Progress in experimental models to investigate the in vivo and in vitro antidiabetic activity of drugs

Diabetes mellitus is one of the world's most prevalent and complex metabolic disorders,and it is a rapidly growing global public health issue.It is characterized by hyperglycemia,a condition involving a high blood glucose level brought on by deficiencies in insulin secretion,decreased activity of insulin,or both.Prolonged effects of diabetes include cardiovascular problems,retinopathy,neuropathy,nephropathy,and vascular alterations in both macro-and micro-blood vessels.In vivo and in vitro models have always been important for investigating and characterizing disease pathogenesis,identifying targets,and reviewing novel treatment options and medications.Fully understanding these models is crucial for the researchers so this review summarizes the different experimental in vivo and in vitro model options used to study diabetes and its consequences.The most popular in vivo studies involves the small animal models,such as rodent models,chemically induced diabetogens like streptozotocin and alloxan,and the possibility of deleting or overexpressing a specific gene by knockout and transgenic technologies on these animals.Other models include virally induced models,diet/nutrition induced diabetic animals,surgically induced models or pancreatectomy models,and non-obese models.Large animals or non-rodent models like porcine(pig),canine(dog),nonhuman primate,and Zebrafish models are also outlined.The in vitro models discussed are murine and human beta-cell lines and pancreatic islets,human stem cells,and organoid cultures.The other enzymatic in vitro tests to assess diabetes include assay of amylase inhibition and inhibition ofα-glucosidase activity.

Protective effects of Ecklonia cava extract on the toxicity and oxidative stress induced by hair dye in in-vitro and in-vivo models

Oxidative hair dyes containingρ-phenylenediamine(PPD)are reported to induce an allergic reaction by promoting oxidative stress when absorbed through the skin.Despite the associated risk,these hair dyes remain popular owing to their convenience and sharpness of color.This makes it important to minimize the cytotoxicity and oxidative stress induced by PPD-containing hair dyes.Ecklonia cava extract has been evaluated in different studies for its protective effects against external stress in fibroblasts and keratinocytes.Our study was aimed at using in-vitro and in-vivo models to investigate the extract’s effects on cytotoxicity of and oxidative stress induced by PPD-containing hair dyes.Analysis of CIEL*a*b*Color space was first used to determine the range of E.cava extract that would not interfere with the coloring ability of the dye upon addition.Subsequently,the set ranges of E.cava extract(5% and 7%)were added to the hair dye and their toxicity assessed by evaluating the viability of fibroblasts and keratinocytes.The effects on developmental phenotypes and induction of oxidative stress by hair dye were evaluated and compared with those of hair dyes containing different contents of E.cava extract using an in-vivo zebrafish model.Our results showed that E.cava extract in hair dye could significantly decrease the cytotoxicity and levels of oxidative stress caused by hair dyes containing PPD in both in-vitro and in-vivo models.These results suggest that the addition of 7% E.cava extract to 250μg/mL hair dye does not interfere with the coloring ability of the dye while showing significant protective eff ects against the hair dye.The study proposes that the use of E.cava extract as an adduct to hair dyes containing PPD reduces the cytotoxicity and oxidative stress induced by these hair dyes.

Design and comparative in-vitro and in-vivo evaluation of starch-acrylate graft copolymer based salbutamol sulphate sustained release tablets

The present work deals with the development of controlled release tablets of salbutamol sulphate(SS)using graft copolymers of methyl methacrylate(St-g-PMMA and Ast-g-PMMA)on starch and acetylated starch.Formulations were evaluated for physical characteristics like hardness,friability,drug release,drug content and weight variations,which fulfilled all the official requirements of tablet dosage form.The release rates from formulated matrix tablets were studied at SGF(pH 1.2)followed by SIF(pH 6.8).Drug release from the graft copolymer based tablets was found to be sustained upto the 14 h with>75%drug release.The in-vitro release study showed that the graft copolymer based matrix formulations(F3&F4)exhibited highest correlation value(r2)for higuchi kinetic model and Korsmeyer's model with n values between 0.61 and 0.67 proved that release mechanisms were governed by both diffusion and erosion mechanism.There was no significant difference in the pharmacokinetic parameters(tmax,Cmax,AUC,Ke,and t1/2)of the graft copolymers matrices and HPMC K100M matrix tablets,indicating their comparable sustained release effect.The potential of graft copolymers to sustain the drug release is well supported by in-vivo pharmacokinetic studies and their adequate physicochemical properties make them promising excipients for controlled drug delivery system.

