Inhibiting NF-κB increases cholesterol efflux from THP-1 derived-foam cells treated with AngⅡ via up-regulating the expression of ATP-binding cassette transporter A1

Objective： To study the role of nuclear factor-kappa B（NF- κB） in cholesterol efflux from THP-1 derived-foam cells treated with Angiotensin Ⅱ（Ang Ⅱ ）. Methods：Cultured THP-1 derived-foam cells were treated with Ang Ⅱ or preincubated with tosyl-phenylalanine chloromethyl-ketone（TPCK） NF- κB inhibitor. The levels of activated NF- κB in the cells were examined by sandwich ELISA, Cellular cholesterol content was studied by electron microscopy scanning and zymochemistry via fluorospectrophotometer and cholesterol effiux was detected by scintillation counting technique. ABCA1 mRNA and protein were quantified by RT-PCR and Western blotting. Results：Addition of TPCK to the cells before Ang Ⅱ stimulation attenuated the response of NF- κB p65 nuclear translocation induced by Ang Ⅱ and showed no peak in foam cells group and caused a reduction in cholesterol content and an increase in cholesterol efflux by 24.1%（P〈 0.05） and 41.1%（P〈 0.05） respectively, when compared with Ang Ⅱ group. In accordance, the ABCA1 mRNA and protein were increased by 30% and 19%（P 〈 0.05） respectively, when compared with Ang Ⅱ group. Conclusion：Ang Ⅱ can downregulate ABCA1 in THP-1 derived-foam cells via NF- K B, which leads to less cholesterol effiux and the increase of cholesterol content with the consequence of the promotion of atherosclerosis.

ATP-binding cassette subfamily B member 1 (ABCB1) and subfamily C member 10(ABCC1O) are not primary resistance factors for cabazitaxel

Introduction:ATP-binding cassette subfamily B member 1(ABCB1) and subfamily C member 10(ABCCIO) proteins are efflux transporters that couple the energy derived from ATP hydrolysis to the translocation of toxic substances and chemotherapeutic drugs out of cells.Cabazitaxel is a novel taxane that differs from paclitaxel by its lower affinity for ATP-binding cassette(ABC) transporters.Methods:We determined the effects of cabazitaxel,a novel tubulin-binding taxane,and paclitaxel on paclitaxelresistant,ABCB1-overexpressing KB-C2 and LLC-MDR1-WT cells and paclitaxel-resistant,ABCC10-overexpressing HEK293/ABCC10 cells by calculating the degree of drug resistance and measuring ATPase activity of the ABCB1 transporter.Results:Decreased resistance to cabazitaxel compared with paclitaxel was observed in KB-C2,LLC-MDR1-WT,and HEK293/ABCC10 cells.Moreover,cabazitaxel had low efficacy,whereas paclitaxel had high efficacy in stimulating the ATPase activity of ABCB1,indicating a direct interaction of both drugs with the transporter.Conclusion:ABCB1 and ABCC10 are not primary resistance factors for cabazitaxel compared with paclitaxel,suggesting that cabazitaxel may have a low affinity for these efflux transporters.

