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Evidence for the presence of peroxisome proliferator-activated receptor (PPAR) α and γ and retinoid Z receptor in cartilage -: PPARγ activation modulates the effects of interleukin-1β on rat chondrocytes

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第16期275卷 12243-12250页

作者： Bordji, K Grillasca, JP Gouze, JN Magdalou, J Schohn, H Keller, JM Bianchi, A Dauça, M Netter, P Terlain, B Univ Nancy 1 Fac Med Pharmacol Lab CNRSUHPUMR 7561 F-54505 Vandoeuvre Les Nancy France Univ Nancy 1 Fac Sci Lab Biol Cellulaire Dev Unite Propre Rech Enseignement Super 2402 F-54500 Vandoeuvre Les Nancy France

Peroxisome proliferator-activated receptor (PPAR) alpha, PPAR gamma, and retinoid acid receptor-related orphan receptor (ROR) alpha are members of the nuclear receptor superfamily of ligand-activated transcription factors. Although they play a key role in adipocyte differentiation, lipid metabolism, or glucose homeostasis regulation, recent studies suggested that they might be involved in the inflammation control and especially in the modulation of the cytokine production. This strongly suggests that these transcriptional factors could modulate the deleterious effects of interleukin-1 (IL-1) on cartilage. However, to date, their presence in cartilage has never been investigated. By quantitative reverse transcription-polymerase chain reaction, Western blot, and immunocytochemistry analysis, we demonstrated, for the first time, the presence of PPAR alpha, PPAR gamma, and ROR alpha in rat cartilage, at both mRNA and protein levels. Comparatively, the PPAR alpha mRNA content in cartilage was much lower than in the liver but not significantly different to that of the adipose tissue. PPAR gamma mRNA expression in cartilage was weak, when compared with adipose tissue, but similar to that found in the liver. ROR alpha mRNA levels were similar in the three tissues. mRNA expression of the three nuclear receptors was very differently modulated by IL-1 or mono-iodoacetate treatments. This indicates that they should be unequally involved in the effects of IL-1 on chondrocyte, which is in accordance with results obtained in other cell types. Indeed, we showed that 15d-PGJ2 mainly, but also the drug troglitazone, that are ligands of PPAR gamma could significantly counteract the decrease in proteoglycan synthesis and NO production induced by IL-1. By contrast, PPAR alpha ligands such as Wy-14,643 or clofibrate had no effect on this process. Therefore, the presence of PPAR gamma in chondrocytes opens up new perspectives to modulate the effects of cytokines on cartilage by the use of

关键词：藻酸盐软骨关节/化学软骨关节/代谢软骨细胞/化学软骨细胞/代谢氯贝丁酯/代谢内皮生长因子/代谢葡糖醛酸己糖醛酸类白细胞介素1/代谢配体褪黑激素/代谢核受体亚家族1 F组成员1 聚合酶链反应嘧啶类/代谢 RNA 信使/代谢大鼠 Wistar 受体细胞表面/分析受体细胞表面/代谢受体胞质和核/分析受体胞质和核/代谢受体褪黑激素受体维甲酸反式激活因子类转录因子/分析转录因子/代谢动物男(雄)性大鼠

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Peroxisomal targeting signal-1 receptor protein PEX5 from Leishmania donovani -: Molecular, biochemical, and immunocytochemical characterization

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第18期275卷 13637-13644页

作者： Jardim, A Liu, W Zheleznova, E Ullman, B Oregon Hlth & Sci Univ Dept Biochem & Mol Biol Portland OR 97201 USA

The human pathogens of the Leishmania and Trypanosoma genera compartmentalize glycolytic and other key metabolic pathways in unique subcellular microbodies called glycosomes, organelles related to the peroxisomes of mammals and yeast. The molecular machinery that carries out the specific targeting of glycosomal proteins to the organelle has not been characterized, although the bulk of glycosomal proteins contain the COOH-terminal tripeptide glycosomal peroxisomal targeting signal-1 (PTS-1) similar to the mammalian and fungal peroxisomal targeting signal. To characterize the mechanisms of glycosomal targeting, the gene encoding PEX5, designated LdPEX5, has been isolated from Leishmania donovani. LdPEX5 encodes a 625-amino acid protein with a molecular mass of 69.7 kDa. Like its homologs in yeast and humans, LdPEX5 predicts a protein with seven copies of a tetratricopeptide repeat in its COOH-terminal half proposed to mediate PTS-1 binding and three copies of a WXXX(Y/F) motif in its NH2 terminus conjectured to be essential for protein translocation into the organelle. LdPEX5 was overexpressed in Escherichia coli and purified to homogeneity for binding experiments and generation of antibodies. Recombinant LdPEX5 bound xanthine phosphoribosyltransferase (XPRT), a PTS-1 containing glycosomal protein with a K-D of 4.2 nM, but did not bind an XPRT in which the PTS-1 had been deleted. Moreover, binding studies with the COOH-terminal half of the LdPEX5 confirmed that this portion of the PEX5 protein was capable of binding the XPRT PTS-1 with an affinity of 17.3 nM. Confocal microsocopy revealed that LdPEX5 was predominantly in the cytosolic milieu, and genetic analysis implied that LdPEX5 was an essential gene.

