Salvianolic acid B modulates the expression of drug-metabolizing enzymes in HepG2 cells

BACKGROUND: Enzymes involved in drug and xenobiotic metabolism have been considered to exist in two groups: phase I and phase II enzymes. Cytochrome P450 isoenzymes (CYPs) are the most important phase I enzymes in the metabolism of xenobiotics. The products of phase I metabolism are then acted upon by phase II enzymes, including glutathione S-transferases (GSTs). Herbs that inhibit CYPs such as CYP3A4 or that induce GSTs may have the potential to protect against chemical carcinogenesis since the mutagenic effects of carcinogens are often mediated through an excess of CYP-generated reactive intermediates. This study was designed to investigate the effects of salvianolic acid B (Sal B), a pure compound extracted from Radix Salviae Miltiorrhizae, a Chinese herb, on cell proliferation and CYP1A2 and CYP3A4 mRNA expression in the presence or absence of rifampicin, a potent inducer of CYPs and GST protein expression in HepG2 cells. METHODS: HepG2 cells were incubated with different concentrations of Sal B. Cell proliferation was determined by SYTOX-Green nucleic acid staining. CYP3A4 and CYP1A2 mRNA expression was assayed by real-time PCR. GST protein expression was analyzed by Western blotting. RESULTS: Low concentrations of Sal B (0-20 μmol/L) had no significant effects on cell proliferation, while higher concentrations (100-250 μmol/L) significantly inhibited proliferation in a concentration-dependent manner. Ten μmol/L Sal B, but not 1 μmol/L, down-regulated CYP3A4 and CYP1A2 mRNA expression after 24 hours of incubation, whereas both 1 and 10 μmol/L Sal B down-regulated CYP3A4mRNA expression after 96 hours of incubation; moreover, 1 and 10 μmol/L Sal B inhibited CYP3A4 mRNA expression induced by rifampicin. Both 1 μmol/L and 10 μmol/L Sal B increased GST expression. CONCLUSION: Sal B inhibits CYP3A4 and CYP1A2 mRNA expression and induces GST expression in HepG2 cells.

PXR/CAR-代谢酶/转运体解毒系统的功能及研究进展

目的综述孕烷X受体(PXR)和组成型雄甾烷受体(CAR)-代谢酶/转运体解毒系统的功能和研究进展。方法从肝脏代谢酶、肝肾转运体、PXR/CAR介导的药物解毒等多方面对PXR/CAR-代谢酶/转运体解毒系统及其在药物解毒方面的研究进展进行论述。结果与结论药物作为人体外源性物质,可能存在一定毒副作用,使用不当或毒性较强的药物易造成机体损伤。药物代谢酶和转运体是决定药物体内过程及暴露量的两大核心因素,在药物毒性与解毒方面扮演重要角色。随着对PXR和CAR研究的不断深入,发现其对药物解毒相关的代谢酶和转运体发挥核心调控作用,共同构成了机体最重要的防御解毒系统—PXR/CAR-代谢酶/转运体解毒系统。

Advances in Research of Mechanism of Herb-Drug Interactions

With the concurrent consumption of herbal medicines and conventional drugs,herb-drug interactions(HDIs)have become the most important clinical consequence of this practice.A general overview and the significance of pharmacokinetic and pharmacodynamic HDIs are provided,detailing basic mechanism,especially the metabolic enzymes and drug transporters,such as CYP450 and P-gp.

Role of farnesoid X receptor in establishment of ontogeny of phase-I drug metabolizing enzyme genes in mouse liver