YTHDC1介导ABCB6上调诱导AD小鼠神经元细胞铁死亡促进认知功能障碍机制的实验研究

目的研究ATP结合盒B亚家族转运蛋白6(ATP-binding cassette subfamily B transporter 6,ABCB6)对阿尔兹海默症(Alzheimer’s disease,AD)小鼠认知功能障碍的影响及其可能的潜在调控分子机制。方法通过注射β-淀粉样蛋白(amyloidβ-protein,Aβ)构建体内AD小鼠模型;采用水迷宫测试和Y迷宫测试评估大鼠学习与记忆能力、空间探索能力。通过神经元HT22细胞和Aβ构建体外AD细胞模型;采用RNA免疫沉淀(RNA immunoprecipitation,RIP)分析YTH结构域包含蛋白1(YTH domain containing proteins 1,YTHDC1)与ABCB6的结合关系;实时定量聚合酶链反应(qRT-PCR)检测过表达和敲低转染效率;Western blot检测YTHDC1和ABCB6蛋白,以及铁死亡相关蛋白[溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)]表达水平;CCK-8检测细胞活力;检测丙二醛(malondialdehyde,MDA)、还原型谷胱甘肽(glutathione,GSH)、活性氧(reactive oxygen species,ROS)水平及Fe^(2+)含量。结果AD小鼠海马组织及Aβ诱导的HT22细胞中ABCB6 mRNA(3.51±0.17 vs 1.02±0.01,3.45±0.21 vs 1.02±0.01)和蛋白(3.25±0.14 vs 1.01±0.01,3.14±0.16vs 1.01±0.01)水平均显著上调,差异具有统计学意义(t=-46.238,-20.349;-50.468,-23.013,均P<0.001)。敲低ABCB6显著降低AD小鼠抵达平台的时间、到达平台距离,增加小鼠自发交替率和进入新异臂次数的比值(t=27.007,11.264,24.414,19.901,均P<0.001)。敲低ABCB6促进HT22细胞增殖,抑制Aβ诱导的MDA,Fe^(2+)水平上调和GSH水平下调,减少ROS生成,促进SLC7A11和GPX4蛋白表达(t=2.883~26.122,均P<0.05)。YTHDC1蛋白通过与ABCB6 mRNA结合促进其稳定性,上调ABCB6蛋白表达。敲低YTHDC1显著降低ABCB6蛋白水平(t=18.504,P<0.001),促进HT22细胞增殖,升高GSH含量及SLC7A11和GPX4蛋白水平,降低MDA和Fe^(2+)含量,抑制ROS生成(t=4.404~14.486,均P<0.05)。敲低YTHDC1可以改善AD小鼠学习与记忆能力和空间探索能力。过表达ABCB6可逆转敲低YTHDC1对HT22细胞铁死亡和AD小鼠认知功能障碍的影响。结论YTHDC1可能通过介导ABCB6上调,诱导神经元细胞铁死亡,进而促进AD小鼠的认知功能障碍发生。

LncRNA SNHG16 promotes colorectal cancer proliferation by regulating ABCB1 expression through sponging miR-214-3p

Mounting evidence indicates that long non-coding RNAs(lncRNAs)have critical roles in colorectal cancer(CRC)progression,providing many potential diagnostic biomarkers,prognostic biomarkers,and treatment targets.Here,we sought to investigate the role and underlying regulatory mechanism of lncRNA small nucleolar RNA host gene 16(SNHG16)in CRC.The expressions of SNHG16 in CRC were identified by RNA-sequencing and quantitative reverse transcription PCR.The functions of SNHG16 were explored by a series of in vitro and in vivo assays(colony formation assay,flow cytometry assay,and xenograft model).Bioinformatics analysis,RNA fluorescence in situ hybridization and luciferase reporter assay were used to investigate the regulatory mechanism of effects of SNHG16.SNHG16 was found to be significantly elevated in human CRC tissues and cell lines.Functional studies suggested that SNHG16 promoted CRC cell growth both in vitro and in vivo.Mechanistically,we identified that SNHG16 is expressed predominantly in the cytoplasm.SNHG16 could interact with miR-214-3p and up-regulated its target ABCB1.This study indicated that SNHG16 plays an oncogenic role in CRC,suggesting it could be a novel biomarker and therapeutic target in CRC.

The classification of ATP-binding cassette subfamily A member 3 mutations using the cystic fibrosis transmembrane conductance regulator classification system

