促肾上腺皮质激素释放激素对人早期胎盘组织绒毛膜促性腺激素释放 …

为探讨人早期胎盘组织产生的促肾上腺皮质激素释放激素（ＣＲＨ）的生物学作用及其绒毛膜促性腺激素（ｈＣＧ）合成和释放的调控机制,在人早期胎盘组织体外孵育模型上研究了ＣＲＨ及其拮抗剂（α－ＨｅｌｉｃａｌＣＲＦ９－４１）对ｈＣＧ基础分泌和由ＧｎＲＨ刺激的ｈＣＧ分泌的影响。结果表明,１０＾－１０￣１０＾－７ｍｏｌ／Ｌ的外源性ＣＲＨ可明显抑制ｈＣＧ的基础释放,而以１０＾－７ｍｏｌ／Ｌ作用最显著;加入１０＾＝－     

亚硝酸盐对培养心肌细胞周期的影响

本文用流式细胞仪测定了ＮａＮＯ２对体外培养的Ｗｉｓｔａｒ乳鼠心肌细胞周期的影响。结果表明,１０＾－６ｍｏｌ／Ｌ ＮａＮＯ２引起Ｓ期细胞明显减少（Ｐ〈０．０５）;１０＾－８ｍｏｌ／Ｌ ＮａＮＯ２对其 显著（Ｐ〉０．０５）。应用「３Ｈ」ＴｄＲ掺入法测定了ＮａＮＯ２对心肌细胞增殖的作用。实验发现,１０＾－６ｍｏｌ／Ｌ ＮａＮＯ２明显抑制细胞增殖,而１０＾－９－１０＾７ｍｏｌ／Ｌ的ＮａＮＯ２则促进细胞增殖     

脱氢表雄酮对人早期胎盘孕酮分泌的影响

采用绒毛组织体外卵育模型,观察了脱氢表雄酮对人早期胎盘滋养层组织孕酮分泌的可能影响。结果表明,高浓度（１０＾－６ｍｏｌ／Ｌ以上）的脱氢表雄酮及其硫酸盐可以显著地抑制体外卵育的人早期胎盘滋养层组织的孕酮分泌（Ｐ〈０．００１）,低浓度则无效应。脱氢表雄酮及其硫酸盐抑制孕酮分泌的影响不依赖于其芳香化产物雌激素。芳香化酶抑制剂４－羟基雄烯二酮也能抑制孕酮分泌（Ｐ〈０．００１）,但其作用可被１７β－雌二醇逆     

组胺H2受体激动剂4—甲基组胺及其拮抗剂雷尼替丁对HL—60白…

在ＨＬ－６０白血病细胞的体外培养中,组胺Ｈ２受体激动剂４－甲基组胺（１０＾－８－１０＾－４ｍｏｌ／Ｌ）对ＨＬ－６０细胞的增殖具有轻微的抑制作用,而组胺Ｈ２受体拮抗剂雷尼替丁（１０＾－８－１０＾－４ｍｏｌ／Ｌ）对ＨＬ－６０细胞的增殖具有较强的抑制作用。但用１０＾－６ｍｏｌ／Ｌ的雷尼替丁预处理ＨＬ－６０细胞则可部分地拮抗４－甲基组胺（１０＾－８－１０＾４ｍｏｌ／Ｌ）对ＨＬ－６０白血病细胞的抑制作用。     

尾加压素增加血管平滑肌细胞粘着斑激酶的含量和活力

在体外培养的血管平滑肌细胞上,采用蛋白印迹杂交免疫沉淀的方法,研究尾加压素Ⅱ（ＵⅡ）与粘着斑激酶（ＦＡＫ）介导的信号传导之间的关系。结果表明,ＵⅡ在１０＾－８、１０＾－７和１０＾－６ｍｏｌ／Ｌ的深度时,可分别使ＦＡＫ活力增加２．２、３．０４和０．７３倍。加入ＵⅡ在（１０＾０７ｍｏｌ／Ｌ）５ｍｉｎ后,ＦＡＫ活力增加９５％,但与对照组相比无显著性差异（Ｐ〉０．０５０,刺激１０ｍｉｎ后,ＦＡＫ活力升高３     

