黄山药总皂甙肠内菌代谢及代谢产物吸收的研究

目的：本离体和整体观察人和大鼠肠内菌对黄山药总皂苷（DX）的代谢作用及整体给予DX后吸收入血的有效成分，方法：用薄层色谱（TLC）及电喷雾质谱（ESI－MS）法检测粪中DX及其代谢产物。整体给予大鼠灌服DX900mg/kg，于给药后不同时间采集尿及血清样品，用ESI－MS检测吸收入血成分。结果：DX容易被人和大鼠消化道菌群代谢，随着代谢时间的延长，出现了各种甾体皂苷的降解产物及终产物薯蓣皂苷元（Dio)。整体实验表明，在大鼠血及尿中均发现分子量为415．3的代谢产物，经ESIMS二级质谱分析，上述分子量的化合物为Dio。结论：DX可被人和大鼠肠内菌代谢，DX经口服后Dio被吸收入血。     

黄山药总皂苷肠内菌代谢及代谢产物吸收的研究

目的 :本文离体和整体观察人和大鼠肠内菌对黄山药总皂苷(DX)的代谢作用及整体给予DX后吸收入血的有效成分。方法 :用薄层色谱(TLC)及电喷雾质谱(ESI -MS)法检测粪中DX及其代谢产物。整体给予大鼠灌服DX900mg/kg ,于给药后不同时间采集尿及血清样品 ,用ESI -MS检测吸收入血成分。结果 :DX容易被人和大鼠消化道菌群代谢 ,随着代谢时间的延长 ,出现了各种甾体皂苷的降解产物及终产物薯蓣皂苷元 (Dio)。整体实验表明 ,在大鼠血及尿中均发现分子量为415 3的代谢产物 ,经ESI -MS二级质谱分析 ,上述分子量的化合物为Dio。结论 :DX可被人和大鼠肠内菌代谢 ,DX经口服后Dio被吸收入血。     

大鼠尿中人参皂苷Rd及其代谢物的LC-MS研究

目的探讨人参皂苷Rd在大鼠体内的代谢产物及转化途径。方法选择SD大鼠6只，单剂量口服和静脉给予人参皂苷Rd，分段收集给药前和给药后0～24 h尿样，将尿样分时段合并后采用旋转薄膜蒸发浓缩，以固相萃取小柱纯化处理，采用高效液相色谱-串联飞行时间质谱进行检测。结果通过比较给药前后的TOF总离子流图，对尿中推测的代谢物和标准物质的出峰时间及相关化合物选择离子扫描二级质谱图进行了比较分析，结果在尿中发现了7种代谢产物，系统分析了这些代谢产物的代谢转化规律及可能结构。结论大鼠尿中人参皂苷Rd的主要转化途径为氧化、水解、结合及异构化代谢反应。     

大鼠尿中黄芩汤多成分及其代谢物的分析比较

目的：通过大鼠尿中黄芩汤多成分及其代谢物的分析，比较复方和单味药中相应成分的代谢差异。方法：建立高效液相梯度洗脱的多成分分析方法，对口服黄芩汤及其各单味药后的大鼠尿中多种成分的排泄进行比较研究。结果：从尿中排泄物达峰时间（Tmax）比较，黄芩汤中的多种成分可以分为三种，一是快速排泄的成分，如芍药苷（PF）、甘草苷（LG）（Tmax为4h左右）；二是中等速度排泄的成分，如黄芩苷（BG）、汉黄芩苷（WG）、千层纸素A苷（OG）、黄芩素（B）和甘草酸（GL）（Tmax为8h-12h）；三是延迟排泄的成分，如汉黄芩素（W）、千层纸素A（O）、芍药苷代谢素I（PM-I）和甘草次酸（GA）（Tmax〉12h）。延迟排泄的成分都是代谢产物型化合物，经肠道内细菌作用，代谢转化后再吸收进入体内。在单味药中WG、OG、W、O、PM-I、LG和甘草素（L）的排泄总量和总排泄率高于在黄芩汤复方中。其中W、O、LG和L具有显著性差异，尤其是LG和L，其总排泄量是复方中的3倍左右。排泄速率在复方和单味药中也有差别，其中BG、WG、OG、B和GL的Tmax在复方中为8h，在单味药中缩短为4h，结论：黄芩汤大多数成分及代谢产物经尿的排泄在复方和在单味药中具有明显的差别，复方成分的代谢排泄较缓慢。     

