Augmentative effect of tetrandrine on pentobarbital hypnosis mediated by 5-HT_(1A) and 5-HT_(2A/2C) receptors in mice

It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.

The 5-HT2c receptor gene Cys23Ser polymorphism influences the intravaginal ejaculation latency time in Dutch Caucasian men with lifelong premature ejaculation

It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2c receptor gene polymorphism and the duration of IELT in men with LPE. Therefore, a prospective study was conducted in 64 Dutch Caucasian men with LPE. Baseline IELT during coitus was assessed by stopwatch over a 1-month period. All men were genotyped for Cys23Ser 5-HT2c receptor gene polymorphism. Allele frequencies and genotypes of Cys and Ser variants of 5-HT2c receptor gene polymorphism were determined. Association between Cys/Cys and Ser/Ser genotypes and the natural logarithm of the IELT in men with LPE were.investigated. As a result, the geometric mean, median and natural mean IELT were 25.2, 27.0, 33.9s, respectively. Of all men, 20.0%, 10.8%, 23.1% and 41.5% ejaculated within 10, 10-20, 20-30 and 30-60s after vaginal penetration. Of the 64 men, the Cys/Cys and Ser/Ser genotype frequency for the Cys23Ser polymorphism of the 5-HT2c receptor gene was 81% and 19%, respectively. The geometric mean IELT of the wildtypes （Cys/Cys） is significantly lower （22.6s; 95% CI 18.3-27.8s） than in male homozygous mutants （Ser/Ser） （40.4s; 95% CI 20.3-80.4s） （P = 0.03）. It is concluded that Cys23Ser 5-HT2c receptor gene polymorphism is associated with the IELT in men with LPE. Men with Cys/Cys genotype have shorter IELTs than men with Ser/Ser genotypes.

The Apoptotic Effect of the Methanol Extract of <i>Polygonum cuspidatum</i>through Up-Regulation Death Receptor 5 and CHOP in HSC-2 Human Oral Cancer Cells

Polygonum cuspidatum is used as a traditional medicinal herb for the therapy of various diseases including several types of cancers. In the present study, we focused on addressing the anti-cancer activity and molecular mechanism of methanol extract of Polygonum cuspidatum (MEPC) in HSC-2 human oral cancer cells. The effect of MEPC on oral cancer cells was estimated by 3-(4,5-dimethylthiazol-20yl)-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium (MTS) assay, 4’-6-diamidino-2-phenylindole (DAPI) staining and Western blot analysis. MEPC inhibited the cell viability and induced apoptosis through the induction of death receptor (DR) 5. MEPC also increased the expression of C/EBP homologous protein/growth arrest and the DNA damage-inducible gene 153 (CHOP), a transcription factor induced by ER stress. Thus, we concluded that the induction of CHOP leading to DR5 up-regulation is required for the anti-cancer activity of MEPC in HSC-2 cells and MEPC may be a promising drug candidate for oral cancer.