In vitro-in vivo studies on anti-trypanosomal potentials of Zapoteca portoricensis

Objective:Aqueous extracts of Zapoteca portoricensis are used traditionally as antidiarrhea agent and in the treatment of diverse gastrointestinal disorders here in Nigeria specifically,the southern part.Similarly,the aqueous extract of the plant is also used traditionally as anticonvulsant,antispasmodic and in the treatment of tonsillitis.Recently too,the anti-inflammatory and antimicrobial activities of the methanol extracts of the root of Zapoteca portoricensis was reported.In this research,we are set to investigate the trypanocidal activity of Zapoteca portoricensis.Methods:The methanol extract of the root of Zapoteca portoricensis was investigated for both in vitro and in vivo trypanocidal activity following established models.In summary,phytochemical analysis was carried out on both the crude powdered root and on the methanol extract following standard procedures. The oral acute toxicity test(LD50 ) of the crude methanol extract was determined according to the method described by Lorke（1983）.Albino mice（17g-21g） of either sex were used.The methanol extract was suspended in 3%v/v tween 85 and administered orally at doses of 10 mg/kg,100 mg/kg and 1 000 mg/kg to three groups of mice（n = 3 ）.The animals were observed for 24 hours.Based on the result obtained in this initial test,doses of 4 mg/kg,6 mg/kg,and 8 mg/kg were administered to three different mice.The LD50 was calculated as the geometric mean of the lowest dose killing a mouse and the highest dose showing no death.The invivo /in-vitro antitrypanosomal evaluations were carried out in experimental animals and tissue cell culture respectively. Results:The result of the in vitro studies shows the inhibitive concentration-50（IC-50） against Trypanosoma brucei rhodesiense（T.b.rhodesiense） to be 0.372 mg/kg,while the control drug melarsoprol was 0.006 mg/kg.On Trypanosoma brucei brucei（T.cruzi）,the IC-50 is 6.42 mg/kg against 0.87 of the reference drug Benznidazole.The cytotoxicity on L-6 cells exhibited an IC-50 of 0.039 6 mg/kg against the reference drug,podophyllotoxin of 0.01 mg/kg.However,the in vivo study shows that the extract,at the administered doses,could not exhibit appreciable reduction of parasitemia and hence resulted to the death of test animals. Conclusion:The present data suggests that Zapoteca portoricensis could yield useful leads for the development of potentially potent antitrypanocides.

Various models of atrial fibrillation induced by acetylcholine and its application in the field of traditional Chinese medicine

房颤是一种严重危害社会公共健康的疾病,增加了发病率和死亡率,带来了巨大的经济负担.发病率包括增加了心血管结局事件比如心力衰竭和中风,严重影响了病人的生活质量、功能状态和认知.我们期待建立一种接近临床病理状态下的房颤模型来探索房颤的发病机制.乙酰胆碱作为迷走神经的递质,已建立起众多房颤模型,我们通过探讨目前已建立的乙酰胆碱房颤模型,寻求一种更接近生理状态的模型,并对其潜在机制进行讨论以观察药物对此机制的影响.

合成大麻素ADB-BUTINACA在人体尿液及体内外代谢模型中的代谢比较

本研究采用液相色谱-高分辨质谱法分析人体尿液样本、斑马鱼体内代谢模型以及肝微粒体体外代谢模型中N-(1-氨甲酰基-2,2-二甲基丙基)-1-丁基吲唑-3-甲酰胺(ADB-BUTINACA)的代谢情况。结果表明,共检测到45个ADB-BUTINACA代谢物,包括37个Ⅰ相代谢物和8个Ⅱ相代谢物,涉及9种代谢途径,其中有7个Ⅰ相代谢物为首次报道。通过对比代谢模型和人体尿液样本中的代谢物发现,斑马鱼体内代谢模型产生的代谢物更接近人体尿液样本,但无论是体内还是体外代谢模型,其代谢物排名与人体尿液样本具有较大差异,仅凭体内外代谢模型得到的代谢物数据难以直接用于真实样本的分析。通过对人体尿液样本中代谢物峰面积排名分析,建议将ADB-BUTINACA原型、M36、M19以及M16作为ADB-BUTINACA的生物标志物。