The expression of phase-I drug metabolizing enzymes in liver changes dramatically during postnatal liver maturation.Farnesoid X receptor(FXR) is critical for bile acid and lipid homeostasis in liver.However,the role of FXR in regulating ontogeny of phase-I drug metabolizing genes is not clear.Hence,we applied RNA-sequencing to quantify the developmental expression of phase-I genes in both Fxr-null and control(C57BL/6) mouse livers during development.Liver samples of male C57BL/6 and Fxr-null mice at6 different ages from prenatal to adult were used.The Fxr-null showed an overall effect to diminish the "day-1 surge" of phase-I gene expression,including cytochrome P450 s at neonatal ages.Among the 185 phase-I genes from 12 different families,136 were expressed,and differential expression during development occurred in genes from all 12 phase-I families,including hydrolysis: carboxylesterase(Ces),paraoxonase(Pon),and epoxide hydrolase(Ephx); reduction: aldoketo reductase(Akr),quinone oxidoreductase(Nqo),and dihydropyrimidine dehydrogenase(Dpyd); and oxidation: alcohol dehydrogenase(Adh),aldehyde dehydrogenase(Aldh),flavin monooxygenases(Fmo),molybdenum hydroxylase(Aox and Xdh),cytochrome P450(P450),and cytochrome P450 oxidoreductase(Por).The data also suggested new phase-I genes potentially targeted by FXR.These results revealed an important role of FXR in regulation of ontogeny of phase-I genes.

Role of non-steroidal anti-inflammatory drugs on intestinal permeability and nonalcoholic fatty liver disease

The use of non-steroidal anti-inflammatory drugs(NSAIDs) is widespread worldwide thanks to their analgesic, anti-inflammatory and antipyretic effects. However, even more attention is placed upon the recurrence of digestive system complications in the course of their use. Recent data suggests that the complications of the lower gastro-intestinal tract may be as frequent and severe as those of the upper tract. NSAIDs enteropathy is due to enterohepatic recycling of the drugs resulting in a prolonged and repeated exposure of the intestinal mucosa to the compound and its metabolites. Thus leading to so-called topical effects, which, in turn, lead to an impairment of the intestinal barrier. This process determines bacterial translocation and toxic substances of intestinal origin in the portal circulation, leading to an endotoxaemia. This condition could determine a liver inflammatory response and might promote the development of nonalcoholic steatohepatitis, mostly in patients with risk factors such as obesity, metabolic syndrome and a high fat diet, which may induce a small intestinal bacterial overgrowth and dysbiosis. This alteration of gut microbiota may contribute to nonalcoholic fatty liver disease and its related disorders in two ways: firstly causing a malfunction of the tight junctions that play a critical role in the increase of intestinal permeability, and then secondly leading to the development of insulin resistance, body weight gain, lipogenesis, fibrogenesis and hepatic oxidative stress.

Effects of Buyang Huanwu Decoction on antioxidant and drug-metabolizing enzymes in rat liver

AIM: To study the effect of Buyang Huanwu Decoction(BYHWD) on the antioxidant enzymes and drug-metabolizing enzymes in rat liver. METHOD: Following treatment of rats with BYHWD at 6.42, 12.83, or 25.66 g·kg–1 per day for 15 days, microsomes and cytosols isolated from the liver tissues were prepared by differential centrifugation according to standard procedures. The activities of the antioxidant enzymes and cytochrome b5, NADPH-cytochrome P450 reductase, CYP3 A, CYP2E1, UGT, and GST of the rat livers were determined by UV-Vis spectrophotometer. RESULTS: The activities of ALT, AST, antioxidant enzymes, and the Hepatosomatic Index in serum were not significantly affected. In cytosols, the activity of CAT was significantly increased at the dosage of 12.83 g·kg–1, and all the other antioxidant activities and MDA levels were not affected by this treatment. BYHWD had no effect on cytochrome b5, NADPH-cytochrome P450 reductase, CYP3 A, and UGT. At the highest dose(25.66 g·kg–1), the activity of CYP2E1 was significantly inhibited, and the activities of GST and the level of GSH were increased. CONCLUSION: BYHWD is safe for the liver, and has the functions of detoxification and antioxidant. Patients should be cautioned about the herb-drug interaction of BYHWD and CYP2E1 substrates.