New aspects of the C5a receptor

The process of apoptotic cell death for maintenance of cell homeostasis is now believed to be flexible. To examine the mechanism for this flexibility, the process of programmed cell death is sometimes divided into three phases: initiation, effector and execution. We have demonstrated that apoptotic cells commonly express a de novo synthesized C5a receptor (C5aR), which belongs to the G protein-coupled receptor (GPCR) family. A natural agnostic ligand of the C5aR, C5a, is produced from plasma C5 by C5 convertase in the early phase of acute inflammation. Although it is not realistic, we found that C5a can adjust apoptotic cell lifespan long. We recently have read interesting reports that apoptotic cells can release natural agnostic ligands at the initiation phase and corresponding GPCRs are already expressed on cell surfaces of apoptotic cells. Conversely, we found that apoptotic cells commonly release an alternative antagonistic/agnostic ligand of the de novo synthesized C5aR, ribosomal protein S19 (RP S19) polymer. The RP S19 polymer can adjust apoptotic cell lifespan short. Importantly, the C5a-dependent regulation is limited by the C5aR sensitization, but the RP S19 polymer-dependent regulation is unlimited by the C5aR desensitization. Therefore, we suggested that apoptotic cells commonly release agnostic ligands in the initiation phase that should lengthen intermittently a period of the initiation phase. Next, apoptotic cells commonly release antagonistic/agnostic ligands in the effector phase that should continue shortening a period of the effector phase. In addition, we know that an inherited erythroblastopenia is associated with mutations in the RP S19 gene. However, the roles of RP S19 in the formation of erythroblast-macrophage islands are not clearly understood. We recently have found that a different arm that the RP S19 polymer has connects the de novo synthesized C5aR on erythroblasts and the generally expressed C5aR on macrophages. Therefore, we suggested that apoptotic cells commonly release antagonistic/agnostic ligands in the execution phase that should continue connecting apoptotic cells and macrophages in the execution phase for shortening a period of the execution phase. In this review, we introduce new aspects of the C5aR in apoptotic cells and discuss the effects of the long lifespan of apoptotic cell-like neutrophils on the development of periodontitis.

Effects of Estradiol on 5-HTsA and 5-HT2c Receptor Immunolabeling in Rat Hippocampus

Steroid hormones participate in the modulation of serotonergic transmission, including the regulation of synthetic and metabolic enzyme production, as well as receptor and transporter activity. The changes of 5-HT5A and 5-HT2c immunolabeling induced by steroids in the hippocampus ofovariectomized rats were studied in this work. Densitometric analysis in rat hippocampi were carried out for adjacent brain coronal immunolabeled sections after treatment with subcutaneous injections of vehicle, estradiol, progesterone or the combination of both steroids in ovariectomized rats. Exposure to estradiol and the combination of estradiol and progesterone significantly reduced the 5-HT5A-like immunosignal in the CA 1 region while progesterone did not induce changes. On the other hand, exposure to the combination of estradiol and progesterone or estradiol alone increased the 5-HT2c immunosignal in the same region. These results indicate that estradiol is involved in the discrete regulation of serotonin receptors 5-HT5A and 5-HT2c in rat hippocampus.

Chimeric protein probes for C5a receptors through fusion of the anaphylatoxin C5a core region with a small-molecule antagonist

Blockade of the interaction of anaphylatoxin C5a with its receptor C5aR1 has been actively studied as a potential treatment for many inflammatory diseases;but current C5a antagonists exhibit inadequate potency and poor species cross-reactivity, and novel biochemical tools are needed to investigate whether the core region of C5a contains important interaction epitopes that can explain these limitations. Herein, we report the development of chimeric protein C5a probes containing both the complete core region of rat or human C5a, and the small-molecule antagonist PMX53-1. These probes were chemically synthesized through hydrazide-based native chemical ligation of a linear peptide hydrazide with the requisite cyclopeptidic antagonist, both of which were made by solid-phase synthesis. Quasi-racemic X-ray crystallography established that attachment of PMX53-1 did not affect the structure of the core region of C5a. Subsequent C5aR1 activity assays demonstrated the probes can provide valuable insights into the development of C5a antagonists;for example, they exhibited significantly better binding affinity and much improved species cross-reactivity than PMX53-1, supporting the notion that the effect of some epitopes outside the C-terminus of C5a should be taken into consideration when designing better C5a antagonists. Surprisingly, the core region of C5a was found to partially agonize C5aR1, suggesting the presence of more than one agonistic interaction in the binding of C5a to C5aR1. This study exemplifies the value of chemical protein synthesis in developing novel receptor probes for drug discovery research.