Importance: The ATP-binding cassette subfamily A member 3 (ABCA3) protein plays a vital role in surfactant homeostasis. Mutations in the ABCA3 gene lead to the development of interstitial lung disease. In the most severe manifestation, mutations can lead to a fatal respiratory distress syndrome in neonates. ABCA3 belongs to the same ATP-binding cassette transporter superfamily as the cystic fibrosis transmembrane conductance regulator (CFTR), the gene that causes cystic fibrosis. Objective: To classify ABCA3 mutations in a manner similar to CFTR mutations in order to take advantage of recent advances in therapeutics. Methods: Sequence homology between the CFTR protein and the ABCA3 protein was established. The region of CFTR that is a target for the new potentiator class of drugs was of particular interest. We performed a literature search to obtain all published mutations that were thought to be disease causing. We classified these mutations using the established CFTR classification system. When possible, we drew on previous experimental classification of ABCA3 mutations. Results: Although the proteins share the same overall structure, only a 19％identity was established between CFTR and ABCA3. The CFTR therapeutic target region has a 22％ homology with the corresponding ABCA3 region. Totally 233 unique protein mutations were identified. All protein mutations were classified and mapped to a schematic diagram of the ABCA3 protein. Interpretation: This new classification system for ABCA3, based on CFTR classification, will likely aid further research of clinical outcomes and identification of mutation-tailored therapeutics, with the aim for improving clinical care for patients with ABCA3 mutations.

Efficacy of Shoushen granule on adenosine triphosphate binding cassette transporter A1, proprotein convertase subtilisin/kexin type 9 and toll-like receptor 4/nuclear factor kappa-B signaling pathway in ApoE-knockout mice

OBJECTIVE: To evaluate the efficacy of Shoushen granule, prepared with four Chinese medicinals, on the targeted regulation of adenosine triphosphate binding cassette transporter A1(ABCA1) through proprotein convertase subtilisin/kexin type 9(PCSK9) and toll-like receptor 4(TLR4)/nuclear factor kappa-B(NF-κB) signaling pathway to affect atherosclerosis(AS) in ApoE-knockout(ApoE-/-) mice.METHODS: ApoE-/-mice fed with a high-fat diet were used for AS modeling and divided into Model,Shoushen, and Atorvastatin groups. C57 BL/6 J mice at the same age and background strain were included in the Control group. Western blot and immunohistochemistry were used to measure ABCA1, PCSK9, TLR4, and NF-κB protein expression in mouse aortas. Enzyme-linked immuno sorbent assay was used to measure mouse serum tumor necrosis factor-α(TNF-α), interleukin-10(IL-10), monocyte chemoattractant protein 1(MCP-1), and intercellular cell adhesion molecule-1(ICAM-1) expression. Serum lipid profiles and histopathology were also assessed. Shoushen granule were composed of Heshouwu(Radix Polygoni Multiflori) 15 g, Gouqizi(Fructus Lycii) 15 g, Sheng shanzha(Raw Fructus Crataegus Pinnatifidae) 10 g, and Sanqi(Radix Notoginseng) 3 g.RESULTS: ApoE-/-mice fed with a high-fat diet had notable AS lesions, with reduced ABCA1 and IL-10 levels, elevated PCSK9, TLR4, NF-κB, TNF-α, MCP-1,and ICAM-1 expression, and increased total cholesterol(TC) and low density lipoprotein cholesterol(LDL-C) contents. With drug interventions, the areas of AS plaques were significantly reduced,the ABCA1 and IL-10 levels were increase, while the PCSK9, TLR4, NF-κB, TC, and LDL-C contents,and the TNF-α, MCP-1, and ICAM-1 expression were reduced.CONCLUSION: Shoushen granule effectively interfered with AS development by antagonizing the expression of key factors of the PCSK9 and TLR4/NF-κB signaling pathway to upregulate ABCA1 expression.

New insights into renal lipid dysmetabolism in diabetic kidney disease

Lipid dysmetabolism is one of the main features of diabetes mellitus and manifests by dyslipidemia as well as the ectopic accumulation of lipids in various tissues and organs,including the kidney.Research suggests that impaired cholesterol metabolism,increased lipid uptake or synthesis,increased fatty acid oxidation,lipid droplet accumulation and an imbalance in biologically active sphingolipids(such as ceramide,ceramide-1-phosphate and sphingosine-1-phosphate)contribute to the development of diabetic kidney disease(DKD).Currently,the literature suggests that both quality and quantity of lipids are associated with DKD and contribute to increased reactive oxygen species production,oxidative stress,inflammation,or cell death.Therefore,control of renal lipid dysmetabolism is a very important therapeutic goal,which needs to be archived.This article will review some of the recent advances leading to a better understanding of the mechanisms of dyslipidemia and the role of particular lipids and sphingolipids in DKD.

Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4^(-/-) mouse model

AIM: To study the interleukin-1(IL-1) pathway as a therapeutic target for liver fibrosis in vitro and in vivo using the ATP-binding cassette transporter b4^(-/-)(Abcb4^(-/-)) mouse model.METHODS: Female and male Abcb4^(-/-) mice from 6 to 13 mo of age were analysed for the degree of cholestasis(liver serum tests), extent of liver fibrosis(hydroxyproline content and Sirius red staining) and tissue-specific activation of signalling pathways such as the IL-1 pathway [quantitative polymerase chain reaction(q PCR)]. For in vivo experiments, murine hepatic stellate cells(HSCs) were isolated via pronasecollagenase perfusion followed by density gradient centrifugation using female mice. Murine HSCs were stimulated with up to 1 ng/m L IL-1β with or without 2.5 μg/m L Anakinra, an IL-1 receptor antagonist, respectively. The proliferation of murine HSCs was assessed via the Brd U assay. The toxicity of Anakinra was evaluated via the fluorescein diacetate hydrolysis(FDH) assay. In vivo 8-wk-old Abcb4^(-/-) mice with an already fully established hepatic phenotype were treated with Anakinra(1 mg/kg body-weight daily intraperitoneally) or vehicle and liver injury and liver fibrosis were evaluated via serum tests, q PCR, hydroxyproline content and Sirius red staining. RESULTS: Liver fibrosis was less pronounced in males than in female Abcb4^(-/-) animals as defined by a lower hydroxyproline content(274 ± 64 μg/g vs 436 ± 80 μg/g liver, respectively; n = 13-15; P < 0.001; MannWhitney U-test) and lower m RNA expression of the profibrogenic tissue inhibitor of metalloproteinase-1(TIMP)(1 ± 0.41 vs 0.66 ± 0.33 fold, respectively; n = 13-15; P < 0.05; Mann-Whitney U-test). Reduced liver fibrosis was associated with significantly lower levels of F4/80 m RNA expression(1 ± 0.28 vs 0.71 ± 0.41 fold, respectively; n = 12-15; P < 0.05; Mann-Whitney U-test) and significantly lower IL-1β m RNA expression levels(1 ± 0.38 vs 0.44 ± 0.26 fold, respectively; n = 13-15; P < 0.001; Mann-Whitney U-test). No gender differences in the serum liver parameters [bilirubin; alanine aminotransferase(ALT); aspartate aminotransferase and alkaline phosphatase(AP)] were found. In vitro, the administration of IL-1β resulted in a significant increase in HSC proliferation [0.94 ± 0.72 arbitrary units(A.U.) in untreated controls, 1.12 ± 0.80 A.U. at an IL-1β concentration of 0.1 ng/m L and 1.18 ± 0.73 A.U. at an IL-1β concentration of 1 ng/m L in samples from n = 6 donor animals; P < 0.001; analyses of variance(ANOVA)]. Proliferation was reduced significantly by the addition of 2.5 μg/m L Anakinra(0.81 ± 0.60 A.U. in untreated controls, 0.92 ± 0.68 A.U. at an IL-1β concentration of 0.1 ng/m L, and 0.91 ± 0.69 A.U. at an IL-1β concentration of 1 ng/m L; in samples from n = 6 donor animals; P < 0.001; ANOVA) suggesting an anti-proliferative effect of this clinically approved IL-1 receptor antagonist. The FDH assay showed this dose to be non-toxic in HSCs. In vivo, Anakinra had no effect on the hepatic hydroxyprolinecontent, liver serum tests(ALT and AP) and profibrotic(collagen 1α1, collagen 1α2, transforming growth factor-β, and TIMP-1) and anti-fibrotic [matrix metalloproteinase 2(MMP2), MMP9 and MMP13 ] gene expression after 4 wk of treatment. Furthermore, the hepatic IL-1β and F4/80 m RNA expression levels were unaffected by Anakinra treatment.CONCLUSION: IL-1β expression is associated with the degree of liver fibrosis in Abcb4^(-/-) mice and promotes HSC proliferation. IL-1 antagonism shows antifibrotic effects in vitro but not in Abcb4^(-/-) mice.