Characterizing drug-metabolizing enzymes and transporters that are bona fide CAR-target genes in mouse intestine

Intestine is responsible for the biotransformation of many orally-exposed chemicals.The constitutive androstane receptor(CAR/Nr1i3) is known to up-regulate many genes encoding drugmetabolizing enzymes and transporters(drug-processing genes/DPGs) in liver,but less is known regarding its effect in intestine.Sixty-day-old wild-type and Car / mice were administered the CARligand TCPOBOP or vehicle once daily for 4 days.In wild-type mice,Car m RNA was down-regulated by TCPOBOP in liver and duodenum.Car / mice had altered basal intestinal expression of many DPGs in a section-specific manner.Consistent with the liver data(Aleksunes and Klaassen,2012),TCPOBOP upregulated many DPGs(Cyp2b10,Cyp3a11,Aldh1a1,Aldh1a7,Gsta1,Gsta4,Gstm1-m4,Gstt1,Ugt1a1,Ugt2b34,Ugt2b36,and Mrp2–4) in specific sections of small intestine in a CAR-dependent manner.However,the m RNAs of Nqo1 and Papss2 were previously known to be up-regulated by TCPOBOP in liver but were not altered in intestine.Interestingly,many known CAR-target genes were highest expressed in colon where CAR is minimally expressed,suggesting that additional regulators are involved in regulating their expression.In conclusion,CAR regulates the basal expression of many DPGs in intestine,and although many hepatic CAR-targeted DPGs were bona fide CAR-targets in intestine,pharmacological activation of CAR in liver and intestine are not identical.

Impact of obese levels on the hepatic expression of nuclear receptors and drug-metabolizing enzymes in adult and offspring mice

The prevalence of obesity-associated conditions raises new challenges in clinical medication.Although altered expression of drug-metabolizing enzymes(DMEs)has been shown in obesity,the impacts of obese levels(overweight,obesity,and severe obesity)on the expression of DMEs have not been elucidated.Especially,limited information is available on whether parental obese levels affect ontogenic expression of DMEs in children.Here,a high-fat diet(HFD)and three feeding durations were used to mimic different obese levels in C57BL/6 mice.The hepatic expression of five nuclear receptors(NRs)and nine DMEs was examined.In general,a trend of induced expression of NRs and DMEs(except for Cyp2c29 and 3a11)was observed in HFD groups compared to low-fat diet(LFD)groups.Differentialeffects of HFD on the hepatic expression of DMEs were found in adult mice at different obese levels.Family-based dietary style of an HFD altered the ontogenic expression of DMEs in the offspring older than 15 days.Furthermore,obese levels of parental mice affected the hepatic expression of DMEs in offspring.Overall,the results indicate that obese levels affected expression of the DMEs in adult individuals and that of their children.Drug dosage might need to be optimized based on the obese levels.

Phenotype prediction of nonsynonymous single nucleotide polymorphisms in human phase II drug/xenobiotic metabolizing enzymes: perspectives on molecular evolution

Nonsynonymous single nucleotide polymorphisms (nsSNPs) in coding regions can lead to amino acid changes that might alter the protein’s function and account for susceptibility to disease and altered drug/xenobiotic response. Many nsSNPs have been found in genes encoding human phase II metabolizing enzymes; however, there is little known about the relationship between the genotype and phenotype of nsSNPs in these enzymes. We have identified 923 validated nsSNPs in 104 human phase II enzyme genes from the Ensembl genome database and the NCBI SNP database. Using PolyPhen, Panther, and SNAP algorithms, 44%?59% of nsSNPs in phase II enzyme genes were predicted to have functional impacts on protein function. Predictions largely agree with the available experimental annotations. 68% of deleterious nsSNPs were correctly predicted as damaging. This study also identified many amino acids that are likely to be functionally critical, but have not yet been studied experimentally. There was significant concordance between the predicted results of Panther and PolyPhen, and between SNAP non-neutral predictions and PolyPhen scores. Evolutionarily non-neutral (destabilizing) amino acid substitutions are thought to be the pathogenetic basis for the alteration of phase II enzyme activity and to be associated with disease susceptibility and drug/xenobiotic toxicity. Furthermore, the molecular evolutionary patterns of phase II enzymes were characterized with regards to the predicted deleterious nsSNPs.