Bioassay-guided isolation of saikosaponins with agonistic activity on 5-hydroxytryptamine 2C receptor from Bupleurum chinense and their potential use for the treatment of obesity

5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was revealed to have agonistic activity on 5-HT2 C receptor, and the subsequent bioassay-guided isolation led to identification of several saikosaponins as the active constituents with 5-HT2 C receptor agonistic activity in vitro and anti-obesity activity in vivo. The new compound, 22-oxosaikosaponin d(1), was determined by extensive spectroscopic analyses(HR-ESI-MS, IR, and 1D and 2D NMR). The primary structure-activity relationship study suggested that the intramolecular ether bond between C-13 and C-28 and the number of sugars at C-3 position were closely related to the 5-HT2 C receptor agonistic activity. Saikosaponin a(3), the main saponin in B. chinense, showed obviously agonistic activity on 5-HT2 C receptor with an EC50 value of 21.08 ± 0.33 μmol×L^(–1) in vitro and could reduce food intake by 39.1% and 69.2%, and weight gain by 13.6% and 16.4%, respectively, at 3.0 and 6.0 mg×kg^(–1) in vivo. This investigation provided valuable information for the potential use of B. chinense as anti-obesity agent.

Stiffness-tuned and ROS-sensitive hydrogel incorporating complement C5a receptor antagonist modulates antibacterial activity of macrophages for periodontitis treatment

Periodontitis is admittedly a microbe-driven intractable infectious disease,in which Porphyromonas gingivalis(Pg)plays a keystone role.Pg can selectively impair the antimicrobial responses of periodontal resident macrophages including their phagocytic and bactericidal activity without interfering their proinflammatory activity,which leads to microflora disturbance,destructive periodontal inflammation and alveolar bone loss eventually.Here,an injectable ROS-sensitive hydrogel is developed for releasing active bone marrow-derived macrophages(named ex-situ macrophages hereafter)and a complement C5a receptor antagonist(C5A)to the gingival crevice.Through appropriately tuning the hydrogel stiffness,the phagocytic activity of these macrophages is greatly enhanced,reaching an optimal performance at the elastic modulus of 106 kPa.Meanwhile,C5A avoids undesired C5a receptor activation by Pg to ensure the bacterial killing activity of both the ex-situ and in-situ macrophages.Besides,the ROS-sensitive hydrogels show another distinct feature of decreasing the ROS level in periodontal niche,which contributes to the alleviated periodontal inflammation and attenuated bone loss as well.This study highlights the potential of utilizing hydrogels with tailored biomechanical properties to remodel the functions of therapeutic cells,which is expected to find wide applications even beyond periodontitis treatment.

Netrin-1 signaling pathway mechanisms in neurodegenerative diseases

Netrin-1 and its receptors play crucial roles in inducing axonal growth and neuronal migration during neuronal development.Their profound impacts then extend into adulthood to encompass the maintenance of neuronal survival and synaptic function.Increasing amounts of evidence highlight several key points:(1)Diminished Netrin-1 levels exacerbate pathological progression in animal models of Alzheimer’s disease and Parkinson’s disease,and potentially,similar alterations occur in humans.(2)Genetic mutations of Netrin-1 receptors increase an individuals’susceptibility to neurodegenerative disorders.(3)Therapeutic approaches targeting Netrin-1 and its receptors offer the benefits of enhancing memory and motor function.(4)Netrin-1 and its receptors show genetic and epigenetic alterations in a variety of cancers.These findings provide compelling evidence that Netrin-1 and its receptors are crucial targets in neurodegenerative diseases.Through a comprehensive review of Netrin-1 signaling pathways,our objective is to uncover potential therapeutic avenues for neurodegenerative disorders.