穿龙薯蓣总皂苷中甾体皂苷的分离与鉴定

目的 研究穿龙薯蓣总皂苷中水溶性甾体皂苷，寻找新的活性化合物。方法 用硅胶柱色谱、薄层色谱及高效液相色谱等进行分离，通过酸水解、理化常数和波谱学分析（IR，NMR，MS，HMQC，HMBC）鉴定化合物结构。结果 从穿龙薯蓣总皂苷中分得2个甾体皂苷（1个水难溶性皂苷和1个水溶性皂苷），其化学结构分别鉴定为薯蓣皂苷元－3－O－{α-L-鼠李糖（1→2）－[β-D-葡萄糖（1→3）]}-β－D－葡萄糖皂苷（I），薯蓣皂苷元－3－O－[α－L－鼠李糖（1→3）－α-L-鼠李糖（1→4）－α－L－鼠李糖（1→4）]-β－D－葡萄糖皂苷（Ⅱ）。结论 Ⅱ为首次从穿龙薯蓣中分离得到的新化合物，命名为穿龙薯蓣皂苷Dc。     

淡紫青霉ACCC31890对纤细薯蓣皂苷的生物转化

摘 要 目的：研究淡紫青霉菌对纤细薯蓣皂苷的生物转化,分离鉴定转化产物并进行药理活性探讨。方法: 使用硅胶柱色谱和半制备液相色谱分离转化产物,依据质谱(MS）、核磁共振(NMR)波谱分析进行结构鉴定,并对产物进行了体外抗炎活性的研究。结果: 从转化粗提物中分离得到三个依次脱糖基产物,分别为5R-spirost-5-ene-3-ol-O-β-D-Glucopyranoside (1→3)-β-D-Glucopyranosyl（1）,延龄草苷（2）和薯蓣皂苷元（3）,转化率分别为1%,1%,45%;体外抗炎活性研究表明三种转化产物对LPS刺激巨噬细胞分泌NO,IL-6和MCP-1具有不同程度的抑制作用,其抗炎活性随着糖链的逐步水解有所增加,以终产物薯蓣皂苷元的抗炎活性最强。结论:淡紫青霉菌转化纤细薯蓣皂苷得到薯蓣皂苷元的产率较高,糖基的数量对甾体皂苷类化合物抗炎活性有着一定的影响。     

LC-MS/MS法研究1-[1-(6-甲氧基-2-萘基)乙基]-2-(4-硝基苄基)-6,7-二甲氧基-1,2,3,4-四氢异喹啉氢溴酸盐在大鼠体内的代谢产物

采用液相色谱-串联质谱法对大鼠灌胃1-［1-(6-甲氧基-2-萘基)乙基］-2-(4-硝基苄基)-6,7-二甲氧基-1,2,3,4-四氢异喹啉氢溴酸盐(编号P91024)后粪便、尿液、胆汁和血浆中的主要代谢产物进行研究。通过比较给药样品和空白样品的全扫描总离子流色谱和选择离子扫描色谱图差别寻找I相代谢产物；根据其一级和二级质谱图，确定I相代谢产物的分子结构。完全提取I相代谢产物后的样品溶液，再用葡糖醛酸酶酶解，得II相结合物的苷元部分，采用与I相代谢产物鉴定同样方法寻找和鉴定II相代谢产物苷元的结构，进而确证II相代谢产物的分子结构。从大鼠粪便中鉴定出P91024的2个I相代谢物，从胆汁中鉴定出1个I相和5个II相代谢产物，从尿液中鉴定出1个I相和3个II相代谢产物，从血浆中鉴定出4个I相和1个II相代谢产物；并分别分析推测出它们的结构。P91024在大鼠体内被代谢转化为多种产物，利用LC-MS/MS可以快速寻找和鉴定。     