m^(6)A修饰对药物代谢酶和药物转运体的调控作用

m^(6)A修饰是RNA甲基化修饰中最丰富的一种修饰,受甲基转移酶和去甲基化酶的动态调控,被m^(6)A识别蛋白识别并结合后可影响mRNA的剪切、稳定性和翻译等生物学过程来调控靶基因的表达。最近的研究发现,m^(6)A修饰可通过多种途径来调控药物代谢酶和药物转运体表达,进而影响机体对药物的代谢速率或影响细胞中的药物浓度,最终导致药物治疗效果发生变化。该文综述了m^(6)A修饰调控药物代谢酶和药物转运体分子机制的研究进展,以期为临床上的合理用药、个体化用药提供新思路。

高血压药物基因多态性分析对安徽皖南地区高血压患者个体化治疗的参考价值

目的:分析皖南地区高血压患者β受体阻滞剂、血管紧张素受体拮抗剂(ARB)、血管紧张素转换酶抑制剂(ACEI)、钙离子拮抗剂及利尿剂药物基因多态性分布频率,为高血压药物基因检测工作和个性化用药提供理论依据。方法:搜集2021年7月至2023年4月皖南医学院弋矶山医院839例高血压住院患者药物基因检测信息,分析各基因位点分布频率。结果:ACE(I/D)I/I、I/D及D/D基因型频率分别为42.1%、46.0%、11.9%;肾上腺素能受体β1(ADRB1)(1165G>C)G/G、G/C及C/C基因型频率分别为8.3%、40.0%、51.6%;血管紧张素Ⅱ受体1(AGTR1)(1166A>C)A/A、A/C及C/C基因型频率分别为90.2%、9.8%、0.0%;CYP2C9*3(1075A>C)*1/*1、*1/*3及*3/*3基因型频率分别为91.3%、8.7%、0.0%;CYP2D6*10(100C>T)*1/*1、*1/*10及*10/*10基因型频率分别为25.0%、36.6%、38.4%;CYP3A5*3(6986A>G)*1/*1、*1/*3及*3/*3基因型频率分别为7.0%、39.0%、54.0%;钠尿肽前体蛋白A(NPPA)(2238T>C)T/T、T/C及C/C基因型频率分别为97.9%、2.1%、0.0%。此外,皖南地区多个药物相关基因位点基因型分布频率与中国其他地区存在统计学差异(P<0.05)。结论:皖南地区高血压药物相关基因位点基因型分布频率具有一定偏向性,且与中国其他地区相比存在统计学差异,提示开展高血压药物基因多态性检测对皖南地区高血压药物个体化应用具有一定指导意义。

肝病患者药物代谢酶细胞色素P450酶1A2、多态性N-乙酰化酶、黄嘌呤氧化酶活性的变化

目的 :以咖啡因为代谢探针 ,研究肝病患者咖啡因代谢物比值 (caffeinemetaboliteratios ,CMRs)的变化 ,探讨CMRs作为药物代谢酶细胞色素P45 0酶 1A2 (CYP1A2 )、多态性N 乙酰基转移酶 (NAT2 )、黄嘌呤氧化酶 (XO)的活性指标 ,用于调整某些药物用药方案的可行性。方法 :选择健康受试者 30例 ,慢性肝病受试者 30例。采用高效液相色谱 (HPLC)法测定受试者尿液中 5种咖啡因主要代谢物浓度 ,计算特定的CMRs。对研究结果进行统计学处理 ,对健康受试组与肝病受试组、慢性活动性肝炎受试组、代偿期肝硬化受试组的CMRs数据分别进行t检验 ,分别考察组间数据的差异性 ,判断肝病患者药物代谢酶CYP1A2、NAT2、XO的酶活性变化。结果 :肝病患者包括慢性活动性肝炎或代偿期肝硬化患者CYP1A2、NAT2活性显著降低(P <0 .0 5 ) ;而XO活性无显著性变化 (P >0 .0 5 )。结论 :可为临床调整经去甲基化。