GPRC5A在喉癌发生中的生物学作用及机制研究

目的探讨G蛋白偶联受体C家族5A(Gprotein-coupled receptor family C,member 5,group A,GPRC5A)在喉癌组织中的表达特征,分析其在喉癌发展进程中的作用及机制。方法选取河北医科大学第二医院收治的32例经病理检查确定为喉鳞状细胞癌患者的喉癌组织及癌旁组织,采用免疫组织化学方法检测GPRC5A在喉癌组织及癌旁组织中的表达,并分析其表达特征与患者临床资料的相关性。采用定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)方法对GPRC5A在喉癌组织与癌旁组织的表达进行组织水平的验证,在人支气管上皮细胞系BEAS-2B、喉癌细胞系TU686及下咽癌细胞系Fadu中进行细胞水平的验证。构建GPRC5A过表达质粒p-GPRC5A,转染至喉癌TU686细胞中采用cell counting kit 8、Transwell方法检测GPRC5A基因过表达对喉癌TU686细胞增殖、侵袭及迁移的影响。Western blot检测GPRC5A基因过表达对细胞凋亡相关蛋白表达的影响。Western blot验证GPRC5A基因过表达对表皮生长因子受体(epidermal growth factor receptor,EGFR)/STAT3(epidermal growth factor receptor/signal transducer and activator of transcription 3)通路的影响。结果免疫组织化学结果表明,GPRC5A在喉癌组织中的阳性表达率显著低于癌旁组织(χ^(2)=14.190,P<0.001)。GPRC5A在不同肿瘤分期、淋巴结转移、分化程度喉癌组织中表达差异无统计学意义(P>0.05)。GPRC5A在喉癌组中的表达显著低于癌旁组(t=3.175,P=0.003)。GPRC5A在喉癌TU686和Fadu细胞中表达为BEAS-2B的(0.73±0.08)和(0.78±0.04)倍,差异有统计学意义(F=9.060,P=0.015);CCK8结果表明,48 h后p-GPRC5A组的OD值为0.76±0.03,显著低于NC组1.20±0.01和对照组1.30±0.08(F=69.970,P<0.001);Transwell实验表明,p-GPRC5A组细胞迁移的细胞数为138.70±10.97,显著低于对照组251.3±16.9和NC组247.7±17.5(F=5731.100,P<0.001)。p-GPRC5A组细胞侵袭的细胞数为113.00±10.21,显著低于对照组193.3±010.02和NC组190.00±7.90(F=8894.100,P<0.001)。Western blot结果表明,与对照组相比,Caspase3蛋白在p-GPRC5A组表达显著升高(F=78.880,P<0.001),Bcl-2蛋白在p-GPRC5A组的表达显著降低(F=125.820,P<0.001)。EGFR和p-STAT3蛋白在p-GPRC5A组的表达显著降低(F=27.573,P=0.001;F=60.614,P<0.001)。结论GPRC5A在喉癌组织及细胞系中低表达。GPRC5A基因过表达可抑制喉癌细胞增殖,迁移和侵袭,促进细胞凋亡发生,并抑制EGFR/STAT3通路的激活。

天然构象GPRC5D磷脂纳米盘的组装研究

将G蛋白偶联受体GPRC5D蛋白组装进磷脂纳米盘类膜体系内以维持蛋白天然构象的稳定性并对其进行生物活性的验证.首先通过蛋白表达和纯化得到了GPRC5D蛋白,然后以GPRC5D蛋白、膜支架蛋白(MSP)和磷脂按一定比例混合制备GPRC5D磷脂纳米盘.组装后GPRC5D蛋白依旧展现出与阳性抗体结合的能力,充分表明磷脂纳米盘能够有效保留GPRC5D蛋白的生物活性.该研究为制备GPRC5D蛋白磷脂纳米盘提供了新的思路和方法,为探究其蛋白的结构与功能和药物研发奠定基础.