基于 UPLC-Orbitrap-HRMS技术鉴定当归-川芎药对化学成分及大鼠体内入血成分

目的 鉴定当归-川芎药对化学成分及大鼠ig后的入血原型成分和代谢产物。方法 基于超高效液相色谱-静电场轨道阱-高分辨率质谱技术（UPLC-Orbitrap-HRMS）,结合对照品图谱、自建数据库、相关文献信息及Compound Discoverer3.3、Xcalibur qual browser 4.3等软件对当归-川芎药对的化学成分进行分析鉴定;大鼠经ig给药后制得含药血清,通过比对含药血清与对照组血清,鉴定入血原型成分和代谢产物。结果 共在当归-川芎药对中鉴定出69个化学成分,主要包括苯酞类24个、有机酸类24个、氨基酸类8个、含氮类7个、香豆素类4个、木脂素类1个、黄酮类1个;在含药血清中共鉴定出30个入血原型成分,包括苯酞类15个、有机酚酸类13个、香豆素类2个;鉴定出45个代谢产物,包括32个苯酞类代谢产物,13个有机酚酸类代谢产物。结论 明确了当归-川芎药对的化学成分及入血原型成分和代谢产物。     

离体大鼠肠道菌群对野漆树苷的代谢研究

摘 要 目的：研究大鼠肠道菌群对野漆树苷的体外代谢转化作用。方法: 将野漆树苷与离体大鼠肠道菌群的孵育液在37 ℃厌氧条件下孵育,不同时间点取样经乙酸乙酯萃取后,采用HPLC-MS/MS对代谢产物进行定性和定量分析。结果: 孵育4 h后野漆树苷已被肠道菌代谢完全,在代谢组的孵育液中发现了芹菜素和柚皮素两种代谢产物,以芹菜素为主,并且在4 h达到较高浓度,随后缓慢降解,24 h后消失。结论:在离体条件下,大鼠肠道菌群对野漆树苷具有显著的代谢转化作用,提示代谢产物为其生物活性成分。     

LC-MS/MS鉴定木兰花碱在大鼠体内外主要代谢产物

目的 研究木兰花碱在大鼠体内外的主要代谢产物及代谢途径。方法 SD大鼠ig木兰花碱（50 mg/kg）,收集0~24 h尿液和粪便,0~6 h胆汁以及1、2、4、6、8 h血浆;体外代谢采用肝微粒体温孵系统和肠菌培养液。利用LC-MS/MS对生物样品中的原型药及代谢产物进行鉴定。采用Agilent TC-C₁₈色谱柱（150 mm×4.6 mm,5 μm）,以乙腈-0.1%甲酸水溶液为流动相梯度洗脱,体积流量1.0 mL/min,柱温30℃。质谱采用电喷雾电离源（ESI）,正离子采集模式;扫描范围m/z100~1 000。根据药物体内代谢规则,结合木兰花碱的色谱保留时间和多级质谱碎片离子特征,推测其代谢产物的结构。结果 给药后生物样品中共鉴定出12个代谢产物,其中Ⅰ相代谢产物8个,Ⅱ相代谢产物4个。主要的代谢途径为羟基化、去甲基化、脱氢作用、酮基化、葡萄糖化、葡萄糖醛酸化及硫酸酯化。结论 木兰花碱在体内可发生Ⅰ相和Ⅱ相代谢,肠道菌群和肝药酶可催化木兰花碱发生Ⅰ相代谢转化,Ⅱ相代谢存在于肠道以外部位,最有可能的部位是肝脏。     

柚皮苷的人肠内菌生物转化(英文)

柚皮苷(1)是酸橙中含量最高的二氢黄酮苷类化合物, 将其与人肠内菌丛共温孵, 从温孵物中通过色谱方法得到原形化合物1和四个转化产物 (2-5), 根据谱学数据确定它们分别为柚皮苷-6"-乙酰物(2)、柚皮素(3)、根皮酸(4)和间苯三酚(5)。在1的糖苷链葡萄糖基6位能够特异性地发生乙酰化作用。1口服难以吸收。本文结果为阐明其生物利用度低提供了实验依据, 推测表现其生物活性的物质基础主要是其肠内菌生物转化产物3。肠内菌丛所致1的生物转化的继发结果,是它的转化产物吸收进入体循环发挥生物学作用, 此结果为进一步或再评价其体内过程、作用或毒性及其生物利用度提供了有用的信息。     