ABCB1基因C1236T多态性与小儿癫对药物反应的关系

目的探讨ABCB1基因C1236T单核苷酸多态性在中国儿童中的分布及其与抗癫药物反应性的关系。方法研究对象180例,包括健康对照组80例和癫组100例。根据患儿对抗癫药物的反应性将癫组分为耐药组(49例)和药物反应良好组(51例)。提取所有研究对象外周血基因组DNA,采用PCR扩增后继以限制性内切酶片段长度多态性(RFLP)鉴定ABCB1基因12号外显子C1236T多态性。测定该位点基因型频率和等位基因频率,并进行统计学分析。结果各组儿童ABCB1基因C1236T基因型频率的分布符合Hardy-Weinberg平衡,提示其来自同一孟德尔群体。健康对照组与药物反应良好组及耐药组,药物反应良好组与耐药组间基因型频率比较,差异均无统计学意义(Pa>0.05)。健康对照组与药物反应良好组及耐药组,药物反应良好组与耐药组间等位基因频率比较,差异亦无统计学意义(Pa>0.05)。结论本研究结果未能显示ABCB1基因C1236T位点多态性分布与小儿癫对药物治疗反应的表型之间存在相关性。

ABCB1基因C3435T多态性与癫痫耐药关系的Meta分析

目的运用Meta分析的方法综合评价ATP结合盒B亚家族成员1转运蛋白基因似BCBl）C3435T多态性与癫痫耐药的相关性。方法制定原始文献的纳入和排除标准及检索策略，检索中外文数据库，收集有关ABCB C3435T多态性与抗癫痫药物（AEDs）治疗反应相关性的研究报告。采用等位基因型fC（UST）以及共显性模型（CCVSTT，CTUSTT）、显性模型（CC＋CTVSTT）和隐性模型（ccVSCT＋TT）等基因遗传模型进行对比与定量综合分析，同时按人种所属地域（亚裔人与白种人、进行亚组分析。结果共23篇文献纳入Meta分析，包括3704例癫痫耐药患者和4160例治疗有效患者，入选文献无发表偏倚。统计分析显示：C3435T位点多态性与癫痫耐药无统计学关联：等位基因C与T相比，随机效应模型：OR=1．05，95％CI：0．94～1．18，P=-0．390。各基因型对比以及进行亚组分析后也未发现统计学关联。结论ABCBJC3435T多态性与癫痫耐药无关联。

抗结核药物性肝损伤危险因素及其与SLCO1B1/ABCB1基因多态性的关联性

目的分析不同抗结核治疗方案致药物性肝损伤(DILI)情况及其与肝脏药物转运体1B1(SLCO1B1)/多药耐药性蛋白1(ABCB1)基因多态性的关联性。方法回顾性分析2018年10月-2020年4月医院收治的621例接受抗结核治疗的肺结核患者临床资料,按是否发生DILI分为DILI组78例、非DILI组543例,测定两组抗结核治疗前后肝功能[丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、谷氨酰转肽酶(GGT)、碱性磷酸酶(ALP)、总胆红素(TBIL)]变化,分析两组的临床资料并筛选DILI的危险因素,以聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测SLCO1B1基因rs4149014位点及ABCB1基因rs2231142位点多态性,分析SLCO1B1/ABCB1基因多态性与DILI的相关性。结果78例出现DILI,发生率为12.56%(78/621),复治肺结核、耐多药肺结核患者DILI发生率高于初治肺结核患者,耐多药肺结核患者DILI发生率高于复治肺结核患者(P<0.05);78例DILI组抗结核治疗后ALT、AST、GGT、ALP、TBIL峰值均高于抗结核治疗前及非DILI组(P<0.01),非DILI组治疗前后ALT、AST、GGT、ALP、TBIL差异无统计学意义;年龄、肝病史、饮酒史、抗结核治疗时机为发生DILI的独立危险因素(P<0.05);DILI组SLCO1B1基因rs4149014位点的基因型TT、GT、等位基因T频率高于非DILI组,DILI组ABCB1基因rs2231142位点的基因型AC、等位基因A频率高于非DILI组(P<0.05);Logistic回归分析显示,SLCO1B1基因rs4149014位点的基因型GT、等位基因T及ABCB1基因rs2231142位点的基因型AC与肺结核患者DILI具有相关性(P<0.05)。结论抗结核药物治疗肺结核导致的DILI发生率高,SLCO1B1/ABCB1基因多态性也与DILI有密切关系,可能是DILI的易感基因。