ICU急性呼吸衰竭患者外周血HCAR、DcR3、AQP-5水平与机械通气撤机结局的关系及价值分析

目的 探究重症监护室(ICU)急性呼吸衰竭患者外周血超敏C反应蛋白与白蛋白比值(HCAR)、诱骗受体3(DcR3)、水通道蛋白-5(AQP-5)水平与机械通气撤机结局的关系及临床价值。方法 选取连云港市第二人民医院2018年8月—2021年8月ICU急性呼吸衰竭患者120例,均行机械通气治疗,在符合自主呼吸试验(SBT)指征且通过30 min SBT后撤机,根据撤机后48 h内是否再插管分为撤机成功组(87例)和撤机失败组(33例),撤机前采集外周血检测HCAR、DcR3、AQP-5水平,分析外周血HCAR、DcR3、AQP-5水平与撤机结局的关系及预测价值。结果 两组呼吸衰竭类型、合并器官功能障碍综合征(MODS)、肺部超声评分(LUS)、急性生理与慢性健康评价系统Ⅱ(APACHEⅡ)评分差异有统计学意义(P<0.05);撤机失败组外周血HCAR、DcR3高于撤机成功组,AQP-5低于撤机成功组(P<0.05);Pearson相关性分析显示外周血HCAR、DcR3与LUS、APACHEⅡ评分呈正相关,AQP-5与LUS、APACHEⅡ评分呈负相关(P<0.05);单因素、多因素分析均显示,外周血HCAR、DcR3、AQP-5影响撤机结局(P<0.05);外周血HCAR、DcR3、AQP-5预测撤机失败的截断值分别为4.10 mg/g、14.55μg/L、7.91μg/L,联合预测撤机失败的曲线下面积(AUC)为0.915(95%CI:0.850~0.958),大于各指标单独预测;以截断值为界分为低水平与高水平,外周血HCAR、DcR3高水平患者30 d生存率低于低水平患者,外周血AQP-5高水平患者30 d生存率高于低水平患者(P<0.05)。结论 ICU急性呼吸衰竭患者外周血HCAR、DcR3、AQP-5水平与撤机结局密切相关,联合检测可作为预测撤机失败的重要辅助手段,还能帮助临床判断死亡风险,为临床提供可靠的数据支持。

血清CCL5、P2X7R水平与大血管闭塞性急性缺血性脑卒中并发恶性脑水肿的关系

目的探讨血清C-C基序趋化因子配体5(CCL5)、嘌呤能2X7受体(P2X7R)水平与大血管闭塞性急性缺血性脑卒中(AIS-LVO)并发恶性脑水肿(MBE)的关系。方法选择AIS-LVO患者162例(观察组),接受静脉溶栓联合机械取栓治疗后72 h并发MBE 43例、未并发MBE 119例,同期另选体检健康的志愿者60例作为对照组。采集所有研究对象外周静脉血,离心留取血清,采用ELISA法检测血清CCL5、P2X7R。采用多因素Logistic回归模型分析AIS-LVO并发MBE的危险因素,采用受试者工作特征(ROC)曲线分析血清CCL5、P2X7R水平对AIS-LVO并发MBE的预测价值。结果观察组血清CCL5、P2X7R水平均高于对照组(P均<0.05)。单因素分析发现,AIS-LVO患者并发MBE者NIHSS评分、ASPECTS评分及血清CCL5、P2X7R水平均高于其未并发MBE者(P均<0.05);多因素Logistic回归分析发现,NIHSS评分增加及血清CCL5、P2X7R水平升高为AIS-LVO并发MBE的独立危险因素(P均<0.05)。ROC曲线分析发现,血清CCL5、P2X7R水平单独和联合预测AIS-LVO并发MBE的曲线下面积(AUC)分别为0.780、0.790、0.880,血清CCL5、P2X7R水平联合预测AIS-LVO并发MBE的AUC大于二者单独(P均<0.05)。结论血清CCL5、P2X7R水平升高是AIS-LVO并发MBE的独立危险因素;血清CCL5、P2X7R水平对AIS-LVO并发MBE均有一定预测价值,二者联合预测价值更高。