刺五加苷B、苷E在体外大鼠肠道菌群中的代谢

目的：观察体外大鼠肠道菌群对刺五加苷B、苷E的代谢。方法：收集大鼠新鲜粪便在厌氧培养基37℃培养24h,分别加入刺五加苷B、刺五加苷E,培养后加甲醇提取离心,取上清液采用HPLC及UPLC/MS方法对代谢成分进行分离和定性分析。结果：刺五加苷B、苷E在大鼠粪便孵育液中代谢,24h后样本中能检测出较高浓度代谢物。在离体条件下,刺五加苷B、苷E可以被大鼠的肠道菌群代谢,经过UPLC/MS的检测,刺五加苷B代谢物的分子离子峰[M＋H]~＋为193.08,推测为刺五加苷B的苷元再脱一分子水;刺五加苷E代谢物的分子离子峰[M＋H]~＋为417.17,推测为刺五加苷B的苷元。结论：刺五加苷B、苷E可以被大鼠肠道菌群代谢。     

Comparison of in vitro preparations for semi-quantitative prediction of in vivo drug metabolism.

Various in vitro preparations were compared with respect to their ability to mimic in vivo metabolism. For this purpose, S9-liver homogenate, microsomes, cryopreserved hepatocytes, cryopreserved liver slices and fresh liver, lung, kidney, and intestinal slices were incubated with three drugs in development, which are metabolized in vivo by a wide range of biotransformation pathways. Metabolites were identified and quantified with liquid chromatography-mass spectometry/UV from the in vitro incubations and compared with metabolite patterns in feces, urine, and bile of dosed rats. In vitro systems with intact liver cells produced the same metabolites as the rat in vivo and are a valuable tool to study drug metabolism. Phase I metabolites were almost all conjugated in intact cells, whereas S9-homogenate only conjugated by sulfation and N-acetylation. Microsomes and S9-homogenate are useful to study phase I metabolism but not for the prediction of in vivo metabolism. Extra-hepatic organ slices did not form any metabolites that were not produced by liver cells, but the relative amounts of the various metabolites differed considerably. Small intestinal slices were more active than liver slices in the formation of the N-glucuronide of compound C, which is the major metabolite in vivo. When the relative contribution of liver and small intestinal slices to the metabolism of this compound was taken into account, it appeared that the in vivo metabolite pattern could be well predicted. Results indicate that for adequate prediction of in vivo metabolism, fresh or cryopreserved liver slices or hepatocytes in combination with slices of the small intestines should be used.     

Structural transformation of lignan compounds in rat gastrointestinal tract.

Structural transformation of arctiin and tracheloside, major components of seeds of Arctium lappa and Carthamus tinctorius, were investigated using rat gastric juice (pH 1.2-1.5) and rat large intestinal flora in vitro. Quantitative analysis of lignans and their metabolites was carried out by high performance liquid chromatography. Both lignans were stable in rat gastric juice and arctiin was rapidly transformed to arctigenin in rat large intestinal flora, followed by conversion to the major metabolite, 2-(3",4"-dihydroxybenzyl)-3-(3',4'-dimethoxybenzyl)-butyrolactone. On the other hand, tracheloside also decreased dependently with time and was converted to trachelogenin and its major metabolite, 2-(3",4"-dihydroxybenzyl)-3-(3',4'-dimethoxybenzyl)-2-hydroxybutyrola ctone. These experiments suggest that in the course of metabolism of lignans, firstly a cleavage of the glycosidic bond occurred and then demethylation of the phenolic methoxy group in the alimentary tract followed.     