Hepatitis C virus: Morphogenesis, infection and therapy

Hepatitis C virus(HCV) is a major cause of liver diseases including liver cirrhosis and hepatocellular carcinoma. Approximately 3% of the world population is infected with HCV. Thus, HCV infection is considered a public healthy challenge. It is worth mentioning, that the HCV prevalence is dependent on the countries with infection rates around 20% in high endemic countries. The review summarizes recent data on HCV molecular biology, the physiopathology of infection(immune-mediated liver damage, liver fibrosis and lipid metabolism), virus diagnostic and treatment. In addition, currently available in vitro, ex vivo and animal models to study the virus life cycle, virus pathogenesis and therapy are described. Understanding of both host and viral factors may in the future lead to creation of new approaches in generation of an efficient therapeutic vaccine.

Experimental study on pharmacodynamics of pudilan xiaoyan oral liquid for preventing and treating respiratory virus infection

Background:Pudilan Xiaoyan Oral Liquid(PDL),a famous traditional Chinese formula for treating acute and chronic inflammation.To evaluate the broad-spectrum antiviral effect of Pudilan Xiaoyan Oral Liquid,and provide a basis for clinical medication.Methods:Its inhibitory effect on different respiratory viruses was observed by cytopathic test.The potential mechanism of the anti-influenza effect was determined by neuraminidase activity.In order to observe the therapeutic effect of PDL on viral pneumonia caused by different respiratory viruses.The viral pneumonia model was established by nasal infection with different respiratory viruses,and then PDL was given Therapeutic and prophylactically to evaluate its pharmacodynamic activity in vivo.Results:The results of in vitro experiments showed that PDL had different inhibitory effects on cytopathic effects caused by different respiratory viruses.And it has obvious inhibitory effect on the neuraminidase activity of influenza A virus,which indicates that it exerts anti-influenza virus effect by inhibiting neuraminidase activity of influenza virus.The results in vivo showed that PDL exhibited an inhibitory effect on pulmonary index(PI)and effectively reduced the degree of lesions in the lungs.The lethal rate of mice was significantly decreased while survival time of mice was dramatically increased by PDL treatment in comparison to infection control,respectively.Conclusions:Our study demonstrates that PDL had a significant protection and treatment effect for respiratory virus infection in vitro and in vivo.

SOCS1/JAK2/STAT3 axis regulates early brain injury induced by subarachnoid hemorrhage via inflammatory responses

The SOCS1/JAK2/STAT3 axis is strongly associated with tumor growth and progression,and participates in cytokine secretion in many diseases.However,the effects of the SOCS1/JAK2/STAT3 axis in experimental subarachnoid hemorrhage remain to be studied.A subarachnoid hemorrhage model was established in rats by infusing autologous blood into the optic chiasm pool.Some rats were first treated with JAK2/STAT3 small interfering RNA(Si-JAK2/Si-STAT3)or overexpression plasmids of JAK2/STAT3.In the brains of subarachnoid hemorrhage model rats,the expression levels of both JAK2 and STAT3 were upregulated and the expression of SOCS1 was downregulated,reaching a peak at 48 hours after injury.Simultaneously,the interactions between JAK2 and SOCS1 were reduced.In contrast,the interactions between JAK2 and STAT3 were markedly enhanced.Si-JAK2 and Si-STAT3 treatment alleviated cortical neuronal cell apoptosis and necrosis,destruction of the blood-brain barrier,brain edema,and cognitive functional impairment after subarachnoid hemorrhage.This was accompanied by decreased phosphorylation of JAK2 and STAT3 protein,decreased total levels of JAK2 and STAT3 protein,and increased SOCS1 protein expression.However,overexpression of JAK2 and STAT3 exerted opposite effects,aggravating subarachnoid hemorrhage-induced early brain injury.Si-JAK2 and Si-STAT3 inhibited M1-type microglial conversion and the release of pro-inflammatory factors(inducible nitric oxide synthase,interleukin-1β,and tumor necrosis factor-α)and increased the release of anti-inflammatory factors(arginase-1,interleukin-10,and interleukin-4).Furthermore,primary neurons stimulated with oxyhemoglobin were used to simulate subarachnoid hemorrhage in vitro,and the JAK2 inhibitor AG490 was used as an intervention.The in vitro results also suggested that neuronal protection is mediated by the inhibition of JAK2 and STAT3 expression.Together,our findings indicate that the SOCS1/JAK2/STAT3 axis contributes to early brain injury after subarachnoid hemorrhage both in vitro and in vivo by inducing inflammatory responses.This study was approved by the Animal Ethics Committee of Anhui Medical University and the First Affiliated Hospital of University of Science and Technology of China(approval No.LLSC-20180202)on March 1,2018.

