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1.
低氧促进大鼠心脏成纤维细胞DNA合成及Ⅰ、Ⅲ胶原mRNA表达 总被引:1,自引:0,他引:1
目的:研究低氧(2%氧)对成年Wistar大鼠心脏成纤维细胞DNA合成及Ⅰ、Ⅲ型胶原前a肽链表达的影响.方法:分离培养成年Wistar大鼠心脏成纤维细胞,采用液体闪烁计数方法检测心脏成纤维细胞的DNA合成速率,采用原位杂交技术检测Ⅰ、Ⅲ型胶原前α肽链mRNA的表达.结果:成年Wistar大鼠心脏成纤维细胞在低氧第6h、12 h时3H-TdR掺入量较常氧组显著增加,分别增加34%(P<0.05)和36%(P<0.01);低氧第4 h、8 h、12 h Ⅰ型胶原前α肽链mRNA表达显著高于常氧培养的细胞;低氧第2 h,Ⅲ型胶原前α肽链mRNA表达显著高于常氧培养的细胞.结论:低氧能够直接促进体外培养的成年Wistar大鼠心脏成纤维细胞DNA合成和Ⅰ、Ⅲ型胶原前α肽链表达,提示低氧对心脏成纤维细胞生长和胶原表达的直接调节可能是低氧性心肌纤维化的重要机制. 相似文献
2.
Jan E. M. Souren Maria Ponec Roeland van Wijk 《In vitro cellular & developmental biology. Plant》1989,25(11):1039-1045
Summary In the process of wound healing keratinocytes and fibroblasts play an important role, keratinocytes in the re-epithelization
process and fibroblasts in the process of wound contraction. We have studied the role of human keratinocytes and fibroblasts
in the rearrangement of collagen in a collagen lattice model system. Our results revealed that keratinocytes as well as fibroblasts
rearrange the collagen lattice; this occurs in a cell number and collagen concentration dependent manner. The optimal gel
contraction is obtained in the presence of keratinocytes on the top of and of fibroblasts in the collagen lattice, the situation
most closely approaching the in vivo situation. Between the two types of cells, differences in morphologic behavior were observed:
when incorporated into the gel the keratinocytes retained their spherical shape throughout the whole culture period, but fibroblasts
became elongated and formed extensions. Our data suggest that not only fibroblasts but also keratinocytes may be involved
in the process of wound contraction.
This work was supported by the Koningin Wilhelmina Fonds (Netherlands Cancer Foundation, grant 84-10). 相似文献
3.
Ruth S. Quinn Stephen M. Krane 《Biochimica et Biophysica Acta (BBA)/General Subjects》1979,585(4):589-598
It has been previously shown that dermis from subjects with hydroxylysine-deficient collagen contains approximately 5% of normal levels of hydroxylysine and sonicates of skin fibroblasts contain less than 15% of normal levels of collagen lysyl hydroxylase activity. However, cultures of dermal fibroblasts from two siblings with hydroxylysine-deficient collagen (Ehlers-Danlos Syndrome Type VI) compared to fibroblasts from normal subjects synthesize collagen containing approximately 50% of normal amounts of hydroxylysine. The lysyl hydroxylase deficient cultures synthesize both Type I and Type III collagen in the same proportion as control cultures. Both α1(I) and α2 chains are similarly reduced in hydroxylysine content. Collagen prolyl hydroxylation by normal and mutant cells is severely depressed without ascorbate but in all cultures collagen lysyl hydroxylation is the same with or without ascorbate supplementation. In mutant cells the rate of prolyl hydroxylation measured after release of inhibition by α,α′-dipyridyl is the same as in control cells. The rate of lysyl hydroxylation is reduced in mutant cells but only to approximately 50% of normal. 相似文献
4.