17β-雌二醇诱导血管内皮细胞一氧化氮释放及其与细胞内钙的关系

利用小牛胸主动脉内皮细胞（ＢＡＥＣｓ）作为模型,观察１７β－雌二醇（Ｅ２）ＢＡＥＣｓ一氧化氮（ＮＯ）释放、一氧化氮合酶（ｅＮＯＳ）ｍＲＮＡ表达和细胞内钙（〔Ｃａ＾２＋〕ｉ）的影响,以及雌激素受体（ＥＲ）拮抗剂ｔａｍｏｘｉｆｅｎ和ＮＯＳ抑制剂（Ｌ－ＮＡＭＥ）的作用。结果显示,Ｅ２（１０＾－１２￣１０＾－８ｍｏｌ／Ｌ）呈尝试依赖性促进ＢＡＥＣｓ中ＮＯ的释放,以１０＾－８ｍｏｌ／Ｌ浓度Ｅ２处理ＢＡＥＣｓ     

乙酰胆碱对培养T细胞功能的作用

本文研究不同浓度（１０＾－１０－１０＾－４ｍｏｌ／Ｌ）乙酰胆碱（ＡＣｈ）对离体培养的大鼠脾脏Ｔ淋巴细胞增殖的影响,并探讨其作用机制。实验结果表明：１０＾－９－１０＾－４ｍｏｌ／Ｌ可显著增强Ｔ细胞由刀豆素Ａ（Ｃ－Ａ）诱导的增殖反应,以１０＾－７－１０＾－６ｍｏｌ／Ｌ时最强。淋巴细胞先用ＡＣｈ刺激１ｈ或者刺激１ｈ后洗弃ＡＣｈ,再用ＣｏｎＡ诱导６ｈ的Ｔ细胞的增殖。１０＾－７－１０－６ｍｏｌ／Ｌ阿托品可阻     

组胺H_2受体激动剂4－甲基组胺及其拮抗剂雷尼替丁对HL－60白血病细胞增殖的作用

在ＨＬ－６０白血病细胞的体外培养中,组胺Ｈ＿２受体激动剂４－甲基组胺（１０￣（－８）－１０￣（－４）ｍｏｌ／Ｌ）对ＨＬ－６０细胞的增殖具有轻微的抑制作用,而组胺Ｈ＿２受体拮抗剂雷尼替丁（１０￣（－８）－１０￣（－４）ｍｏｌ／Ｌ）对ＨＬ－６０细胞的增殖具有较强的抑制作用。但用１０￣（－６）ｍｏｌ／Ｌ的雷尼替丁预处理ＨＬ－６０细胞则可部分地拮抗４－甲基组胺（１０￣（－８）－１０￣（－４）ｍｏｌ／Ｌ）对ＨＬ－６０白血病细胞的抑制作用。     

内皮素对失血性休克鼠脑微循环的影响及尼莫地平,川芎…

利用大鼠颅骨开窗观察软脑膜微循环的方法研究了内皮素（ＥＴ－１）１０＾－１０－１０＾－７ｍｏｌ／Ｌ对软脑膜微循环的影响以及失血性体克时软脑膜对ＥＴ－１的反应性。并用１０＾－７ｍｏｌ／Ｌ造成失血性休克后脑血管痉挛的模型,观察尼莫地平、川芎嗪、６５４－２对内皮素引起血管痉挛的治疗作用。１０＾－９,１０＾－８和１０＾－７ｍｏｌ／Ｌ３种浓度ＥＴ－１可使脑膜小动脉、细动脉强烈收缩,收缩率分别为２７．７％、４６     

高血压大鼠红细胞膜Ca^2＋,Mg^2＋—ATP酶对Ca^2＋反应的变...

本实验观察了正常及高血压大鼠红细胞膜Ｃａ＾２＋,Ｍｇ＾２＋－ＡＴＰ酶对不同浓度Ｃｕ＾２＋Ｍｇ＾２＋的反应。结果表明：（１）在自发性高血压大鼠（ＳＨＲ）,此酶最适反应的Ｃａ＾２＋浓度为１０＾－６ｍｏｌ／Ｌ;ＷＫＹ大鼠为１０＾－４ｍｏｌ／Ｌ;两肾一环型肾性高血压大鼠（ＲＨＲ）为１０＾－７ｍｏｌｇ／Ｌ,Ｗｉｓｔａｒ大鼠为１０＾－４ｍｏｌ／Ｌ;Ｃａ＾２＋高于以上各相应浓度时该酶活性受到抑制;（２）作为该酶     

Phosphinic acid-based MMP-13 inhibitors that spare MMP-1 and MMP-3

Phosphinic acid-based inhibitors of MMP-13 have been investigated with the aim of identifying potent inhibitors with high selectivity versus MMP-1. Independent variation of the substituents on a P(1)' phenethyl group and a P(2) benzyl group improved potencies in both cases around 3-fold over the unsubstituted parent. Combining improved P(1)' and P(2) groups into a single molecule gave an inhibitor with a 4.5 nM IC(50) against MMP-13 and which is 270-fold selective over MMP-1.     