关键词：氨基酸序列克隆分子杜氏利什曼原虫/代谢分子序列数据受体胞质和核/分析受体胞质和核/遗传学受体胞质和核/代谢序列比对序列分析动物人类

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The generation and characterization of a cell line derived from a sporadic renal angiomyolipoma - Use of telomerase to obtain stable populations of cells from benign neoplasms

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AMERICAN JOURNAL OF PATHOLOGY 2001年第2期159卷 483-491页

作者： Arbiser, JL Yeung, R Weiss, SW Arbiser, ZK Amin, MB Cohen, C Frank, D Mahajan, S Herron, GS Yang, JW Onda, H Zhang, HB Bai, XH Uhlmann, E Loehr, A Northrup, H Au, P Davis, I Fisher, DE Gutmann, DH Emory Univ Sch Med Dept Dermatol Atlanta GA 30322 USA Emory Univ Sch Med Dept Pathol Atlanta GA 30322 USA Winship Canc Inst Atlanta GA USA Univ Washington Sch Med Dept Internal Med Seattle WA 98195 USA Dana Farber Canc Inst Dept Adult Oncol Boston MA 02115 USA Dana Farber Canc Inst Dept Pediat Oncol Boston MA 02115 USA Harvard Univ Sch Med Boston MA 02115 USA Stanford Univ Sch Med Dept Dermatol Stanford CA 94305 USA Washington Univ Sch Med Dept Neurol St Louis MO 63110 USA Univ Texas Dept Pediat Med Genet Div Pediat Hematol Oncol Houston TX USA

Angiomyolipomas are benign tumors of the kidney derived from putative perivascular epithelioid cells, that may undergo differentiation into cells with features of melanocytes, smooth muscle, and fat. To gain further insight into angiomyolipomas, we have generated the first human angiomyolipoma cell fine by sequential introduction of SV40 large T antigen and human telomerase into human angiomyolipoma cells. These cells show phenotypic characteristics of angiomyolipomas, namely differentiation markers of smooth muscle (smooth muscle actin), adipose tissue (peroxisome proliferator-activator receptor gamma, PPAR gamma), and melanocytes (microophthalmia, MITF), thus demonstrating that a single cell type can exhibit all of these phenotypes. These cells should serve as a valuable tool to elucidate signal transduction pathways underlying renal angiomyolipomas.

关键词：肌动蛋白类/分析脂肪组织/细胞学脂肪组织/病理学血管肌脂瘤/遗传学血管肌脂瘤/病理学血管肌脂瘤/超微结构抗原多瘤病毒转化/遗传学细胞培养技术/方法免疫组织化学肾肿瘤/遗传学肾肿瘤/病理学肾肿瘤/超微结构黑素细胞/细胞学黑素细胞/病理学丝裂原激活蛋白激酶类/分析肌平滑/细胞学肌平滑/病理学表型磷酰化蛋白质类/分析蛋白质类/遗传学受体胞质和核/分析受体胞质和核/代谢阻遏蛋白质类/分析阻遏蛋白质类/遗传学猴病毒40/遗传学端粒末端转移酶/分析端粒末端转移酶/代谢转录因子/分析转录因子/代谢转染肿瘤细胞培养的肿瘤抑制蛋白质类人类

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Modifications in the phosphoinositide signaling pathway by adrenal glucocorticoids in rat brain: Focus on phosphoinositide-specific phospholipase C and inositol 1,4,5-trisphosphate

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第1期295卷 244-254页

作者： Dwivedi, Y Rizavi, HS Rao, JS Pandey, GN Univ Illinois Dept Psychiat MC 912 Inst Psychiat Coll Med Chicago IL 60612 USA