Fibroblasts cultivated in tridimensional collagen lattices exhibit a downregulation of protein synthesis, related to decreased ribosomal RNA (rRNA) content and half life, when compared to monolayer cultivated cells. The involvement in this process of nucleophosmin/B23, a nucleolar phosphoprotein with ribonuclease properties, was checked. We compared production of nucleophosmin/B23 in monolayer and collagen lattice cultured fibroblasts. A significant increase of nucleophosmin/B23 mRNA levels was noticed in lattice-cultured fibroblasts vs monolayers (+154%, p < 0.05). A concomitant enhancement of nucleolar nucleophosmin/B23 content was found (+112%, p < 0.001). Simultaneously, ribonuclease activity contained in nucleolar extracts from collagen lattice-cultured fibroblasts was significantly increased (+54%, p < 0.01). These data demonstrate that extracellular collagen matrix induces the overexpression of nucleophosmin/B23, and suggest that the regulation of protein syntheses in collagen lattice cultures may be explained, at least partly, by an increased degradation of neosynthesized rRNAs dependent on nucleophosmin. 相似文献
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Kuruvilla L Nair RR Umashankar PR Lal AV Kartha CC 《Cell biochemistry and biophysics》2007,47(1):65-72
Given that vascular endothelial cells play an important role in the modulation of vascular structure and function, we hypothesized
that endocardial endothelial cells (EECs) may have a modulator role in regulating the cardiac interstitial cells. Endocardial
endothelial cells were isolated from freshly collected pig hearts and cardiac fibroblasts were isolated from 3- to 4-d-old
Wistar rats. Fibroblasts were cultured in the presence or absence of conditioned medium from EECs. Proliferation of cardiac
fibroblasts was measured by the incorporation of [3H]-Thymidine and collagen synthesis was assayed by the incorporation of [3H]-proline. To determine the involvement of signaling mediators, in separate experiments, cardiac fibroblasts were incubated
with BQ123 (selective ETA receptor antagonist), PD142893 (nonselective ETA/ETB receptor antagonist), Bis-indolylmaleimide (PKC inhibitor), PD 098059 (MEK inhibitor), or neutralizing anti-transforming
growth factor (TGF)-β-antibody. Endocardial endothelium-derived factors endothelin (ET)-1, TGF-β, and Angiotensin (Ang)-II
in the conditioned medium were assayed by enzyme-linked immunosorbent assay using commercially available kits. We report here
evidence that suggest that endocardial endothelial cells stimulate both proliferation and collagen synthesis of cardiac fibroblasts.
The response seems to be mediated by endothelin through its ETA receptor.
Our results also indicate that protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) pathways are essential for
the EEC-induced proliferation of cardiac fibroblasts. 相似文献
7.
The present study examined the effect of Cerium on protein synthesis in cultured cardiac myocytes and lung fibroblasts exposed to normal and markedly subnormal levels of Mg2+. Cerium was found to have a general inhibitory effect on protein synthesis in these cell types, including the synthesis of myofibrillar proteins in the cardiac myocytes. Further, the effect of the metal ion was more pronounced in cells exposed to the Mg2+-deficient medium. The possible implications of the observations are discussed. 相似文献
8.
Chick embryo sternal chondrocytes from the caudal and cephalic regions were cultured within type I collagen gels and type I collagen/proteoglycan aggregate composite gels in normal serum. Caudal region chondrocytes were also cultured within type I collagen gels in the presence of fibronectindepleted serum. There was a marked stimulation of type X collagen synthesis by the caudal region chondrocytes after 9 days in the presence of fibronectin-depleted serum and after 14 days in the presence of proteoglycan aggregate. These results provide evidence for the ability of chondrocytes from a zone of permanent cartilage to synthesise type X collagen and for the involvement of extracellular matrix components in the control of type X collagen gene expression. 相似文献
9.
Teruo Nishida Atsuko Ueda Masahiko Fukuda Hiroshi Mishima Kyoko Yasumoto Toshifumi Otori 《In vitro cellular & developmental biology. Plant》1988,24(10):1009-1014
Summary Corneal fibroblasts, also known as keratocytes are surrounded by an extracellular matrix of collagen in vivo. To understand
the physiology and pathology of these corneal fibroblasts, it is important to study their interactions with this extracellular
matrix. We cultured rabbit corneal fibroblasts on tissue culture plastic dishes or in a hydrated collagen gel and compared
the changes in morphology and mitotic activity. Corneal fibroblasts on plastic dishes were flattened and widely spread, whereas
those in collagen gel became spindle-shaped with long processes. Examination with an electron microscope revealed that the
corneal fibroblasts in collagen gel formed gap junctions with neighboring cells. Gap junctions were hardly ever observed between
corneal fibroblasts cultured on plastic dishes. Corneal fibroblasts cultured in a collagen matrix showed much less incorporation
of [3H]thymidine than did corneal fibroblasts cultured on plastic, and this incorporation decreased with increasing concentration
of collagen. Our present results suggest that the morphologic and biochemical characteristics of corneal fibroblasts cultured
in collagen gel are different from those cultured on plastic.
This research was supported in part by grants from the Ministry of Education, Science and Culture of Japan, by a grant from
Osaka Eye Bank, Osaka, Japan, and by an intramural research fund of Kinki University.
Part of this research was presented at the annual meeting of the Japanese Ophthalmological Society (May 1985) at Kyoto, Japan,
and at the annual meeting of the Association for Research in Vision and Ophthalmology (May 1987) at Sarasota, FL. 相似文献
10.