P-糖蛋白表达阳性对癌痛病人舒芬太尼或喷他佐辛PCIA效果的影响

目的:评价P-糖蛋白表达阳性对癌痛病人舒芬太尼或喷他佐辛PCIA的效果。方法:本研究为回顾性队列研究。收集2020年1月至2024年1月于南京市溧水区人民医院进行治疗的癌痛病人的病历资料,年龄54~71岁,体质量49~67 kg,性别不限,TNM分期Ⅱ-Ⅳ期。采用免疫组织化学法测定肿瘤组织P-糖蛋白表达情况。将肿瘤组织P-糖蛋白表达阴性的病人分为2组:舒芬太尼组(S_(1)组)和喷他佐辛组(P_(1)组);将肿瘤组织P-糖蛋白表达阳性的病人分为2组:舒芬太尼组(S_(2)组)和喷他佐辛组(P_(2)组)。当VAS>5 cm时行PCIA,S_(1)组和S_(2)组采用枸橼酸舒芬太尼2μg/kg+托烷司琼10 mg;P_(1)组和P_(2)组采用喷他佐辛3 mg/kg+托烷司琼10 mg,4组均以生理盐水稀释成100 ml,负荷量5 ml,背景输注速率2 ml/h,PCA剂量1 ml,锁定时间10 min。镇痛期间VAS>3 cm时,静脉注射氟比洛芬酯50 mg补救。记录PCIA 4 h内、>4~12 h、>12~24 h及>24~48 h时段舒芬太尼、喷他佐辛用量和氟比洛芬酯使用情况。记录镇痛期间呼吸抑制(SpO_(2)<90%)、恶心呕吐、皮肤瘙痒及心动过缓等不良反应的发生情况。结果:最终纳入100例患者,每组25例。S_(1)组与S_(2)组各时段舒芬太尼用量、氟比洛芬酯使用率及镇痛期间呼吸抑制、恶心呕吐、皮肤瘙痒及心动过缓发生率比较差异均无统计学意义(P>0.05);与P_(1)组比较,P_(2)组PCIA 4 h内、>4~12 h及>24~48 h时段喷他佐辛用量升高(P<0.05),各时段氟比洛芬酯使用率和镇痛期间呼吸抑制、恶心呕吐、皮肤瘙痒及心动过缓发生率差异无统计学意义(P>0.05)。结论:P-糖蛋白表达阳性可能会减弱癌痛病人喷他佐辛PCIA的效果,而对舒芬太尼PCIA的效果可能无影响。

ABCB1基因多态性与难治性癫痫

近年来,难治性癫痫的病因与多药耐药基因以及多药耐药基因与抗癫痫治疗的因果关系越来越受到关注。P糖蛋白(P-glycopretein,P-gp)是由ATP结合盒B亚家族成员1转运蛋白基因(ATP-binding cassette subfamily B member 1 transporter gene,ABCB1)编码的产物。它不仅可以限制抗癫痫药物(antiepileptic drug,AED)的消化道吸收,而且可以在细胞和亚细胞水平调控药物在中枢神经系统的运输过程。除了生理和环境因素的影响,P-gp的功能和表达的变化可能主要取决于ABCB1基因的多态性,这是目前研究得最广泛、最深入的多药耐药机制。本文就目前ABCB1基因多态性与难治性癫痫的相关性研究进展作一综述。

