小檗碱通过影响糖原结构调节肝糖代谢研究

目的探讨小檗碱调节糖原代谢的机制及对糖原结构的影响。方法采用自发型糖尿病模型db/db小鼠作为疾病模型鼠,考察小檗碱对db/db小鼠血糖水平的影响;提取小鼠的肝糖原进行体积排阻色谱(SEC)和透射电镜(TEM)分析,考察小檗碱对db/db小鼠肝糖原的结构影响,并探讨其影响机制。结果小檗碱可以显著性降低db/db小鼠的空腹血糖,降低db/db小鼠血清胰岛素水平;改善db/db小鼠肝糖原的结构不稳定性;降低db/db小鼠肝组织中糖原磷酸化酶(GP)、糖原脱分支酶(GDBE)和环磷酸腺苷(cAMP)表达水平,降低血清胰高血糖素水平。结论小檗碱可以调控cAMP/GP信号通路,改善db/db小鼠的肝糖原结构,修复受损糖原结构,这可能是其调节肝糖代谢,改善糖尿病症状的机制之一。     

Berberine ameliorates non-steroidal anti-inflammatory drugs-induced intestinal injury by the repair of enteric nervous system

The study was to detect the role of GDNF, PGP9.5 (a neuronal marker), and GFAP (EGCs’ marker) in the mechanism of non-steroidal anti-inflammatory drugs (NSAIDs) related to intestinal injury and to clarify the protective effect of berberine in the treatment of NSAID-induced small intestinal disease. Forty male SD rats were divided randomly into five groups (A–E): Group A: control group; Group B: model group received diclofenac sodium 7.5 mg/(kg*day) for 5 days; Group C–E: berberine low, medium and high dose groups were treated by 7.5 mg/(kg*day) diclofenac sodium for 5 days then received berberine 25 mg/(kg*day), 50 mg/(kg*day), and 75 mg/(kg*day), respectively, between the sixth and eighth day. Intestinal mucosa was taken on the ninth day to observe the general, histological injuries, and to measure the intestinal epithelial thickness. Then, immunohistochemistry was performed to detect the expression of PGP9.5 and GFAP, and Western blot was performed to detect GDNF expression. The histological score and the general score in the model group were, respectively, 5.75 ± 1.04 and 4.83 ± 0.92. Scores in berberine medium and high berberine group were lower compared with the model group (P < 0.05). The intestinal epithelial thickness in the model group was lower than in the control group and the berberine groups (P < 0.05). PGP9.5, GFAP, and GDNF content in the model group and the three berberine groups were significantly lower than in the control groups (P < 0.05). PGP9.5, GFAP, and GDNF content in the control group and the three berberine groups were higher compared with the model groups (P < 0.05). Berberine can protect the intestinal mucosa of NSAID users, and the mechanism is associated with the reparation of the enteric nervous system via upregulating the expression of PGP9.5, GFAP, and GDNF.     

Berberine diminishes cancer cell PD-L1 expression and facilitates antitumor immunity via inhibiting the deubiquitination activity of CSN5

Programmed cell death-1 (PD-1)/programmed cell death ligand-1 (PD-L1) blocking therapy has become a major pillar of cancer immunotherapy. Compared with antibodies targeting, small-molecule checkpoint inhibitors which have favorable pharmacokinetics are urgently needed. Here we identified berberine (BBR), a proven anti-inflammation drug, as a negative regulator of PD-L1 from a set of traditional Chinese medicine (TCM) chemical monomers. BBR enhanced the sensitivity of tumour cells to co-cultured T-cells by decreasing the level of PD-L1 in cancer cells. In addition, BBR exerted its antitumor effect in Lewis tumor xenograft mice through enhancing tumor-infiltrating T-cell immunity and attenuating the activation of immunosuppressive myeloid-derived suppressor cells (MDSCs) and regulatory T-cells (Tregs). BBR triggered PD-L1 degradation through ubiquitin (Ub)/proteasome-dependent pathway. Remarkably, BBR selectively bound to the glutamic acid 76 of constitutive photomorphogenic-9 signalosome 5 (CSN5) and inhibited PD-1/PD-L1 axis through its deubiquitination activity, resulting in ubiquitination and degradation of PD-L1. Our data reveals a previously unrecognized antitumor mechanism of BBR, suggesting BBR is small-molecule immune checkpoint inhibitor for cancer treatment.     