Expression of genes for metalloproteinases (MMP-1, MMP-2, MMP-9, and MMP-12) associated with psoriasis

Using real-time polymerase chain reaction (RT-PCR), we measured mRNA amounts of matrix metalloproteinases (MMPs): MMP-1, MMP-2, MMP-9, and MMP-12 genes in psoriatic lesions and unaffected skin of the same patients. We observed significant (about 15-fold) increase in the expression level of matrix metalloproteinase MMP-1 and MMP-12 genes associated with psoriasis. The results of our studies of MMP gene expression in cultured primary human keratinocytes treated with interleukin (IL-17) have shown upregulation of MMP gene expression both in cultured keratinocytes and in psoriatic skin lesions. Therefore, upregulation of MMP genes in the skin affected by psoriasis could result from IL-17 effects on skin cells.     

Orally bioavailable dual MMP-1/MMP-14 sparing,MMP-13 selective α-sulfone hydroxamates

A series of phenyl piperidine α-sulfone hydroxamate derivatives has been prepared utilizing a combination of solution-phase and resin-bound library technologies to afford compounds that are potent and highly selective for MMP-13, are dual-sparing of MMP-1 and MMP-14 (MT1-MMP) and exhibit oral bioavailability in rats.     

Isolation and characterization of human monoclonal antibodies specific to MMP-1A, MMP-2 and MMP-3

Matrix metalloproteinases (MMPs), a group of more than 20 zinc-containing endopeptidases, are up-regulated in many diseases, but the use of MMP inhibitors for therapeutic purposes has often been disappointing, possibly for the limited specificity of the drugs used in clinical trials. In principle, individual MMPs could be specifically drugged by monoclonal antibodies, either by inhibition of their catalytic activity or by antibody-based pharmacodelivery strategies. In this article we describe the isolation and affinity maturation of recombinant antibodies (SP1, SP2, SP3) specific to the murine catalytic domains of MMP-1A, MMP-2 and MMP-3. These novel reagents allowed a systematic comparative immunofluorescence analysis of the expression patterns of their cognate antigens in a variety of healthy, cancerous and arthritic murine tissues. While all three MMPs were strongly expressed in tumor and arthritis specimens, MMP-1A was completely undetectable in the normal tissues tested, while MMP-2 and MMP-3 exhibited a weak expression in certain normal tissues (e.g., liver). The new antibodies may serve as building blocks for the development of antibody-based therapy strategies in mouse models of pathology.     

MMP-1、MMP-3 和MMP-13在慢性睡眠剥夺所致颞下颌关节损伤中的变化

目的：观察MMP-1、MMP-3 和MMP-13 在慢性睡眠剥夺所致颞下颌关节损伤中表达的变化,探讨慢性睡眠剥夺所致颞下颌 关节损伤的可能机制。方法：采用改良多平台(MMPM)建立大鼠慢性睡眠剥夺模型,将90 只成年雄性Wastar 大鼠随机分为小平 台组、网格组和对照组。小平台组和网格组大鼠接受每天18 h的睡眠剥夺和6 h间歇期(10:00-16:00),间歇期大鼠正常笼养。实验 第7、14 和21 d时分别观察动物的行为学观察、检测动物血浆皮质醇(CORT)和促肾上腺皮质激素(ACTH)水平检测,通过免疫印 迹法和实时定量聚合酶链反应(PRC)检测颞下颌关节软骨中MMP-1、MMP-3 和MMP-13 的蛋白和mRNA表达,并通过HE 染色 法观察颞下颌关节结构的变化。结果：与对照组和网格组大鼠相比,小平台组大鼠第14 d和21 d 时髁突软骨中间部位表面纤维 在出现明显的炎症、松解及脱落现象;第21天时的血浆ACTH 和CORT 水平均显著高于网格组和对照组,差异有统计学意义 (P<0.05);第7、14、21 d时关节软骨MMP-1 和MMP-13 蛋白和mRNA 的表达水平均显著上调(P<0.05)。结论：慢性睡眠剥夺所致 的颞下颌关节损伤可能与关节软骨中MMP-1、MMP-3 和MMP-13 的表达上调有关。     

alpha-Amino-beta-sulphone hydroxamates as potent MMP-13 inhibitors that spare MMP-1

A series of alpha-amino-beta-sulphone hydroxamates was prepared and evaluated for potency versus MMP-13 and selectivity versus MMP-1. Various substituents were employed on the alpha-amino group (P(1) position), as well as different groups attached to the sulphone group extending into P(1)'. Low nanomolar potency was obtained for MMP-13 with selectivity versus MMP-1 of >1000x for a number of analogues.     