ABCB1基因多态性与糖皮质激素性股骨头坏死的关联

目的分析三磷酸腺苷结合盒转运子B亚家族成员1(ABCB1)基因多态性与糖皮质激素性股骨头坏死(SANFH)的关联性。方法选取2017年01月-2019年01月厦门大学附属第一医院收治的长期使用糖皮质激素治疗的95例患者,其中发生股骨头坏死(ONFH)患者55例为坏死组,未发生ONFH患者40例为对照组,检测两组骨代谢指标[Ⅱ型胶原羧基端端肽(CTXⅡ)、抗酒石酸酸性磷酸酶5b(TRACP-5b)],血流流变学指标(血黏度高切、血黏度低切、血浆黏度及红细胞压积);采用Sanger测序法检测ABCB1基因C1236T、G2677T/A及C3435T位点单核苷酸多态性(SNPs)。结果坏死组CTXⅡ(209.51±10.37)ng/L、TRACP-5b(52.04±6.55)U/L水平及血黏度高切(5.08±0.77)、血黏度低切(11.38±2.74)、血浆黏度(2.23±0.30)mPa·s及红细胞压积(50.92±7.12)%均高于对照组(P<0.05);坏死组ABCB1基因C3435T位点CC基因型及C等位基因频率高于对照组(P<0.05)。结论糖皮质激素性股骨头坏死患者骨代谢和血流流变学指标水平升高,ABCB1 C3435T位点C等位基因与SANFH易感性有关。

Effect of Baizhu(Rhizoma Atractylodis Macrocephalae)extract on intestinal absorption of brucine and strychnine in vitro and in situ

OBJECTIVE:To investigate the antagonistic effect of the extract of Baizhu(Rhizoma Atractylodis Macrocephalae)(RAM)on the intestinal absorption of brucine and strychnine in Strychnos nux-vomica(NUX)and propose the mechanism of these effects.METHODS:The apparent permeability value(Papp)and absorption rate constant(Ka)were chosen as indices.The everted intestinal sac model and in situ single-pass intestinal perfusion model were used to study the effects of the RAM extract on the absorption of brucine and strychnine.To confirm the results,the brucine and strychnine concentrations in hepatic portal venous blood were determined.Western blotting was used to study P-glycoprotein(P-gp)expression in the Caco-2 cell line.RESULTS:Papp and Ka of brucine and strychnine were significantly increased in the presence of a P-gp inhibitor,but no significant increase was noted in the presence of a tight junction regulator.The RAM extract inhibited the absorption of brucine and strychnine and enhanced P-gp expression.CONCLUSION:The primary absorption mechanism for brucine and strychnine is passive transport,which is affected by P-gp.

Effect of decoction of Fuzheng Jiedu Xiaoji formula(扶正解毒消积方) plus chemoembolization on primary liver cancer in patients

OBJECTIVE:To investigate the effect of the decoction of Fuzheng Jiedu Xiaoji formula(扶正解毒消积方,FJXF)plus chemoembolization(TACE)on primary liver cancer(PLC)in patients,and study the underlying mechanism.METHODS:Patients with PLC who met the inclusion criteria were randomized into case group and control group.The case group was treated with FJXF combined with TACE.The control group was treated with TACE alone.The short-term clinical effect was evaluated;liver biochemistry,liver function index and multidrug resistance-associated indicators were detected.RESULTS:FJXF combined with TACE in the case group significantly increased the disease control rate than TACE alone in the control group(83.3%vs 61.1%).There was a reduction in the serum alpha-fetoprotein at 8 weeks after treatment in each group,while no difference between the two groups.The same trend can be observed for transaminase and direct bilirubin in both groups.In the case group,it showed a significant increase for albumin at 8 weeks after treatment,while no change in the control group.Multidrug resistanceassociated indicators for multidrug resistance protein 1 and p-glycoprotein were upregulated in the case group but remained stable in the control group.CONCLUSIONS:FJXF combined TACE had a better short-term effect than TACE alone in patients with PLC.The potential mechanism was probably associated with alleviated multidrug resistance induced by FJXF.Additionally,FJXF didn’t increase the risk of liver damage in the combined therapy.