Renal vectorial transport of berberine mediated by organic cation transporter 2 (OCT2) and multidrug and toxin extrusion proteins 1 (MATE1) in rats

Berberine, a well‐known plant alkaloid derived from Rhizoma coptidis, has potential applications as a therapeutic drug for diabetic nephropathy. However, the transporter‐mediated renal transport of berberine remains largely unclear. This study aimed to investigate the renal transport mechanism of berberine using transfected cells, kidney slices and animal experiments. In Madin‐Darby canine kidney (MDCK) cells stably expressing rat OCT2 (MDCK‐rOCT2) and kidney slices, saturable and non‐saturable uptake of berberine was observed, and corticosterone could inhibit the uptake of berberine, with IC₅₀ values of 0.1 μm and 147.9 μm , respectively. In double‐transfected cells, the cellular accumulation of berberine into MDCK‐rOCT2 and MDCK‐rOCT2‐rMATE1 (MDCK cells stably expressing rOCT2 and rMATE1) cells was significantly higher than the uptake into MDCK cells. Meanwhile, berberine transcellular transport was considerably higher in double‐transfected MDCK‐rOCT2‐rMATE1 cells than in MDCK and MDCK‐rOCT2 cells. Corticosterone for MDCK‐rMATE1 and MDCK‐MDR1 and pyrimethamine for MDCK‐rMATE1 at high concentrations could inhibit the efflux of berberine. In animal experiments, compared with the berberine alone group, the cumulative urinary excretion of berberine significantly decreased in the corticosterone or pyrimethamine pretreatment groups. In the rat kidney, pyrimethamine increased, and a low dose of corticosterone (5 mg/kg) decreased, the berberine concentration. However, there was no apparent change in the renal concentration of berberine in rats pretreated with corticosterone (10 or 20 mg/kg). Thus, berberine is not only a substrate of OCT2 and P‐glycoprotein, but is also a substrate of MATE1. Both OCT2 and MATE1 mediate the renal vectorial transport of berberine.     

基于网络搜索快速鉴定抗风湿类中成药和保健食品中非法添加的非那西丁及小檗碱

目的 鉴定抗风湿类中成药和保健食品中非法添加的2个未知成分。方法 采用高效液相色谱-二极管阵列检测器联用（HPLC-DAD）技术进行抗风湿类中成药和保健食品非法添加筛查时发现2个可疑色谱峰,采用UPLC-MS/MS技术获得其准分子离子和二级质谱图,然后通过网络搜索分析初步确定可疑成分,最后与对照品比较检测,最终确定2个非法添加化合物的结构。结果 在抗风湿类保健食品中检出盐酸小檗碱,中成药中检出非那西丁。结论 这2个化合物不在现有抗风湿类中成药和保健食品检验标准13种目标化合物范围内,容易逃脱标准监管范围。     

小蘖碱对高脂血症鼠脂联素的调控作用及其对血脂指标和斑块形成的影响

目的 研究小蘖碱对高脂血症鼠脂联素的调控作用及其对血脂指标和斑块形成的影响。 方法 在40只Wistar雄性鼠随机选取30只并喂养高脂饲料饮食进行高血脂(HLP)造模,对照组(n=10)则喂养正常食物并自由摄取水。10周后,将HLP模型随机分为3组: HLP组(n=10)、HLP+LBB组(n=10)和HLP+HBB组(n=10)。对HLP+LBB组鼠进行低剂量小檗碱(berberine, BB)灌胃(150 mg/(kg·d)), HLP+HBB组高剂量小檗碱灌胃(300 mg/(kg·d)),对照组和HLP组则以生理盐水代替。连续喂养10周后测定并比较4组鼠血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、血清总脂联素和高分子量脂联素含量,以及心肌梗死斑块面积。 结果 造模后,与对照组相比,HLP组大鼠TG、TC和LDL-C显著升高,HDL-C水平显著下降(P<0.05)。提示HLP大鼠造模成功。与HLP组相比,HLP+LBB组、HLP+HBB组TG、TC和LDL-C显著降低(P<0.001),HLP+LBB组HDL-C水平显著升高(P<0.05)。四组大鼠血清总脂联素含量差异无统计学意义(P>0.05),但HLP组中高分子量脂联素与总脂联素的比值显著低于对照组, 而HLP+HBB组显著高于HLP组(P<0.05)。与对照组比较,AdipoR1在HLP组中表达下调(P<0.05), 而HLP+HBB组显著高于HLP组,差异有统计学意义(P<0.05)。AdipoR2在HLP组中的表达无明显变化,但HLP+HBB组显著高于对照组(P<0.05)。HLP+HBB组梗塞面积[(11.4±1.0)%)]显著低于HLP组[(52.2±0.9)%]和HLP+LBB组[(33.1±1.1)%](均P<0.05)。 结论 小蘖碱可对抗高脂血症鼠血中脂联素的下调,这一干预可能在血脂指标和斑块形成方面起到改善作用。     

