左金胃漂浮缓释片体外多成分释放的规律和机制。方法采用转篮法以人工胃液为介质,以HPLC法测定左金胃漂浮缓释片中小檗碱、巴马汀、吴茱萸碱和吴茱萸次碱在8 h内的体外累积释放率,通过相似因子比较法、Higuchi方程,零级、一级释放方程等释放模型拟合法以及Peppas方程研究左金胃漂浮缓释片多成分释放的规律。结果左金胃漂浮缓释片中4种生物碱的平均释放曲线相互之间的相似因子均大于80%,均以Higuchi释放模型为最佳拟合模型,释放机制均为扩散协同骨架溶蚀作用。结论左金胃漂浮缓释片中生物碱类成分的释放具有均衡缓释性。     

RP-HPLC法测定中药材黄柏中小檗碱的含量

用反相高效液相色谱法,安定为内标,ZorbaxC₈柱,含30mmol/L。磷酸的乙腈-水(35:65)为流动相,于紫外346nm处测定了黄柏中小檗碱的含量。排除了巴马汀对小檗碱的干扰。平均回收率为101.7%,RSD为1.83%(n=20)。     

Oxygen activation by photoexcited protoberberinium alkaloids from Mahonia aquifolium

Protoberberinium salts, i.e. berberine (I), palmatine (II) and jatrorrhizine (III) prepared from Mahonia aquifolium (Pursh) Nutt. belong to isoquinoline alkaloids possessing interesting biological activity (e.g. antibacterial, antimalarial, antitumor). The characteristic UV/Vis absorption band maxima of I-III iodide salts were found in regions 350 and 425 nm in dimethylsulfoxide (DMSO) and ethanol solvents, and were only negligibly influenced by substitution changes on the C-2 and C-3 positions. The fluorescence intensity of protoberberinium salts monitored in ethanol solutions was significantly lowered by iodide counter-ions, and decreased in the order berberine > palmatine > jatrorrhizine. EPR spectroscopy supplied evidence of the formation of super-oxide anion radicals and singlet oxygen upon irradiation of berberine in oxygenated DMSO solvent. The photochemical generation of O(2) (.-) and (1)O(2) in DMSO solutions of palmatine and jatrorrhizine was substantially lower, and probably reflected the replacement of a photolabile methylenedioxy group at C-2 and C-3 positions in the berberine molecule by two methoxy groups in palmatine, and methoxyl (C-2) and hydroxyl (C-3) substitution in jatrorrhizine. Additionally, the powder EPR spectra of protoberberinium iodides I-III measured at 290 K revealed the presence of single-line EPR signals (g(eff) = 2.0044), which were attributed to hydroperoxidic structures produced by the autoxidation process. The photochemical reactions of protoberbenium salts producing reactive oxygen species after UVA excitation should be integrated in biological activity investigations, as well as in their applications in skin disorder treatment.