谷胱甘肽过氧化物酶2在肺腺癌中的表达及临床意义

目的探讨谷胱甘肽过氧化物酶2(GPX2)在肺腺癌中的表达及临床意义。方法收集皖南医学院弋矶山医院病理科和徐州市中心医院病理科2010年1月至2013年12月存档的152例原发性肺腺癌及其癌旁正常组织标本蜡块。标本行免疫组织化学检测GPX2与ATP结合盒转运蛋白(ABCB6)在肺腺癌组织中的表达。正电子发射计算机断层成像(PET/CT)结果由患者术前检查获得。利用在线工具GEPIA、cBioPortal分析GPX2 mRNA在肺腺癌组织的表达。采用采取t检验比较GPX2表达量和18F-FDG的最大化标准摄取值在肺癌组织和正常组织的差异,采用Pearson相关性分析分析GPX2与ABCB6蛋白表达的相关性,采取χ2检验比较GPX2蛋白表达在不同临床病理特征之间的差异,同时应用Kaplan-Meier法分析肺腺癌患者的生存预后。结果基于TCGA和GTEx数据,GPX2 mRNA在肺腺癌组织(483例)的表达较正常组织(347例)明显升高,且与ABCB6表达水平呈正相关(r=0.57,P<0.001)。在本研究的样本中,89例(58.6%)肺腺癌组织的GPX2蛋白表达较正常组织升高(P<0.001),同样,GPX2与ABCB6蛋白表达水平正相关(r=0.697,P<0.001);其中,GPX2高表达患者(29例)的18F-FDG的最大化标准摄取值也高于GPX2低表达患者(26例),差异具有统计学意义(t=4.262,P<0.001)。虽然GPX2蛋白表达在不同临床病理特征之间的差异均无统计学意义(P>0.05),但是GPX2表达升高的患者较GPX2低表达患者无进展生存期减少[平均34.75个月(95%CI:29.746~39.747)vs 48.91个月(95%CI:41.033~56.779),P=0.009]。结论GPX2在肺腺癌发挥癌基因作用,其作用机制可能与ABCB6介导的糖酵解有关。

多个癫痫相关基因多态性与拉莫三嗪治疗儿童癫痫血药浓度的相关性

目的拉莫三嗪是儿童癫痫的一线治疗药物,其血药浓度与临床疗效个体差异较大,药效学和药动学通路多个相关基因突变可能是个体差异大的原因,本研究探讨代谢酶UGT1A4,转运体OCT1与ABCB1,受体SCN1A基因多态性与拉莫三嗪治疗儿童癫痫稳态血药浓度的相关性。方法于2018年1月—2019年6月纳入杭州市第一人民医院单用拉莫三嗪治疗的49例癫痫儿童,采集拉莫三嗪治疗达稳定剂量超过7 d以上患儿的稳态谷浓度血浆,使用高效液相色谱法测定拉莫三嗪血药浓度,直接测序法分析代谢酶UGT1A4(rs2011425)、转运体OCT1(rs628031)、ABCB1(rs1045642与rs1128503)和受体SCN1A(rs3812718)的基因分型。采用Kruskal-Wallis H检验或Mann-Whitney U检验探究不同基因分型与拉莫三嗪(LTG)血药浓度及标准化血药浓度相关性[即LTG血浆药物浓度除以每日每千克体重的服药剂量,CDR,(μg/mL)/(mg/kg)],P<0.05为具有显著相关性。结果转运体OCT1 rs628031与拉莫三嗪CDR具有显著相关性,其中AA+AG携带者CDR值显著高于GG携带者,分别是1.27(1.03,2.20)和0.69(0.57,1.02),P=0.002;但其他基因型与CDR值无显著相关性。结论OCT1 rs628031基因突变可能是拉莫三嗪血药浓度和临床疗效个体差异大的重要因素。