小檗碱治疗代谢性疾病抗炎作用的研究进展

慢性非可控性炎症在代谢性疾病如糖尿病、非酒精性脂肪性肝病（NAFLD）和动脉粥样硬化的发生和发展过程中发挥关键作用。天然产物小檗碱对代谢性疾病有良好的改善作用并对其伴随的慢性炎症有明显的抑制作用。研究显示小檗碱主要通过调控AMP依赖的蛋白激酶（AMPK）、核转录因子-κB（NF-κB）和肠道菌群来发挥抗炎活性。对小檗碱抗炎作用的深入研究将为其应用于临床治疗代谢性疾病提供进一步科学依据。     

蛋白质在膜表面的吸附特性及蛋白质存在的溶液环境对小檗碱膜透过行为的影响研究

为探索蛋白质在膜表面的吸附特性以及蛋白质存在的溶液环境对小檗碱膜透过行为的影响,以蛋白质、小檗碱为研究对象,配制模拟溶液,通过低场核磁共振技术、静态吸附实验以及膜分离实验探究蛋白质与陶瓷膜、小檗碱之间可能存在的相互作用,以蛋白质静态吸附量、膜过程相对通量、蛋白质截留率、小檗碱透过率及两者的吸附率为评价指标,同时测定膜过程中膜污染阻力分布、过膜前后粒径分布变化及膜表面电镜扫描(SEM),研究蛋白质在陶瓷膜上的吸附特性及对小檗碱膜过程的影响。结果表明,陶瓷膜对蛋白质具有吸附作用,且吸附模型符合Langmuir吸附模型,在模拟溶液的膜分离过程中,蛋白质是引起膜污染的主要因素,但在1 g·L~(-1)蛋白质的浓度下,蛋白质的存在对小檗碱的膜过程没有显著影响。     

黄连素的药理作用及机制研究进展

黄连素是从黄连、黄柏等植物中提取出来的一种生物碱,其具有显著的抗菌作用,用来治疗细菌性胃肠炎、痢疾等消化道疾病已有很长的历史。近年来,黄连素的其他药理作用逐渐被发现。研究结果表明,黄连素对心血管系统、神经系统、内分泌系统等均有明显影响,尤其是降血脂作用对防治动脉粥样硬化效果良好。黄连素还能增加胰岛素的敏感性来治疗糖尿病。黄连素被发现有明显的抗炎抗癌作用。黄连素能杀伤肿瘤细胞抑制肿瘤细胞增殖和诱导肿瘤细胞的凋亡。黄连素还能影响钙离子、钾离子等离子通道来调节电解质平衡。由于黄连素的抗炎抗氧化作用,可用于治疗消化性溃疡。本文就黄连素的作用及其机制进行了详细的总结,包括黄连素对心血管系统、神经系统、内分泌系统的影响以及抗炎、抗菌、抗癌等药理作用,还有对离子通道、周期蛋白、细胞外基质的影响。有关黄连素的作用及机制越来越引人关注,尤其是抗癌活性,表明对含有黄连素这一活性成分的中药及方剂的研究将会有新的进展。总结黄连素的作用及其机制,将有助于提高黄连素的临床应用及新药研发。     

黄连饮片标准汤剂的制备及质量标准分析

目的:制备黄连饮片标准汤剂并建立其质量标准,为黄连配方颗粒的制备及其质量标准的建立提供参考。方法:根据中药饮片标准汤剂制备原则制备黄连饮片标准汤剂,计算表小檗碱、盐酸黄连碱、盐酸巴马汀和盐酸小檗碱的转移率和饮片出膏率,建立黄连饮片标准汤剂的质量标准。流动相乙腈-0.05 mol·L~(-1)磷酸二氢钾溶液(50∶50)(每100 m L中加十二烷基硫酸钠0.4 g,以磷酸调节pH 3.0),柱温30℃,流速1 mL·min~(-1),检测波长225 nm。结果:黄连饮片中表小檗碱、盐酸黄连碱、盐酸巴马汀和盐酸小檗碱的转移率范围分别为79.3%~111.9%,54.6%~76.2%,45.7%~70.7%和43.5%~64.4%,出膏率范围17.1%~22.3%,14批黄连饮片标准汤剂特征图谱中有7个共有峰,其相似度均0.999。结论:该研究建立的质量评价方法精密度和重复性良好,指纹图谱相似度高,适用于黄连饮片标准汤剂的质量评价。