The hypothalamic-pituitary-adrenal (HPA) axis has been shown to be involved in mood and behavior. The possibility that adrenal glucocorticoids regulate components of the phosphatidylinositol (PI) signal transduction pathway was investigated. Two different doses of corticosterone (CORT) pellets (50 or 100 mg) were implanted in normal and bilaterally adrenalectomized (ADX) rats, and CORT regulation of the expression of G(q) alpha protein, phospholipase C (PLC) isozymes, inositol 1,4,5-trisphosphate receptor (IP3R) isoforms, and of PI-PLC activity, [H-3]IP3 binding to IP(3)Rs, and IP3 levels were measured in various brain areas after 1 or 14 days. Fourteen days of CORT pellet implantation into normal rats dose dependently decreased PI-PLC activity and selectively the mRNA and protein expression of PLC beta 1 isozyme in cortex and hippocampus. Bilateral ADX caused the opposite changes in these measures, and simultaneous CORT pellet implantation into ADX rats reversed these effects. Furthermore, 14 days of CORT treatment of normal rats increased [H-3]IP3 binding to IP(3)Rs and decreased IP3 levels in cortex, hippocampus, and cerebellum, without any changes in expression of IP3R-I, IP3R-II, or IP3R-III isoform. On the other hand, ADX decreased [H-3]IP3 binding and increased levels of IP3, and simultaneous CORT treatment of ADX rats prevented these changes. ADX or CORT treatment had no significant effects on the expression of G(q/11) alpha protein. These results suggest that manipulation of the HPA axis alters various components of the PI signaling pathway in rat brain, which may have physiological relevance to the HPA axis-mediated changes in mood and behavior.

关键词：肾上腺切除术钙通道/分析皮质酮/血液皮质酮/药理学肌醇1 4 5-三磷酸/分析肌醇1 4 5-三磷酸/生理学同工酶类/生理学磷脂酰肌醇二酰甘油裂合酶 RNA 信使/分析大鼠 Sprague-Dawley 受体胞质和核/分析动物男(雄)性大鼠

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Determination of complement of mRNA encoding inositol 1,4,5-trisphosphate receptor isoforms in rat tissues using relative-polymerase chain reaction

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ANALYTICAL BIOCHEMISTRY 2001年第2期294卷 188-190页

作者： Gersting, JA Bidasee, KR Dinçer, ÜD Besch, HR Indiana Univ Sch Med Dept Pharmacol & Toxicol Indianapolis IN 46202 USA Ankara Univ Fac Pharm Dept Pharm TR-06100 Ankara Turkey

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Sex-dependent regulation of hepatic peroxisome proliferation in mice by trichloroethylene via peroxisome proliferator-activated receptor α (PPARα)

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CARCINOGENESIS 2000年第4期21卷 677-682页

作者： Nakajima, T Kamijo, Y Usuda, N Liang, Y Fukushima, Y Kametani, K Gonzalez, FJ Aoyama, T Shinshu Univ Sch Med Dept Hyg Matsumoto Nagano 3908621 Japan Shinshu Univ Sch Med Dept Internal Med 2 Matsumoto Nagano 3908621 Japan Shinshu Univ Sch Med Dept Anat 1 Matsumoto Nagano 3908621 Japan Shinshu Univ Sch Med Dept Aging Biochem Matsumoto Nagano 3908621 Japan Shinshu Univ Sch Med Gen Res Lab Matsumoto Nagano 3908621 Japan NCI Lab Metab Bethesda MD 20892 USA

The mechanism of trichloroethylene-induced liver peroxisome proliferation and the sex difference in response was investigated using wild-type Sv/129 and peroxisome proliferator-activated receptor alpha (PPAR alpha)-null mice. Trichloroethylene treatment (0.75 g/kg for 2 weeks by gavage) resulted in liver peroxisome proliferation in wild-type mice, but not in PPAR alpha-null mice, suggesting that trichloroethylene-induced peroxisome proliferation is primarily mediated by PPAR alpha. No remarkable sex difference was observed in induction of peroxisome proliferation, as measured morphologically, but a markedly higher induction of several enzymes and PPAR alpha protein and mRNA was found in males. On the other hand, trichloroethylene induced liver cytochrome P450 2E1, the principal enzyme responsible for metabolizing trichloroethylene to chloral hydrate, only in males, which resulted in similar expression levels in both sexes after the treatment. Trichloroethylene influenced neither the level of catalase, an enzyme involved in the reduction of oxidative stress, nor aldehyde dehydrogenase, the main enzyme catalyzing the conversion to trichloroacetic acid. These results suggest that trichloroethylene treatment causes a male-specific PPAR alpha-dependent increase in cellular oxidative stress.

关键词：细胞分裂/药物作用肝/药物作用肝/超微结构微体/药物作用微体/超微结构受体胞质和核/分析受体胞质和核/药物作用受体胞质和核/生理学受体维甲酸/分析维甲类X受体性别因素转录因子/分析转录因子/药物作用转录因子/生理学三氯乙烯/代谢三氯乙烯/毒性动物女(雌)性男(雄)性小鼠

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Receptor activator NF-κB ligand (RANKL) expression in synovial tissue from patients with rheumatoid arthritis, spondyloarthropathy, osteoarthritis, and from normal patients:: semiquantitative and quantitative analysis

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ANNALS OF THE RHEUMATIC DISEASES 2002年第12期61卷 1047-1054页