LC-MS/MS法研究Cocktail探针药物咖啡因、右美沙芬、奥美拉唑、咪达唑仑及其代谢产物在大鼠血浆中的药代动力学

目的建立灵敏、快速、稳定的测定大鼠血浆中CYP450探针药物/代谢产物,咖啡因/副黄嘌呤、奥美拉唑/5-羟基奥美拉唑、右美沙芬/右啡烷、咪达唑仑/1'-羟基咪达唑仑的LC-MS/MS方法,并用于其在大鼠体内的药代动力学研究。方法 100μL血浆样品加入内标地西泮及乙酸铵溶液,乙酸乙酯进行液液萃取,离心取上清挥干复溶,LC-MS/MS检测。选用Agilent Eclipse Plus-C_(18)色谱柱(50 mm×2.1mm,3.5μm),乙腈和0.01%甲酸(1 mmol·L^(-1)甲酸铵)作为流动相,梯度洗脱,流速为0.3 m L·min^(-1),进样量10μL。采用电喷雾电离源(ESI),扫描方式为多反应正离子监测模式(MRM+),咖啡因m/z:195.0/138.1;副黄嘌呤m/z:181.1/124.1;奥美拉唑m/z:346.1/198.1;5-羟基奥美拉唑m/z:362.1/214.1;右美沙芬m/z:272.2/147.1;右啡烷m/z:258.1/157.1;咪达唑仑m/z:326.1/291.1;1'-羟基咪达唑仑m/z:342.1/324.1;内标地西泮m/z:285.1/154.0。结果咖啡因、副黄嘌呤、奥美拉唑、5-羟基奥美拉唑、右美沙芬、右啡烷、咪达唑仑和1'-羟基咪达唑仑的线性范围分别为1.95~2 000、0.98~250、0.48~2 000、0.98~250、0.98~2 000、0.48~125、1.95~2 000和1.95~250μg·L^(-1)。RSD<15%且无基质效应。结论该方法快速、灵敏、重现性好,可用于血浆中上述探针药物/代谢产物浓度的测定,便于深入进行代谢酶CYP1A2、CYP2C19、CYP2D6和CYP3A4的代谢相关研究。

非甾体抗炎药疗效影响因素的研究进展

非甾体抗炎药(NSAIDs)是治疗骨关节炎、类风湿关节炎和急慢性疼痛最常用的药物之一。在使用NSAIDs的患者中发现,NSAIDs的疗效在个体之间存在较大差异,这可能导致患者治疗失败或出现危及生命的药物不良反应。本综述从药物代谢酶CYP2C9基因多态性、环氧化酶基因多态性以及肠道微生物群差异的角度对引起NSAIDs疗效差异的原因进行分析讨论,为NSAIDs个体化用药方案的制定提供参考依据。

基于液相色谱-质谱联用的肝微粒体中药物代谢酶定量方法研究进展

肝微粒体中的药物代谢酶是参与药物代谢的主要酶类,不管是合成药物,还是组成复杂的中药,主要经过肝脏中药物代谢酶的代谢转化成易于排泄的化合物。由于药物代谢酶的表达水平具有较大的个体差异性,因此,药物代谢酶的准确定量对药物的药理、药物相互作用和临床应用具有重要意义。本文综述了近十年来药物代谢酶定量方法的研究进展。

高原低氧影响心血管药物体内代谢的研究进展

高原低氧环境显著影响机体心血管系统,使体内药物代谢受到一定影响。药物的药动学参数、药物代谢酶活性和药物转运体的活性及表达均会发生改变。该文系统回顾高原低氧对心血管系统药物代谢的研究进展,探讨高原低氧条件下药动学、药物代谢酶和药物转运体的变化,以期为高原低氧环境下药物代谢的研究提供参考。