Quantitative Mitochondrial Proteomics Study on Protective Mechanism of Grape Seed Proanthocyanidin Extracts Against Ischemia/Reperfusion Heart Injury in Rat

Cardiac ischemia/reperfusion（I/R） injury is a critical condition,often associated with high morbidity and mortality.The cardioprotective effect of grape seed proanthocyanidin extracts（GSPE） against oxidant injury during I/R has been described in previous studies.However,the underlying molecular mechanisms have not been fully elucidated.This study investigated the effect of GSPE on reperfusion arrhythmias especially ventricular tachycardia（VT） and ventricular fibrillation（VF）,the lactic acid accumulation and the ultrastructure of ischemic cardiomyocytes as well as the global changes of mitochondria proteins in in vivo rat heart model against I/R injury.GSPE significantly reduced the incidence of VF and VT,lessened the lactic acid accumulation and attenuated the ultrastructure damage.Twenty differential proteins related to cardiac protection were revealed by isobaric tag for relative and absolute quantitation（iTRAQ） profiling.These proteins were mainly involved in energy metabolism.Besides,monoamine oxidase A（MAOA） was also identified.The differential expression of several proteins was validated by Western blot.Our study offered important information on the mechanism of GSPE treatment in ischemic heart disease.

Modern approaches for detection of volatile organic compounds in metabolic studies focusing on pathogenic bacteria:Current state of the art

Pathogenic microorganisms produce numerous metabolites,including volatile organic compounds(VOCs).Monitoring these metabolites in biological matrices(e.g.,urine,blood,or breath)can reveal the presence of specific microorganisms,enabling the early diagnosis of infections and the timely implementation of tar-geted therapy.However,complex matrices only contain trace levels of VOCs,and their constituent com-ponents can hinder determination of these compounds.Therefore,modern analytical techniques enabling the non-invasive identification and precise quantification of microbial VOCs are needed.In this paper,we discuss bacterial VOC analysis under in vitro conditions,in animal models and disease diagnosis in humans,including techniques for offline and online analysis in clinical settings.We also consider the advantages and limitations of novel microextraction techniques used to prepare biological samples for VOC analysis,in addition to reviewing current clinical studies on bacterial volatilomes that address inter-species in-teractions,the kinetics of VOC metabolism,and species-and drug-resistance specificity.

In vitro and in vivo correlation for lipid-based formulations: Current status and future perspectives

Lipid-based formulations(LBFs)have demonstrated a great potential in enhancing the oral absorption of poorly water-soluble drugs.However,construction of in vitro and in vivo correlations(IVIVCs)for LBFs is quite challenging,owing to a complex in vivo processing of these formulations.In this paper,we start with a brief introduction on the gastrointestinal digestion of lipid/LBFs and its relation to enhanced oral drug absorption;based on the concept of IVIVCs,the current status of in vitro models to establish IVIVCs for LBFs is reviewed,while future perspectives in this field are discussed.In vitro tests,which facilitate the understanding and prediction of the in vivo performance of solid dosage forms,frequently fail to mimic the in vivo processing of LBFs,leading to inconsistent results.In vitro digestion models,which more closely simulate gastrointestinal physiology,are a more promising option.Despite some successes in IVIVC modeling,the accuracy and consistency of these models are yet to be validated,particularly for human data.A reliable IVIVC model can not only reduce the risk,time,and cost of formulation development but can also contribute to the formulation design and optimization,thus promoting the clinical translation of LBFs.

