Volume 124, Issue 5, Pages (May 2003)

Slides:



Advertisements
Similar presentations
Proinflammatory cytokine–induced and chemical mediator–induced IL-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein.
Advertisements

Chronic exposure of human mesangial cells to high glucose environments activates the p38 MAPK pathway  William A. Wilmer, Cynthia L. Dixon, Courtney Hebert 
Volume 56, Issue 5, Pages (November 1999)
Volume 68, Issue 4, Pages (October 2005)
Volume 127, Issue 5, Pages (November 2004)
Volume 124, Issue 5, Pages (May 2003)
Volume 114, Issue 4, Pages (April 1998)
Volume 127, Issue 5, Pages (November 2004)
Volume 133, Issue 6, Pages (December 2007)
Volume 9, Issue 5, Pages (November 1998)
Volume 56, Issue 4, Pages (October 1999)
Lipopolysaccharide Activates Caspase-1 (Interleukin-1–Converting Enzyme) in Cultured Monocytic and Endothelial Cells by Ralf R. Schumann, Claus Belka,
Constitutive Phosphorylation of Focal Adhesion Kinase Is Involved in the Myofibroblast Differentiation of Scleroderma Fibroblasts  Yoshihiro Mimura, Hironobu.
Volume 129, Issue 1, Pages (July 2005)
Differential influence of tyrosine residues of the common receptor β subunit on multiple signals induced by human GM-CSF  Tohru Itoh, PhD, Rui Liu, MSc,
Volume 127, Issue 3, Pages (September 2004)
Richard T. Ethridge, Mark R. Hellmich, Raymond N. DuBois, B.Mark Evers 
Volume 127, Issue 5, Pages (November 2004)
Prolyl Hydroxylase-3 Is Down-regulated in Colorectal Cancer Cells and Inhibits IKKβ Independent of Hydroxylase Activity  Jing Xue, Xuebing Li, Shi Jiao,
Shitao Li, Lingyan Wang, Michael A. Berman, Ye Zhang, Martin E. Dorf 
Volume 124, Issue 5, Pages (May 2003)
Enteroinvasive bacteria alter barrier and transport properties of human intestinal epithelium: Role of iNOS and COX-2  Silvia Resta–Lenert, Kim E. Barrett 
Autocrine and paracrine functions of vascular endothelial growth factor (VEGF) in renal tubular epithelial cells  Guillermo Villegas, Bäerbel Lange-Sperandio,
John F. Öhd, Katarina Wikström, Anita Sjölander  Gastroenterology 
Volume 114, Issue 4, Pages (April 1998)
Human osteoarthritic chondrocytes are impaired in matrix metalloproteinase-13 inhibition by IFN-γ due to reduced IFN-γ receptor levels  R. Ahmad, M. El.
Volume 141, Issue 4, Pages e2 (October 2011)
Volume 56, Issue 5, Pages (November 1999)
Pim-1 is up-regulated by constitutively activated FLT3 and plays a role in FLT3-mediated cell survival by Kyu-Tae Kim, Kristin Baird, Joon-Young Ahn, Paul.
Andreea M. Bujor, Jaspreet Pannu, Shizhong Bu, Edwin A. Smith, Robin C
Volume 123, Issue 6, Pages (December 2002)
Volume 123, Issue 1, Pages (July 2002)
IGF-II-Mediated COX-2 Gene Expression in Human Keratinocytes Through Extracellular Signal-Regulated Kinase Pathway  Hye Jung Kim, Tae-Yoon Kim  Journal.
Volume 128, Issue 5, Pages (May 2005)
Expression of inter-α-trypsin inhibitor and tumor necrosis factor-stimulated gene 6 in renal proximal tubular epithelial cells  Ulf Janssen, Gareth Thomas,
Volume 118, Issue 6, Pages (June 2000)
STAT3 attenuates EGFR-mediated ERK activation and cell survival during oxidant stress in mouse proximal tubular cells  I. Arany, J.K. Megyesi, B.D. Nelkin,
Volume 23, Issue 2, Pages (July 2006)
Volume 136, Issue 3, Pages e2 (March 2009)
Mechanisms of cross hyporesponsiveness to toll-like receptor bacterial ligands in intestinal epithelial cells  Jan-Michel Otte, Elke Cario, Daniel K.
Volume 68, Issue 4, Pages (October 2005)
Arachidonic acid induces ERK activation via Src SH2 domain association with the epidermal growth factor receptor  L.D. Alexander, Y. Ding, S. Alagarsamy,
Volume 93, Issue 7, Pages (June 1998)
1,25-dihydroxyvitamin D3 inhibits renal interstitial myofibroblast activation by inducing hepatocyte growth factor expression  Yingjian Li, Bradley C.
Volume 132, Issue 5, Pages (May 2007)
Volume 4, Issue 2, Pages (February 1996)
Volume 7, Issue 5, Pages (November 1997)
Chronic exposure of human mesangial cells to high glucose environments activates the p38 MAPK pathway  William A. Wilmer, Cynthia L. Dixon, Courtney Hebert 
Upregulation of Tenascin-C Expression by IL-13 in Human Dermal Fibroblasts via the Phosphoinositide 3-kinase/Akt and the Protein Kinase C Signaling Pathways 
Volume 1, Issue 4, Pages (June 2007)
Volume 124, Issue 7, Pages (June 2003)
DNA binding of activator protein-1 is increased in human mesangial cells cultured in high glucose concentrations  William A. Wilmer, Fernando G. Cosio 
Interleukin-6-Resistant Melanoma Cells Exhibit Reduced Activation of STAT3 and Lack of Inhibition of Cyclin E-Associated Kinase Activity  Markus Böhm,
Volume 63, Issue 6, Pages (June 2003)
Volume 61, Issue 6, Pages (June 2002)
Human Keratinocytes Respond to Osmotic Stress by p38 Map Kinase Regulated Induction of HSP70 and HSP27  M. Garmyn, A. Pupe  Journal of Investigative Dermatology 
Rsk1 mediates a MEK–MAP kinase cell survival signal
Volume 22, Issue 1, Pages (January 2005)
Small heat shock protein alteration provides a mechanism to reduce mesangial cell contractility in diabetes and oxidative stress  Marjorie E. Dunlop,
Mesalamine blocks tumor necrosis factor growth inhibition and nuclear factor κB activation in mouse colonocytes  Greg C. Kaiser, Fang Yan, D.Brent Polk 
Volume 57, Issue 2, Pages (October 2000)
Volume 60, Issue 3, Pages (September 2001)
Lu-Cheng Cao, Thomas Honeyman, Julie Jonassen, Cheryl Scheid 
Volume 55, Issue 2, Pages (February 1999)
Phosphorylation of cytosolic phospholipase A2 by IL-3 is associated with increased free arachidonic acid generation and leukotriene C4 release in human.
Volume 4, Issue 4, Pages (October 1999)
Deon G. Uffort, Elizabeth A. Grimm, Julie A. Ellerhorst 
Unassembled Ig Heavy Chains Do Not Cycle from BiP In Vivo but Require Light Chains to Trigger Their Release  Marc Vanhove, Young-Kwang Usherwood, Linda.
Volume 114, Issue 1, Pages (January 1998)
The JNK phosphatase M3/6 is inhibited by protein-damaging stress
Presentation transcript:

Volume 124, Issue 5, Pages 1358-1368 (May 2003) Interleukin-11-induced heat shock protein 25 confers intestinal epithelial-specific cytoprotection from oxidant stress  Mark J Ropeleski, Jun Tang, Margaret M Walsh-Reitz, Mark W Musch, Eugene B Chang  Gastroenterology  Volume 124, Issue 5, Pages 1358-1368 (May 2003) DOI: 10.1016/S0016-5085(03)00282-8

Figure 1 IL-11 induces rapid phosphorylation and activation of STAT3 at Tyr705 in IEC-18, YAMC, NIH3T3, and MDCK-HR cells. Serum-deprived cells grown in 0.01% FBS for 12 hours were stimulated with IL-11 (100 ng/mL) for 10–120 minutes. Cells were washed and total cell lysates were prepared as previously outlined. Identical membranes were incubated separately with antibodies specific for (Tyr705) phospho-STAT3 and total STAT3. Rapid phosphorylation of STAT3 occurred within 10 minutes of IL-11 stimulation, indicating that all cells expressed functional IL-11Rα chain (C, 0 minutes unstimulated control; C120′, 120 minutes unstimulated control). The figure is representative of 3 separate experiments with cells from different passages. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 4 IL-11 induces intestinal epithelial-specific induction of immunodetectable hsp25 expression. (A ) IEC-18 and YAMC intestinal epithelial cells were exposed to IL-11 50 ng/mL for 24 hours while 3T3 cells were used as nonepithelial controls and MDCK cells were used as nonintestinal epithelial controls. HS IEC-18 cells were used as positive controls. Analysis by SDS PAGE and Western blotting revealed that only intestinal epithelial cells (IEC-18 and YAMC) responded to IL-11 by increasing hsp25 expression. No effect was seen in 3T3, MDCK cells, or RLE-6TN cells. +, IL-11 50ng/mL for 24 hours; −, unstimulated controls. (B) RLE-6TN pulmonary epithelial cells did not express hsp25 in response to IL-11 despite IL-11R functionality demonstrated by STAT3 phosphorylation. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 2 IL-11 induces immunodetectable hsp25 expression in a time-dependent manner. (A ) IEC-18 cells were exposed to IL-11 50ng/mL for 6–72 hours and 10μg of total cell lysates were analyzed by SDS-PAGE. A significant increase in hsp25 expression was observed by 6 hours. Values represent results from 3 independent experiments using cells from different passages. Heat-shocked (HS) IEC-18 cells (IEC-18 cells heat shocked at 42°C for 23 minutes followed by a 2-hour recovery) were used as positive controls and the constitutively expressed Hsc73, which does not respond to IL-11, was used for normalization of densitometric analysis. No effect of IL-11 on hsp72 expression was observed. (B) A more elaborate time course reveals that no induction of hsp25 by IL-11 is seen prior to 4 hours, unlike HS controls where significant induction appears within 2 hours (see Figure 2A representative of n =3). (C ) A significant increase in hsp25 expression was seen by 6 hours, and was maximal by 24 hours which was used as 100% reference for densitometric analysis. Data are expressed as the mean ± SEM where ∗P < 0.05 and ∗∗∗P < 0.001 with respect to unstimulated controls. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 3 IL-11 induces immunodetectable hsp25 in a dose-dependent manner. IEC-18 cells were exposed to various doses of IL-11 in ng/mL for 24 hours, total cell protein lysates were analyzed by SDS-PAGE and Western blotting. A dose-dependent increase in hsp25 expression was observed while no changes in inducible hsp72 expression were observed. Results were normalized to the constitutively expressed hsc73, which is not affected by IL-11 treatment. HS, IEC-18 cells heat shocked at 42°C for 23 minutes followed by a 2-hour recovery. The image shown here is representative of 3 separate experiments. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 5 Anti-sense to hsp25 eliminates basal, IL-11, and heat-induced hsp25 expression. IEC-18 clones transfected with the pCEP4 expression vector containing the full-length rat cDNA for hsp25 sub-cloned in the reverse direction were selected under hygromycin B for stable anti-sense expression. Clone A4 depicted here was used in further studies to determine hsp25 specific protective effects of IL-11 during oxidant stress. Empty pCEP4 vector controls are also depicted here, where no effect of transfection was observed on hsp25 expression. +, IL-11 50 ng/mL for 24 hours; −, no stimulation; HS, 42°C for 23 minutes followed by a 2-hour recovery. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 6 IL-11 protects intestinal epithelial viability from oxidant stress in 51Cr release assays. Empty vector IEC-18 transfectants and IEC-18 cells expressing stable anti-sense to hsp25 were labeled with 51Cr following a 24-hour exposure to IL-11 50 ng/mL as outlined previously. Cytotoxicity was assayed as a function of percent of 51Cr released after a 1-hour challenge with 0.3–0.6mmol/L of the oxidant monochloramine. Unstimulated empty vector IEC-18 transfectants were used as controls. Data are expressed as the mean ± SEM. ∗∗∗P <0.001. ns, no statistical significance. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 7 (A ) IL-11 fails to protect NIH-3T3 cells from oxidant stress in 51Cr release assays. The effects of IL-11 on 3T3 cell resistance to oxidative stress were examined in 51Cr release assays as described in the Materials and Methods section. Unlike assays with IEC-18 cells, 100ng/mL of IL-11 for 24 hours failed to confer any cytoprotection. Because of the relative resistance to oxidant injury compared with IEC-18 cells, the effects of IL-11 were also examined at 1 and 3 mmol/L doses of monochloramine. (B) IL-11 fails to protect IEC-18 cells at time points before the induction of hsp25. The ability of IL-11 100ng/mL to confer cytoprotection to IEC-18 cells in 51Cr release assays was examined after 2 hours of incubation, at time points that antedate the induction of hsp25. Compared with incubation with IL-11 for 24 hours, no significant reduction in 51Cr release was detected. Data are shown as mean and SEM of 3 experiments. ∗∗∗P <0.001 with respect to IEC-18 cells treated with IL-11 for 2 hours. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)

Figure 8 (A ) IL-11 stimulation leads to the accumulation of hsp25 in Triton ×-100 insoluble fractions. IEC-18 cells were exposed to IL-11 and cell pellets were resuspended in 0.5% Triton ×-100 lysis buffer followed by centrifugation. Insoluble pellets were resuspended in SDS solubilization buffer and 20 μg of each sample were analyzed by SDS PAGE. Hsp25 detection was carried out as described previously. Blot is representative of 4 separate experiments. (B) Heat shock leads to a rapid shift of hsp25 to Triton ×-100 insoluble fractions. IEC-18 cells were heat shocked at 43°C for 30 minutes followed by a recovery period at 37°C. Triton ×-100 insoluble proteins (5μg/lane) were analyzed by SDS-PAGE and Western blotting. (C ) Heat shock induces hsp25 phosphorylation in IEC-18 cells. Cells were heat-shocked for 30 minutes at 43°C and immediately harvested for isoelectric focusing 2-dimensional gel electrophoresis. Samples were run in the first dimension in 4% polyacrylamide tube gels containing 5% ampholytes(pH 3–10) and 55% wt/vol high-grade urea. Samples were electrophoresed in the second dimension on 12.5% gels, transferred to polyvinylidene difluoride membranes, and immunoblotted to detect phosphorylated hsp25 isoforms. a, native form; b, monophosphorylated form; c, diphosphorylated form (n = 2). (D) IL-11 treatment of IEC-18 cells does not lead to hsp25 phosphorylation. IEC-18 cells were treated for 4 and 24 hours before harvesting. Two-dimensional isoelectric focusing gel electrophoresis was carried out as described in Figure 8C. No alteration in hsp25 phosphorylation was detected. a, native form, b, monophosphorylated form; c, diphosphorylated form. Blot is representative of 2 experiments. Gastroenterology 2003 124, 1358-1368DOI: (10.1016/S0016-5085(03)00282-8)