大鼠和人肠道菌群对苯环喹溴铵代谢转化作用的研究

目的:研究大鼠和人肠道菌群对苯环喹溴铵(BCQB)代谢的影响。方法:采用体外肠道菌群代谢方法,将大鼠或人的粪便溶解于人工肠液中,搅拌混合后过滤,滤液加入1mg·mL-1的苯环喹溴铵溶液0.5mL,于37℃厌氧培养48h,采用高效液相色谱串联质谱法对大鼠或人肠内菌温孵液样品进行分离和定性分析,并设不加BCQB溶液的大鼠或人肠内菌温孵液为空白样品。结果:与空白样品比较,大鼠或人肠内菌温孵液样品中总离子流色谱中除BCQB峰外未见新增色谱峰,其质谱碎片信息显示只有BCQB的特征碎片离子,未检测到BCQB的代谢产物。结论:在离体条件下,BCQB不被大鼠和人的肠道菌群代谢。     

Disposition of flavonoids via enteric recycling: enzyme stability affects characterization of prunetin glucuronidation across species, organs, and UGT isoforms

We characterized the in vitro glucuronidation of prunetin, a prodrug of genistein that is a highly active cancer prevention agent. Metabolism studies were conducted using expressed human UGT isoforms and microsomes/S9 fractions prepared from intestine and liver of rodents and humans. The results indicated that human intestinal microsomes were more efficient than liver microsomes in glucuronidating prunetin, but rates of metabolism were dependent on time of incubation at 37 degrees C. Human liver and intestinal microsomes mainly produced metabolite 1 (prunetin-5- O-glucuronide) and metabolite 2 (prunetin-4'- O-glucuronide), respectively. Using 12 human UGT isoforms, we showed that UGT1A7, UGT1A8, and UGT1A9 were mainly responsible for the formation of metabolite 1, whereas UGT1A1, UGT1A8, and UGT1A10 were mainly responsible for the formation of metabolite 2. This isoform-specific metabolism was consistent with earlier results obtained using human liver and intestinal microsomes, as the former (liver) is UGT1A9-rich whereas the latter is UGT1A10-rich. Surprisingly, we found that the thermostability of the microsomes was isoform- and organ-dependent. For example, human liver microsomal UGT activities were much more heat-stable (37 degrees C) than intestinal microsomal UGT activities, consistent with the finding that human UGT1A9 is much more thermostable than human UGT1A10 and UGT1A8. The organ-specific thermostability profiles were also evident in rat microsomes and mouse S9 fractions, even though human intestinal glucuronidation of prunetin differs significantly from rodent intestinal glucuronidation. In conclusion, prunetin glucuronidation is species-, organ-, and UGT-isoform-dependent, all of which may be impacted by the thermostability of specific UGT isoforms involved in the metabolism.     

Identification of 14 quercetin phase II mono- and mixed conjugates and their formation by rat and human phase II in vitro model systems

In this study, the HPLC, UV-vis, LC-MS, and 1H NMR characteristics of 14 different phase II mono- and mixed conjugates of quercetin were determined, providing a useful tool in the identification of quercetin phase II metabolite patterns in various biological systems. Using these data, the phase II metabolism of quercetin by different rat and human liver and intestine in vitro models, including cell lines, S9 samples, and hepatocytes, was investigated. A comparison of quercetin phase II metabolism between rat and human liver and intestinal cell lines, S9, and hepatocytes showed considerable variation in the nature and ratios of quercetin conjugate formation. It could be established that the intestine contributes significantly to the phase II metabolism of quercetin, especially to its sulfation, that organ-dependent phase II metabolism in rat and man differ significantly, and that human interindividual variation is higher for quercetin sulfation than for glucuronidation or methylation. Furthermore, quercetin conjugation by different in vitro models from corresponding origins may differ significantly. The identification of the various mono- and mixed quercetin phase II conjugates revealed significant differences in phase II conjugation by a variety of in vitro models and led to the conclusion that none of the in vitro models converted quercetin to a phase II metabolite mixture similar to the in vivo plasma metabolite pattern of quercetin. Altogether, the identification of a wide range of phase II metabolites of quercetin as presented in this study allows the determination of quercetin phase II biotransformation patterns and opens the way for a better-funded assessment of the biological activity of quercetin in a variety of biological systems.     