胆管癌实验模型研究进展

胆管癌是肝脏第二大常见的恶性肿瘤,早期症状不典型、恶性程度高、病死率高是其特点,目前全球范围内胆管癌的总体发病率逐年递增,且多数胆管癌患者的预后较差,迫切需要有效的诊断、治疗策略来改变这一现状。肿瘤建模是癌症研究的一大热点,建立胆管癌实验模型有助于深入了解、研究胆管癌的发生、发展、药物治疗效应等,对胆管癌的早期诊断及治疗具有重要意义。

阿尔茨海默病发病机制及体内外模型研究进展

阿尔茨海默病(AD)是一种以认知功能障碍为主要特征的神经退行性疾病,具体发病机制目前尚不清晰。随着我国人口老龄化越来越严重,患有AD人群的比例越来越高,亟需深入探究其发病机制并建立合适的疾病模型。本文从当前AD的发病假说和体内外模型进行了综述,详细介绍了神经细胞β-淀粉样前体蛋白(APP)转基因模型、D-半乳糖(D-gal)模型、D-半乳糖联合β-淀粉样蛋白(Aβ)模型等10种体内模型和3种体外模型(HT22细胞模型、PC12细胞模型和SH-SY5Y细胞模型),以期为AD疾病的基础研究和新药开发提供模型参考和研究思路。

Preclinical experimental models of drug metabolism and disposition in drug discovery and development

Drug discovery and development involve the utilization of in vitro and in vivo ex perimental models.Different models,ranging from test tube experiments to cell cultures,animals,healthy human subjects,and even small numbers of patients that are involved in clinical trials,are used at different stages of drug discovery and development for determination of efficacy and safety.The proper selection and applications of correct models,as well as appropriate data interpretation,are critically important in decision making and succesful advancement of drug candidates.In this review,we discuss strategies in the applications of both in vitro and in vivo.experimental models of drug metabolism and disposition.

关节腔注射用雷公藤甲素微球组织分布与代谢

目的考察关节腔注射用雷公藤甲素微球(TPL-MS)健康大鼠组织分布与体内外代谢行为。方法建立超高效液相色谱-串联质谱(UPLC-MS/MS)方法,测定不同时间点TPL-MS在大鼠心、肝、脾、肺、肾、血浆、关节的分布;以雷公藤甲素溶液(TPL-IN)为参比,通过测定TPL-MS在大鼠肝匀浆和肝微粒体酶中代谢后的剩余量,计算不同时间点雷公藤甲素代谢百分比;对健康大鼠进行关节腔给药,采用高分辨质谱分析TPL-MS在大鼠肝脏、胆汁、尿液和粪便中的代谢物种类和代谢速度。结果给药后30 min,TPL-IN在肝匀浆和肝微粒体中分别下降50.32%、49.47%,TPL-MS分别下降47.62%、46.36%;体内外共观察到4种代谢物,两种制剂体内代谢途径和代谢物种类相似,TPL-IN主要代谢时间集中在给药后8 h,TPL-MS可在给药后0~24 h持续检出代谢物;TPL-MS给药后第3天在关节组织内达到最高浓度,维持时长接近2周。结论2种制剂体外代谢规律基本一致,体内代谢途径和代谢物种类一致。与TPL-IN相比,TPL-MS的组织分布和代谢速度发生明显改变,关节滞留时间延长,体内代谢过程延缓。

抗痤疮药物模型研究进展

痤疮发病机制复杂,其药理研究模型主要分为体内模型和体外模型,且针对不同的发病机制采用的药理模型也不相同。本文针对目前痤疮研究现状,综述了药理模型及其应用进展,以利于在实际应用中选择合适的模型,为治疗痤疮药物和化妆品研发提供参考。

