Berberine and its structural analogs have differing effects on functional profiles of individual gut microbiomes

ABSTRACT

The understanding of the effects of compounds on the gut microbiome is limited. In particular, it is unclear whether structurally similar compounds would have similar or distinct effects on the gut microbiome. Here, we selected berberine (BBR), an isoquinoline quaternary alkaloid, and 16 structural analogs and evaluated their effects on seven individual gut microbiomes cultured in vitro. The responses of the individual microbiomes were evaluated by metaproteomic profiles and by assessing butyrate production. We show that both interindividual differences and compound treatments significantly contributed to the variance of metaproteomic profiles. BBR and eight analogs led to changes in proteins involved in microbial defense and stress responses and enrichment of proteins from Verrucomicrobia, Proteobacteria, and Bacteroidetes phyla. It also led to a decrease in proteins from the Firmicutes phylum and its Clostridiales order which correlated to decrease proteins involved in the butyrate production pathway and butyrate concentration. Three of the compounds, sanguinarine, chelerythrine, and ethoxysanguinarine, activated bacterial protective mechanisms, enriched Proteobacteria, increased opacity proteins, and markedly reduced butyrate production. Dihydroberberine had a similar function to BBR in enriching the Akkermansia genus. In addition, it showed less overall adverse impacts on the functionality of the gut microbiome, including a better maintenance of the butyrate level. Our study shows that ex vivo microbiome assay can assess differential regulating effects of compounds with subtle differences and reveals that compound analogs can have distinct effects on the microbiome.     

HPLC测定消炎栓中黄芩苷、盐酸小檗碱的含量

目的 采用高效液相色谱法同时测定消炎栓中黄芩苷和盐酸小檗碱的含量,以提高其质量标准.方法 Waters SymmetryShield-C18柱(4.6 mm× 250 mm,5 μm)为固定相,以乙腈-0.4％磷酸溶液进行梯度洗脱,流速为1.0 mL/min,检测波长为264nm.结果 黄芩苷、盐酸小檗碱的线性范围分别...     

高效液相色谱法测定二妙丸中盐酸小檗碱的含量

目的建立二妙丸中盐酸小檗碱的含量测定方法。方法采用高效液相色谱法,DiamonsilODSlC18柱,以乙腈-0．1％磷酸溶液（50：50,每100mL加十二烷基磺酸钠0．1g）为流动相,流速为1．0mL／min,检测波长265nm。结果盐酸小檗碱在0．1386～2．772μg范围内呈良好线性,回归方程为,：2572．354X＋79．851,r=0．9997。平均加样回收率为98．67％,RSD=1．26％（n=6）。结论方法简便、准确,专属性强,测定结果重复性好,为二妙丸中盐酸小檗碱的定量分析提供了科学有效的方法。     

HPLC测定岩黄连生物总碱中脱氢卡维丁、盐酸巴马汀和盐酸小檗碱

目的:建立高效液相色谱法测定岩黄连生物总碱中脱氢卡维丁、盐酸巴马汀及盐酸小檗碱含量的方法,为完善岩黄连生物总碱的质量评价提供依据。方法:采用高效液相色谱法,以Agilent XDB C18(4.6 mm×150 mm,5μm)为色谱柱,以乙腈-0.01 mol.L-1磷酸二氢钾水溶液(18.5∶81.5)为流动相,流速1.0 mL.min-1,柱温30℃,检测波长347 nm。结果:脱氢卡维丁进样量在15.375～92.25 mg.L-1(r=0.999 9)呈良好的线性关系,平均加样回收率96.43%,RSD 1.13%;盐酸巴马汀进样量在14.91～89.46 g.L-1(r=0.999 9)呈良好的线性关系,平均加样回收率96.28%,RSD 1.22%;盐酸小檗碱进样量在3.525～28.2 mg.L-1(r=0.999 7)呈良好的线性关系,平均加样回收率为103.02%,RSD 1.35%。结论:所建立的方法简便、准确、重复性好,可以用于控制岩黄连生物总碱的质量。     

高效毛细管电泳法测定三棵针中3种生物碱的含量

目的:建立毛细管区带电泳同时测定不同产地三棵针中盐酸小檗碱、盐酸药根碱、盐酸巴马汀含量的方法。方法:空心石英毛细管(75 cm×50μm)柱,以50 mmol.L-1硼砂-甲醇(2∶1)的溶液为电泳介质,运行电压25 kV,柱温15℃,检测波长230 nm。结果:盐酸小檗碱、盐酸药根碱及盐酸巴马汀分别在0.816 8～32.672 0 mg.L-1(r2=0.999 4),0.852 0～8.520 0 mg.L-1(r2=0.999 5),0.815 2～8.152 0 mg.L-1(r2=0.999 6)线性关系良好;其平均加样回收率分别为103.8%(RSD 1.14%),100.7%(RSD 3.46%),100.2%(RSD 2.87%)。结论:实验方法快速简便,准确可靠,适用于三棵针等药材中盐酸小檗碱、盐酸巴马汀及盐酸药根碱的含量测定。     