GPRC5A调控的ABCB1表达对肺腺癌增殖的影响

目的ATP结合盒B亚家族成员1(ATP binding cassette subfamily B member 1,ABCB1)的异常表达在多种癌症的发生发展中发挥关键作用。然而,G蛋白偶联受体C家族5组A型(G protein coupled receptor family C group5 type A,GPRC5A)调控的ABCB1表达对肺腺癌增殖的影响仍不清楚。本研究探讨了GPRC5A调控的ABCB1表达对肺腺癌增殖的影响。方法我们采用RT-PCR、Western-blot或免疫组化实验,分析ABCB1在肺腺癌细胞系、人肺腺癌组织以及GPRC5A基因敲除小鼠和野生型小鼠的气管上皮细胞和肺组织中的表达。采用细胞计数试剂盒-8(CCK-8)分析GPRC5A基因敲除小鼠气管上皮细胞对化疗药物的敏感性。采用皮下肿瘤形成实验探讨下调ABCB1表达是否可抑制体内肺腺癌增殖。采用免疫荧光和免疫沉淀实验研究GPRC5A和ABCB1之间潜在的调控关系。结果ABCB1在肺腺癌细胞系和人类肺腺癌组织中表达上调。GPRC5A基因敲除小鼠的气管上皮细胞及肺组织的ABCB1表达高于野生型小鼠。与GPRC5A野生型小鼠的气管上皮细胞相比,GPRC5A基因敲除小鼠的气管上皮细胞对塔立奇达和多柔比星更敏感。注射移植细胞28天后,接受ABCB1基因敲除细胞移植的GPRC5A-/-C57BL/6小鼠的肺肿瘤的体积和重量均明显低于野生型细胞移植小鼠(P=0.0043,P=0.0060)。此外,免疫荧光和免疫沉淀实验表明,GPRC5A通过直接结合方式调控ABCB1的表达。结论GPRC5A通过抑制ABCB1表达降低肺腺癌增殖。GPRC5A调节ABCB1表达的途径有待研究。

5-HT_2受体-磷脂酶C的激活易化大鼠基底外侧杏仁核突触可塑性的研究

为了探讨5-HT2受体激动剂盐酸2,5-二甲氧基-4-碘苯基丙烷(DOI)对杏仁核突触可塑性的调节作用,本研究在杏仁核脑片上记录基底外侧杏仁核(BLA)场电位,应用单串的θ频率波刺激(TBS)诱导突触可塑性,观察DOI对TBS诱导的突触可塑性的影响,及5-HT2受体拮抗剂、磷脂酶C抑制剂能否抑制DOI的作用。结果显示:单串的TBS刺激外囊,在BLA仅诱导约为10min的短时程增强。灌流液中加入100μmol/L DOI 20min,对基础的场电位没有作用。但在DOI存在的情况下,单串的TBS即可诱导长时程增强,强直刺激30min后,增强的场电位斜率仍维持在基础值的(162.5±9.7)%(n=9,P<0.01)。DOI对TBS诱导的突触可塑性的易化作用可被5-HT2A/2C受体拮抗剂ketanserin和PLC抑制剂U73122所抑制。以上结果提示5-HT2A/2C受体的激活可通过磷脂酶C通路易化杏仁核的突触可塑性。

大鼠脊髓后角内5型代谢型谷氨酸受体与C纤维的关系——免疫组织化学及电镜研究

目的 揭示大鼠脊髓后角内 5型代谢型谷氨酸受体 (mGluR5 )与一级传入C纤维的关系。 方法 在大鼠单侧足底注射福尔马林作为疼痛刺激 ,应用免疫组织化学技术光镜下观察疼痛对脊髓后角内mGluR5的影响 ;在大鼠蛛网膜下腔注射辣椒素 ,以毁损C纤维 ,用免疫电镜技术观察大鼠脊髓后角浅层内mGluR5阳性结构与C纤维间的超微结构关系。 结果 脊髓后角疼痛刺激侧Ⅰ～Ⅱ层内mGluR5免疫反应性较对照侧明显增强 ;在蛛网膜下腔注射辣椒素的大鼠 ,电镜下可见其后角浅层内大量变性的C纤维轴突终末与 1至多个含mGluR5的树突形成突触联系。 结论 提示脊髓后角浅层内mGluR5主要作为突触后受体参与介导和调节C纤维内对伤害性信息的传入。