Association of MMP-2 and MMP-9 with clinical outcome of neurocysticercosis

Matrix metalloproteinases (MMPs) are the major endopeptidases involved in proteolysis of blood brain barrier (BBB) during central nervous system (CNS) infections. The present study detected serum levels and activities of MMP-2 and MMP-9 in patients with neurocysticercosis (NCC) and their association with symptomatic disease. In total, 68 individuals with NCC (36 symptomatic patients with active seizures and 32 asymptomatic individuals) and 37 healthy controls were enrolled for the study. Serum MMP-2 and MMP-9 levels and their activities were measured by ELISA and gel zymography respectively. Mean serum MMP-2 levels (ng/ml) were higher both in asymptomatic and symptomatic NCC cases compared to healthy controls. However, significantly higher levels of serum MMP-9 (ng/ml) were detected only in symptomatic NCC patients compared to asymptomatic NCC cases and healthy controls. Levels of both MMPs positively correlated with symptomatic NCC. Serum MMP-2 activities were significantly higher in symptomatic and asymptomatic NCC compared to healthy controls whereas serum MMP-9 activity was significantly associated with symptomatic NCC compared to healthy controls and asymptomatic NCC. In conclusion, the elevated level of MMP-9 in serum appears to play an important role in the development of symptoms i.e. active seizures in patients with NCC. However, further studies are needed to elucidate its precise role in disease pathogenesis.     

UVA-mediated downregulation of MMP-2 and MMP-9 in human epidermal keratinocytes

While human dermal fibroblasts increase the expression and secretion of distinct matrix metalloproteinases (MMPs) in response to ultraviolet (UV) irradiation, much less is known about regulation of MMPs with regard to normal human epidermal keratinocytes (NHEK). In this in vitro study, the effect of ultraviolet A (UVA) irradiation on gelatinase expression and secretion by NHEK was investigated. Irradiation of NHEK with non-toxic doses of UVA resulted in a dose-dependent downregulation of MMP-2 (gelatinase A) and MMP-9 (gelatinase B). A single dose of 30JUVA/cm(2) lowered MMP-2 activity to 26% and MMP-9 activity to 33% compared with mock-irradiated cells at 24h after irradiation. Downregulation of MMP-2 and MMP-9 steady-state mRNA levels was observed at 4h after UVA irradiation. The inhibitory effect of UVA on gelatinases was mediated by UVA-generated singlet oxygen (1O(2)). These findings suggest an inverse response to UVA irradiation in NHEK than in fibroblasts.     

WAVE3 promotes cell motility and invasion through the regulation of MMP-1, MMP-3, and MMP-9 expression

WAVE3 is a member of the WASP/WAVE family of proteins, which play a critical role in the regulation of actin polymerization, cytoskeleton organization, and cell motility. We show here that knockdown of the WAVE3 protein, using RNA interference in MDA-MB-231 cells, decreases phospho-p38 MAPK levels, but not those of phospho-AKT, phospho-ERK, or phospho-JNK. Knockdown of WAVE3 expression also inhibited the expression levels of MMP-1, MMP-3, and MMP-9, but not MMP-2. MMP production could be restored by PMA treatment, without affecting siRNA-mediated WAVE3 knockdown. The WAVE3-mediated downregulation of p38 activity and MMP production is independent of the presence of both WAVE1 and WAVE2, whose expression levels were not affected by loss of WAVE3. We also show that the downstream effect of the WAVE3 knockdown is the inhibition of cell motility and invasion, coupled with increased actin stress fiber formation, as well as reorganization of focal adhesion complexes. These findings suggest that WAVE3 regulates actin cytoskeleton, cell motility, and invasion through the p38 MAPK pathway and MMP production.     

Potent pyrimidinetrione-based inhibitors of MMP-13 with enhanced selectivity over MMP-14

Through the use of computational modeling, a series of pyrimidinetrione-based inhibitors of MMP-13 was designed based on a lead inhibitor identified through file screening. Incorporation of a biaryl ether moiety at the C-5 position of the pyrimidinetrione ring resulted in a dramatic enhancement of MMP-13 potency. Protein crystallography revealed that this moiety binds in the S(1)(') pocket of the enzyme. Optimization of the C-4 substituent of the terminal aromatic ring led to incorporation of selectivity versus MMP-14 (MT-1 MMP). Structure activity relationships of the biaryl ether substituent are presented as is pharmacokinetic data for a compound that meets our in vitro potency and selectivity goals.