作者： Crotti, TN Smith, MD Weedon, H Ahern, MJ Findlay, DM Kraan, M Tak, PP Haynes, DR Univ Adelaide Dept Pathol Adelaide SA 5005 Australia Flinders Med Ctr Dept Med Adelaide SA Australia Repatriat Gen Hosp Rheumatol Res Unit Adelaide SA Australia Flinders Univ S Australia Dept Med Adelaide SA 5001 Australia Univ Adelaide Dept Orthopaed Adelaide SA 5005 Australia Amsterdam Med Ctr Amsterdam Netherlands

Objectives: To compare receptor activator of NF-kappaB ligand (RANKL) production in the synovial tissue from patients with active rheumatoid arthritis (RA), inactive RA, spondyloarthropathies (SpA), osteoarthritis, and from normal subjects. In addition, to establish the cell lineages expressing RANKL in these tissues. Methods: Immunohistological analysis of frozen synovial tissue biopsy specimens was performed using a monoclonal antibody (mAb) to detect RANKL. Sections were evaluated by computer assisted image analysis and semiquantitative analysis to compare RANKL expression between groups. Dual and sequential labelling with mAb RANKL and cell lineage specific monoclonal antibodies were used to determine the types of cells expressing RANKL. Results: Higher levels of RANKL were expressed in tissues from patients with active RA and SpA than in tissues from patients with inactive RA, osteoarthritis, and from normal subjects. RANKL protein was associated with CD3 antigen-positive lymphocytes and some macrophages. RANKL was predominantly associated with activated, memory T cells (CD45Ro positive cells) in patients with active RA and spondyloarthropathy (SpA). Conclusions: The highest levels of RANKL were detected in patients with RA with active synovitis and in some patients with SpA. An increase in RANKL in the inflamed joint of patients with RA, produced by infiltrating activated T cells and macrophages, is likely to be an important cause of joint erosions in RA.

关键词：关节炎类风湿/代谢糖蛋白类/分析免疫酶技术白细胞单核/代谢淋巴细胞/免疫学巨噬细胞/免疫学骨关节炎/代谢骨保护素受体胞质和核/分析受体肿瘤坏死因子/分析椎关节病变/代谢滑液/代谢成年人老年人女(雌)性人类男(雄)性中年人

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Peroxisome proliferator-activated receptor α is restricted to hepatic parenchymal cells, not Kupffer cells:: implications for the mechanism of action of peroxisome proliferators in hepatocarcinogenesis

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CARCINOGENESIS 2000年第4期21卷 823-826页

作者： Peters, JM Rusyn, I Rose, ML Gonzalez, FJ Thurman, RG NCI Lab Metab NIH Bethesda MD 20892 USA Univ N Carolina Dept Pharmacol Hepatobiol & Toxicol Lab Chapel Hill NC 27599 USA

Peroxisome proliferators increase hepatocyte proliferation and cause liver tumors in rodents, yet the mechanism of action is not understood. Based on studies with null mice it is known that peroxisome proliferator-activated receptor-alpha (PPAR alpha) is involved. There is also evidence that Kupffer cells play a central role in peroxisome proliferator-induced carcinogenesis, most likely via mechanisms involving increases in superoxide, activation of nuclear factor kappa B and production of tumor necrosis factor-alpha (TNF alpha), However, it is not known whether PPAR alpha is constitutively expressed in Kupffer cells. Therefore, the expression of PPAR isoforms in rat Kupffer and parenchymal cells was examined. Kupffer cells and hepatocytes of >99 % purity mere isolated from rats fed either a control diet or one containing 0.1% WY-14,643 for 1 week. Protein and RNA were obtained and PPAR expression was analyzed using northern and western blots. PPAR alpha, PPAR beta and PPAR gamma mRNA was detected in purified hepatocytes, In Kupffer cells, mRNA encoding PPAR gamma was present while transcripts for PPAR alpha and PPAR beta were not detected, Immunoblots were consistent with the results found by northern analysis. Moreover, when Kupffer cells from wild-type or PPAR alpha-null mice were treated with WY-14,643 in vitro, superoxide production was similar. Combined, these results show that PPAR alpha is expressed in rat parenchymal cells but not in Kupffer cells. These data are consistent with the hypothesis that parenchymal cells respond to Kupffer cell-derived TNF alpha via mechanisms dependent on PPARa within the parenchymal cells.

关键词：致癌物/毒性枯否细胞/化学肝/化学肝肿瘤实验性/病因学过氧化酶体激增剂/毒性 RNA 信使/分析大鼠 Sprague-Dawley 受体胞质和核/分析受体胞质和核/遗传学受体胞质和核/生理学转录因子/分析转录因子/遗传学转录因子/生理学肿瘤坏死因子α/生理学动物女(雌)性大鼠

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