喷雾干燥条件对复方黄连干粉质量的影响

目的:通过实验优选出复方黄连干粉的最佳喷雾干燥制备条件。方法:以复方黄连干粉含水量及盐酸小檗碱损失率为指标,考察不同进风温度,喷速对复方黄连干粉质量的影响。结果:优选出最佳喷雾干燥条件,进风温度170～190℃,喷速3 500 mL.h-1。结论:采用上述喷雾干燥条件制备复方黄连干粉是可行的。     

微量热法对小檗碱类生物碱抑菌作用的量效关系研究

目的 测定黄连中的3种生物碱小檗碱、药根碱、巴马汀对痢疾杆菌生长代谢的影响,从热力学角度研究小檗碱类生物碱抑菌作用的量效关系。方法 以微量热法测定小檗碱类生物碱作用于痢疾杆菌生长代谢的热谱曲线,并得出相应的热动力学参数,如生长速率常数、产热功率、最大热功率出现时间、抑制率、传代时间等,根据热动力学模型,拟合了3种生物碱产热功率-质量浓度关系,并进行初步的量效关系分析。结果 小檗碱、药根碱、巴马汀对痢疾杆菌的生长代谢均有不同程度的抑制作用,在 0.08～1.20 mg/mL 内随着质量浓度的增加,最大热功率出现时间和传代时间延长,生长速率常数降低和最大产热功率降低,抑制率增加,且与药物质量浓度呈现一定的量效关系。结论 3种生物碱对痢疾杆菌的抑菌活性的强度顺序为小檗碱 ＞ 巴马汀 ＞ 药根碱,从热力学角度研究抑菌类中药的量效关系具有定量、灵敏、快速等优势。     

Advance of studies on anti-atherosclerosis mechanism of berberine

Coptis Chinensis is a traditional Chinese medicine herb that has the effect of clearing heat and drying dampness,purging fire to eliminate toxin.Berberine is the main alkaloid of Coptis Chinensis,and recent researches showed that berberine had the effect of anti-atherosclerosis.This paper reviewed the anti-atherosclerosis mechanism of berberine,which may be related to regulating lipids,anti-inflammation, decompression,reducing blood sugar,and inhibiting vascular smooth muscle cell proliferation.     

清胃黄连丸快速TLC鉴别与小檗碱新荧光扫描定量

目的建立清胃黄连丸的快速TLC鉴别及小檗碱的高灵敏度荧光扫描定量方法。方法采用同一供试品溶液在2块薄层板上鉴别黄芩、牡丹皮、连翘、栀子和甘草,用荧光扫描法测定了清胃黄连丸中盐酸小檗碱的含量。结果盐酸小檗碱点样量在0.019 68～0.098 4 g内呈良好的线性关系。盐酸小檗碱平均回收率为99.41%,RSD为1.34%(n=9)。结论TLC鉴别斑点清晰,阴性无干扰。清胃黄连丸的快速TLC鉴别与小檗碱新荧光扫描定量,简便、快捷、准确、实用,有效提高了传统丸剂的质量检测速度。     

黄癸素对人口腔上皮样癌KB细胞株及化学诱导性仓鼠颊癌作用的研究

目的研究黄癸素对体外培养人口腔上皮样癌KB细胞及化学诱导仓鼠的口腔癌变抑制作用。方法 MTT法检测黄癸素对KB肿瘤细胞株的影响,计算24,48,72h的IC50。采用8周龄仓鼠,建立二甲基苯丙蒽(DMBA)诱发的颊囊癌模型,用黄癸素进行干预。结果黄癸素对KB细胞有抑制作用,24,48,72h的IC50分别为(12.74±0.13),(4.56±0.22),(2.88±0.11)mg/L;对KB细胞生长呈浓度、时间依赖性抑制作用。DMBA诱导的仓鼠在36,54,72mg/kg的黄癸素作用下,颊黏膜不典型增生与癌变的发生率有不同程度的降低,其中54,72mg/kg剂量组的病变总发生率分别为66.6%和53.3%,比模型组(86.6%)分别降低了20.0%和33.3%(P<0.01)。结论黄癸素能显著的抑制KB细胞生长,对化学物诱导的仓鼠口腔癌变也有抑制作用。