羟基红花黄色素A体外消化特性与体内代谢规律研究

目的研究羟基红花黄色素A(hydroxysafflor yellow A,HSYA)体外模拟胃肠道系统中的稳定性,口服后小鼠组织中的分布,以及HSYA跨小肠上皮细胞膜运输能力。方法体外模拟胃肠道消化实验采用紫外分光光度法检测体外模拟胃肠道消化分别为1和2h后的HSYA的全光谱特性以及高效液相色谱法(high performance liquid chromatography,HPLC)测定HSYA的含量,动物实验包括3种不同摄入方式:腹腔注射、灌胃及长期饲喂。腹腔注射和灌胃给药方式分别有两个时间点:1和2 h,剂量为0.01 mL/g;长期饲喂方式为含有2.5%红花小花饲喂10周,采集小鼠组织,样品处理采用6%高氯酸和甲醇沉淀蛋白法,利用HPLC测定样本中HSYA的含量。HSYA与小肠上皮细胞共培养不同时间后,吸取培养液,把细胞裂解后,通过HPLC测定培养液和裂解后的HSYA含量。结果体外消化实验结果表明,胃、肠道消化后的HSYA结构未被破坏,HSYA在胃肠道的稳定性较好;腹腔注射红花水提物后HSYA在小鼠血清、肝脏、肾、肺中的含量随着时间延长而下降,只有肌肉中HSYA的含量随着时间的延长而升高,而脾、睾丸、脑、心中均未检测到HSYA。灌胃和长期饲喂红花水提物后小鼠组织和血清中未检测到HSYA,表明HSYA无法经口服被吸收入血。通过细胞实验进一步证明了HSYA无法透过小肠上皮细胞而实现跨膜转运。结论本研究证明了HSYA在胃肠道中的稳定性较好,但口服后不能跨过肠道屏障被吸收入血;HSYA以注射方式入血后在组织中的分布存在一定差异。

基于体外发酵研究膳食纤维复合体对老年人肠道菌群的调节作用

该研究旨在探究膳食纤维复合体(Dietary Fiber Complex,DFC)对代表性人体肠道菌群及其代谢物的调节。采集10位65~75岁老年人的粪便,分性别开展DFC干预的体外发酵实验。结果发现,2%添加量的DFC组干预效果最好,其中女性组大肠杆菌属、拟杆菌属、双歧杆菌属、乳酸杆菌属的相对表达量为对照组的52.76%、43.40%、249.79%和232.99%,男性组大肠杆菌属、拟杆菌属、双歧杆菌属和乳酸杆菌属的相对表达量为对照组的44.02%、53.14%、228.71%和206.11%。显著差异的肠道菌群代谢物中,女性组的异丁酸、丙酸、甲酸相对丰度显著增加(P<0.01),蛋氨酸、组氨酸、β-葡萄糖、苯丙氨酸、苏氨酸、丙三醇、天冬氨酸、精氨酸、1-甲基组氨酸的相对丰度显著减少(P<0.01),男性组的异丁酸、丙酸、丁酸盐、甲酸相对丰度显著增加(P<0.01),组氨酸、β-葡萄糖、苯丙氨酸、苏氨酸和天冬氨酸相对丰度显著减少(P<0.01),二者潜在的代谢途径都为组氨酸代谢与苯丙氨酸、酪氨酸和色氨酸生物合成。研究结果表明DFC对人体肠道菌群及代谢物的调节作用明显,且在不同性别上潜在的代谢途径变化一致。

体外消化模型评估食物过敏原致敏性研究进展

食物过敏受到了人们的广泛关注,检测和评价食物过敏原的潜在致敏性日益受到重视。体外胃肠道消化实验是食物过敏原潜在致敏性评价体系的重要环节之一。该文系统介绍体外胃肠道消化模型的种类及其应用,并综述体外消化模型与体内消化模型相比的优缺点,最后对食物过敏原消化稳定性与潜在致敏性的关系进行探讨,以期为食物过敏原潜在致敏性评价研究中体外模拟消化方法的应用及消化结果分析提供参考。