CCR5第一、二胞外环特异性结合的拮抗短肽对TNBS诱导SD大鼠结肠炎的治疗作用

目的:研究C-C趋化因子受体5(CCR5)膜外第一、二胞外环(ECL1和ECL2)特异性结合的拮抗短肽对三硝基苯磺酸(TNBS)诱导的结肠炎模型大鼠的治疗作用与机制。方法:采用100 mg/kg TNBS诱导结肠炎SD大鼠模型;用不同剂量的2条拮抗短肽(ECL1:25、35和45 mg/kg;ECL2:15、25和35 mg/kg)分别作用于模型大鼠,观察其对大鼠疾病活动指数(DAI)、结肠大体损伤指数(CMDI)和组织病理学改变的影响,采用real-time PCR和Western blot法分别检测结肠组织TNF-α和COX-2的mRNA与蛋白表达水平。结果:与模型组相比,有效剂量的ECL2拮抗短肽HY治疗组大鼠疾病活动程度、肠道溃疡及病理组织学损伤均有明显减轻,各评分指数差异具有统计学意义(P<0.05);TNF-α和COX-2的蛋白和mRNA表达水平均明显下降(P<0.05)。ECL1拮抗短肽GH作用的大鼠结肠炎症状评分及TNF-α和COX-2炎症因子表达无明显改变。结论:ECL2拮抗短肽可能通过下调结肠黏膜TNF-α和COX-2的表达来缓解TNBS诱导的SD大鼠结肠炎,而ECL1拮抗短肽的作用不明显。

苯并吡喃[3,4-c]吡啶-5-酮类化合物的合成

以4-酮-3-甲酸甲酯哌啶盐酸盐为原料,通过分子间环加成和N-烷基化反应,合成了一系列潜在的多巴胺D4受体配基苯并吡喃[3,4-c]吡啶-5-酮类化合物,并用1HNMR,IR,ESI-MS,元素分析对其进行了表征.

3-(4-[^(18)F]氟苄基)-8-羟基-1,2,3,4-四氢苯并吡喃[3,4-c]吡啶-5-酮的放射化学合成

多巴胺D4受体是一种G蛋白偶联受体,在精神分裂症病因发展中起着重要作用。通过核医学显像仪器,利用PET显像剂可以确定受体在活体内的分布、含量变化等。本文用三氟甲基磺酸-4-三甲基铵苯甲醛作标记前体,采用“一锅法”制备了一种潜在的多巴胺D4受体PET显像剂3-(4-[18F]氟苄基)-8-羟基-1,2,3,4-四氢苯并吡喃[3,4-c]吡啶-5-酮(18F-FHTP)。其总的合成时间为105min,放射化学产率为19.7%,比活度大于120GBq/μmol。

转染结构性雄烷受体对丝裂霉素C和5-氮丙啶-3-羟甲基-1-甲基吲哚-4,7-二酮的细胞毒性的影响

研究丝裂霉素C(MMC)及其衍生物5-氮丙啶-3-羟甲基-1-甲基吲哚-4,7-二酮[5-(aziridin-1-yl)-3-hydroxymethyl-1-methylindole-4,7-dione,629]的细胞毒性,以及结构性雄烷受体(constitutive androstane receptor,CAR)转染对其生物学效应的影响。将质粒mCAR/pCR3转染HepG2细胞,经G418耐药性筛选获得转染CAR的g2car细胞,以转染空载体pCR3(HepG2/pCR3)作为对照。用RT-PCR检测质粒和CYP2B6 mRNA的表达,用MTT法评价MMC和629对g2car细胞和HepG2细胞在有氧和乏氧条件下的细胞毒性。RT-PCR检测到CAR和CYP2B6 mRNA在g2car细胞中有表达,在HepG2细胞中无表达;此外,在乏氧情况下,MMC和629的细胞毒性比在有氧情况下均有所增加(P<0.05),并且转染CAR以后,两者的细胞毒性均增加,但对MMC的影响较明显(P<0.05),对629的影响不明显(P>0.05)。提示CAR可在转录水平调节药物的代谢,提高药物的毒性;CYP2B6可以主要代谢MMC,但不主要代谢629。转染CAR基因可以增加细胞CYP2B6 mRNA的表达,并可引起MMC和629毒性的改变。