Susan H. Kennedy Susan Rouda Huiping Qin Sirpa Aho Jesse Selber Elaine M.L. Tan 《Journal of cellular biochemistry》1997,65(1):32-41
Basic fibroblast growth factor (bFGF) is a mitogenic factor that is implicated in smooth muscle cell growth in atherosclerosis and vascular restenosis. In this study, we examined the effect of bFGF on the expression of the interstitial collagenase gene in human vascular smooth muscle cells. Results from Northern transfer analysis showed that bFGF increased collagenase mRNA levels greater than threefold as early as 24 h. Collagenase pre-mRNA levels were elevated approximately threefold by bFGF, according to RT-PCR analysis. Transient transfections of the smooth muscle cells with a 4.4-kb human collagenase promoter-CAT reporter gene, however, failed to show upregulation of the promoter activity by bFGF. Interestingly, transfections with deleted fragments containing promoter sequences from -1047 to -2271 resulted in modest stimulation of the collagenase-CAT promoter activity by bFGF. bFGF did not alter the stability of the collagenase mRNA, as demonstrated by degradation studies. The enhanced collagenase mRNA levels elicited by bFGF were reflected in increased amounts of collagenase protein that were detected by Western blot analysis. In summary, bFGF upregulates the interstitial collagenase expression, resulting in turnover of the extracellular matrix, an event that could facilitate smooth muscle cell migration and proliferation during the early stages of atherosclerosis and restenosis. J. Cell. Biochem. 65:32–41. © 1997 Wiley-Liss, Inc. 相似文献
11.
R. Edward Branson Kenneth J. Lembach Leon W. Cunningham 《In vitro cellular & developmental biology. Plant》1980,16(2):159-167
Summary Cultured fibroblasts derived from normal subjects and juvenile diabetics attach in the absence of serum to plastic culture
dishes and secrete macromolecules, including collagenous components, hyaluronic acid, and proteoglycans into the medium and
onto the plastic surface where they form a microexudate carpet. Most diabetic fibroblasts examined did not spread as well
as normal cells during a 4-hr interval after the initial attachment. There were no significant differences between normal
and diabetic cells with respect to proline and lysine incorporation and lysine hydroxylation. The percentage glycosylation
of hydroxylysine was marginally higher in the media proteins of diabetic cells, but glycosylation in both normal and diabetic
cells was elevated over that typically observed in human skin collagen.
Collagenous components were estimated to constitute approximately 15–20% of the microexudate carpet fraction in both normal
and diabetic cell strains. Diabetic fibroblasts exhibited a marginally lower ratio of heparan sulfate to chondroitin sulfate
in the cell surface to matrix microexudate carpet fraction (trypsinate) than did normal fibroblasts. The hyaluronate and chondroitin
sulfate contents of this fraction of diabetic cells were not significantly different from those of normal cells.
This work was supported by Grants AM 11821 and AM 19606 from the National Institutes of Health, United States Public Health
Service, by a grant from the American Diabetes Association (Leon W. Cunningham, Principal Investigator) and by the Vanderbilt
Diabetes Endocrinology Center grant, AM 17026. R. E. Branson was the recipient of a research fellowship of the Juvenile Diabetes
Foundation and of a National Institutes of Health Postdoctoral Fellowship AM 05636. 相似文献
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AcSDKP对PDGF诱导的大鼠心脏成纤维细胞增殖和胶原合成的调节作用 总被引:2,自引:0,他引:2
目的:探讨N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酰(AcSDKP)对血小板源性生长因子(PDGF)诱导的大鼠心脏成纤维细胞增殖和胶原合成的调节作用。方法:建立新生大鼠心脏成纤维细胞系;采用四甲基偶氮唑(MTT)法和^3H-TdR掺入法检测心脏成纤维细胞的增殖;采用^3H-脯氨酸掺入法检测心脏成纤维细胞胶原的合成。结果:PD3F在1~20ng/ml浓度范围内对心脏成纤维细胞增殖和胶原合成均有促进作用。且随着PDGF浓度的增加,其促细胞增殖和胶原合成作用增强,并在10ng/ml浓度时PDGF的促增殖和胶原合成效应最强。在10^-10~10^-8mol/L浓度范围内,AcSDKP对PDGF介导的心脏成纤维细胞增殖和胶原合成均有抑制作用,并且在10叫mol/L时,AcSDKP抑制心脏成纤维细胞增殖和胶原合成作用最强。结论:AcSDKP对PDGF介导的心脏成纤维增殖和胶原合成均有明显抑制作用,这可能与其抗心脏纤维化的作用相关。 相似文献
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15.