基于非天然氨基酸的蛋白质合成研究进展

蛋白质是生命体的重要组成部分,几乎参与了所有的生命活动。通常情况下生物体中蛋白质由20种天然氨基酸组成,但随着现代科技的飞速发展,人们对医疗、食品、材料等方面的需求日渐增大,这些蛋白质已经无法满足多样化的应用需求。随着非天然氨基酸及其合成的蛋白质的优异性能,使得利用非天然氨基酸改造蛋白质的技术逐渐成为研究热点。非天然氨基酸有着不同于天然氨基酸的侧链基团,可以给蛋白质带来新的性质与功能。该文基于对非天然氨基酸及非天然蛋白质的研究,首先介绍了以化学及生物的手段合成非天然氨基酸的方法,同时对化学及生物法的优劣势及未来的发展前景进行了总结;然后介绍了将非天然氨基酸掺入蛋白质的几种热门方法;接着对非天然氨基酸及其合成蛋白质的应用在酶的改造、科研以及医疗方面进行了列举和描述;最后对这项技术未来的发展进行了展望与总结。

降压治疗对原发性高血压患者血小板α颗粒膜蛋白-140及血管性假血友病因子(vWF)浓度的影响

目的:探讨降压治疗对原发性高血压(EH)患者血浆血小板α颗粒膜蛋白-140(GMP-140)及血管性假血友病因子(vWF)浓度的影响。方法:用放射免疫和酶联免疫吸附双抗夹心法分析法对30例EH患者分别于降压药物治疗前后测定外周静脉血浆GMP140及vWF浓度,并与30例健康体检者(对照组)相比较。结果:EH患者降压治疗前红细胞比容(Hct%)与血浆GMP140及vWF浓度比正常对照组明显升高;转化酶抑制剂(ACEI)与钙通道阻断剂(CCB)联合降压治疗6周后与治疗前相比明显降低;收缩压、舒张压与红细胞计数,红细胞含量,血小板计数无显著相关性,而与Hct%,GMP140及vWF呈显著正相关。结论:EH患者体内存在PTS与血小板活化和内皮功能损伤有关,ACEI与CCB联合降压治疗能改善血栓前状态。

3'-大豆苷元磺酸钠对肝药酶活性的影响

目的:探讨3'-大豆苷元磺酸钠对肝药酶活性的影响。方法:(1)腹腔注射3'-大豆苷元磺酸钠(0.1 mg/kg、0.2 mg/kg),15 min后,腹腔注射阈上剂量的戊巴比妥钠(2.5 g/L,0.02 mL/g),观察小鼠翻正反射消失时间;(2)腹腔注射3'-大豆苷元磺酸钠(0.1 mg/kg、0.2 mg/kg),15 min后,腹腔注射阈下剂量的戊巴比妥钠(2.5 g/L,0.01 mL/g),观察小鼠翻正反射消失时间;(3)连续腹腔注射3'-大豆苷元磺酸钠(0.1 mg/kg、0.2 mg/kg),连续10 d。取肝脏观察小鼠肝脏系数和肝药酶(肝微粒体细胞色素P450的含量变化)含量的影响。结果:(1)3'-大豆苷元磺酸钠能显著加速阈上剂量戊巴比妥钠的入睡时间和延长戊巴比妥钠的睡眠时间。(2)3'-大豆苷元磺酸钠能显著加速阈下剂量戊巴比妥钠的入睡时间和延长戊巴比妥钠的睡眠时间。(3)3'-大豆苷元磺酸钠能提高小鼠肝药酶含量。结论:3'-大豆苷元磺酸钠对肝药酶具有诱导作用。

经CYP 450酶代谢的常见阿片类药物比较

目的总结常见阿片类药物羟考酮、美沙酮、曲马多和丁丙诺啡涉及细胞色素P450(CYP450)酶的代谢过程。方法采用计算机检索PubMed自建库起至2024年3月关于羟考酮、美沙酮、曲马多和丁丙诺啡等阿片类药物经CYP450酶的代谢研究,以及CYP450酶涉及的药物相互作用。结果CYP450酶是羟考酮、美沙酮、曲马多和丁丙诺啡阿片类药物代谢过程中最重要的酶。CYP3A4和CYP2D6不仅参与大多数阿片类药物代谢,还参与大量其他药物的代谢,阿片类药物在用药过程中易与其他药物发生相互作用。结论了解阿片类药物在肝脏中的代谢,以及药物间相互作用涉及重叠的药效动力学、药代动力学,可避免用药过程中可能发生的药品不良反应。