肠道菌群调控骨代谢及中药应用研究进展

肠道菌群是定植在人体内复杂而庞大的微生物群落,肠道菌群及其代谢物短链脂肪酸在参与人体代谢、抵御外来致病菌以及调节免疫机制等方面发挥重要作用。近年来,不少研究发现肠道菌群与骨骼代谢密切相关。肠道菌群可通过营养吸收、短链脂肪酸生成、调节机体免疫、影响机体代谢等多种途径调控骨代谢,影响骨量变化。本文综述了肠道菌群影响骨代谢中骨量变化的潜在途径及作用机制,以及中药干预肠道菌群调控骨代谢的相关进展,以期为骨代谢相关疾病骨质疏松症的防治提供新思路。     

急进高原后肠道菌群介导的溴吡斯的明体内代谢研究

目的 探讨肠道菌群是否参与溴吡斯的明的代谢及急进高原后肠道菌群对其代谢产生的影响。方法 将大鼠随机分为平原给药组、平原空白组、高原给药组、高原空白组,共4组,每组6只大鼠。平原给药组大鼠禁食12 h后于上海（中国人民解放军海军军医大学）进行实验,分别将10.2 mg（约含溴吡斯的明2.3 mg）溴吡斯的明片剂粉末用水溶解后ig给药,于给药前（0 h）及给药后0.33、0.66、1.00、1.50、2.00、3.00、4.00、6.00、8.00、12.00、24.00 h由眼眶后静脉丛取血进行药动学分析。高原给药组大鼠于碌曲县海拔为4 300 m的高原地区,大鼠禁食12 h后开始药动学实验,过程同平原给药组。收集平原空白组和高原空白组大鼠的粪便,通过16S rRNA法分析急进高原后肠道菌群的变化情况;体外孵育实验结合LCMS/MS法分析肠道菌群与溴比斯的明的代谢关系。结果 急进高原后大鼠肠道菌群组成发生了变化,菌群种类和数量均减少;溴比斯的明与大鼠粪便体外孵育实验结果表明肠道微生物群参与溴吡斯的明吸收前的代谢,且在高原缺氧条件下,肠道微生物群对其代谢减慢;急进高原后溴吡斯的明药动学参数药时曲线下面积（AUC_0~t）增加了57.60%,清除率（CL）降低了35.94%。结论 肠道菌群参与溴吡斯的明吸收前代谢,可能与其生物利用度低有关;并且急进高原后肠道菌群介导的溴吡斯的明代谢减慢,从而增加其吸收。     

干预肠道菌群对肠易激综合征症状学的影响

目的探讨菌群失调与肠易激综合征（IBS）的关系及干预肠道菌群对肠易激综合征症状学的影响。方法@2010年12月至2011年12月我院确诊的腹泻型肠易激综合征（IBS．D）患者100例,其中85例完成试验,将85例完成试验者完全随机分为干预组（43例）和对照组（42例）。②入选后,每4—6周检测粪便菌群,出现菌群失调的患者,干预组给予菌群调节制剂治疗,对照组未给予治疗。③当出现症状时,记录患者IBS症状学积分,未出现症状要做无症状记录。结果①对照组发生菌群失调27例,其中出现腹泻症状者24例,症状发生率为88．9％（24／29）,干预组发生菌群失调29例,干预后出现腹泻症状者15例,症状发生率为51．7％（15／27）,对照组症状发生率明显高于干预组（P〈0．05）;②对照组发生症状时症状学总分、腹痛时间、腹痛频率及腹胀比例症状学积分明显高于干预组[分别为（10．4±3．3）分比（6．8±2．8）分、（2．0±1．0）分比（1．2±1．1）分、（2．0±1．0）分比（1．2±1．0）分、2分（1,3）比0分（0,1）],而大便形状异常、排便过程异常及黏液便比例差异无统计学意义（P〉0．05）。结论肠道菌群失调与IBS存在一定关系,肠道菌群失调可能是IBS重要的发病原因之一。因此,恢复正常肠道菌群有助IBS的预防和治疗。