Fibroblasts cultured from affected skin areas of five patients with cutaneous scleroderma were found to produce increased amounts of collagen when compared with nonaffected control cells. Total RNA was isolated from the cultures and analyzed for its level of pro alpha 1 (I)collagen mRNA by hybridization of RNA blots with a cloned cDNA probe. The levels of pro alpha 1 (I)collagen mRNAs relative to total RNA were two- to sixfold higher in the samples from affected cells, accounting for the increased synthesis of type I collagen. Cytoplasmic dot hybridizations were performed to measure the cellular content of pro alpha 1 (I)collagen mRNA: up to ninefold increases in the level of this mRNA per cell were found. Upon subculturing, scleroderma fibroblasts were found to reduce gradually the increased synthesis of collagen to the level of nonaffected controls by the tenth passage. The levels of type I collagen mRNAs were also reduced, but more slowly. The results suggest that in scleroderma fibroblasts the genes for type I collagen are activated at procollagen mRNA level or that they are more stable and that the activating factors are lost during prolonged cell culture because cells from affected areas lose their activated state. 相似文献
16.
Alain Mauviel Cynthia Halcin Panayiotis Vasiloudes William C. Parks Markku Kurkinen Jouni Uitto 《Journal of cellular biochemistry》1994,54(4):465-472
The degradative effects of interleukin-1 (IL-1) on the extracellular matrix of connective tissue are mediated primarily by metalloproteinases and prostaglandins. Clinical observations suggest that these effects can be prevented, to some extent, by the use of non-steroidal anti-inflammatory drugs. We have examined the role of prostaglandin E2 (PGE2) in IL-1-induced gene expression by human skin fibroblasts in culture. Incubation of confluent fibroblast cultures with varying concentrations (0.01–1.0 μg/ml) of PGE2 led to a dose-dependent elevation of collagenase mRNA steady-state levels, the promoter activity, and the secretion of the protein, whereas relatively little effect was observed on stromelysin and TIMP gene expression. Exogenous PGE2 had no additive or synergistic effect with IL-1 on collagenase gene expression. Furthermore, commonly used non-steroidal anti-inflammatory drugs (indomethacin, acetyl salicylic acid and ibuprofen), at doses which block prostaglandin synthesis in cultured fibroblasts, failed to counteract IL-1-induced collagenase and stromelysin gene expression, nor did they affect TIMP expression. Although the effects of PGE2 did not potentiate those of IL-1 on collagenase gene expression in vitro, one could speculate that massive production of PGE2 by connective tissue cells in vivo in response to inflammatory mediators such as IL-1 or tumor necrosis factor-α, could lead to sustained expression of collagenase in connective tissue cells after clearance of the growth factors. 相似文献
17.
Molecular and Cellular Biochemistry - Recent studies suggest that angiotensin II (angiotensin) may be involved in the regulation of metabolism of the cardiac extracellular matrix (ECM). Two major... 相似文献
18.
Agocha Augustine E. Eghbali-Webb Mahboubeh 《Molecular and cellular biochemistry》1997,172(1-2):195-198
Cardiac fibroblasts constitute greater than 90% of non-myocyte cells in the heart. Because they are responsible for synthesis of components of the extracellular matrix, growth factors and cytokines in the myocardium, they play an important role in normal and pathologic performance of the heart. An understanding of their biology requires in depth studies in a stable and reliable system in which the biological responses of cardiac fibroblasts to various stimuli can be determined. With the exception of few, all studies have been performed on cardiac fibroblasts obtained from rodent hearts. We present a method for isolation and subsequent culture of viable cardiac fibroblasts from ventricular tissue of adult human. This method allows rapid and reliable isolation and subsequent culture of cardiac fibroblasts from adult heart tissue without the need for cumbersome isolation techniques and complex nutrient-enriched and hormone-supplemented culture media for maintenance. 相似文献
19.
目的:通过观察N-乙酰半胱氨酸(NAC)对大鼠心脏成纤维细胞(CFs)增殖和胶原合成的影响,探讨NAC对心脏重构的作用。方法:以培养的新生SD大鼠CFs为实验对象,给予不同浓度的NAC进行干预,48小时后用MTT比色法检测CFs增殖水平,用3H脯氨酸掺入法测定总胶原合成。结果:与对照组相比,不同浓度NAC作用下的CFs增殖水平和3H脯氨酸掺入量均比对照组低,且具有浓度依赖性(p〈0.05)。结论:NAC能够抑制SD大鼠CFs增殖,并降低其胶原合成,因此NAC对心脏的病理性重构可能具有保护作用。 相似文献
20.
目的:通过观察N-乙酰半胱氨酸(NAC)对大鼠心脏成纤维细胞(CFs)增殖和胶原合成的影响,探讨NAC对心脏重构的作用。方法:以培养的新生SD大鼠CFs为实验对象,给予不同浓度的NAC进行干预,48小时后用MTT比色法检测CFs增殖水平,用3H脯氨酸掺入法测定总胶原合成。结果:与对照组相比,不同浓度NAC作用下的CFs增殖水平和3H脯氨酸掺入量均比对照组低,且具有浓度依赖性(p<0.05)。结论:NAC能够抑制SD大鼠CFs增殖,并降低其胶原合成,因此NAC对心脏的病理性重构可能具有保护作用。 相似文献