Connective tissue growth factor reacts as an IL - World Journal of ...

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A 3 d 5-HT 10 μmol/L 1 min by pretreatment with 1 μmol/L U73122, a phospholipase C inhibitor (0.05 ± 0.05 peak changes of Fura-2 ratio from 0.66 ± 0.12, n = 13). We also observed that [Ca 2+ ]i transients induced by 5-HT were not altered in extracellular Ca 2+ -free conditions (data not shown). These results suggest that 5-HT activates phospholipase C to produce IP3, which induces Ca 2+ release from ER in activated HSCs. To confirm the receptor subtype, we tested blocking effects of a universal 5-HT2 antagonist, ritanserin, which does not discriminate among 5HT2 isotypes. 5-HTinduced [Ca 2+ ]i responses were attenuated by pretreatment WJG|www.wjgnet.com Fura-2 ratio 0.2 5-HT 10 μmol/L 2 wk 1 min D E 500 bp 100 bp B M GAPDH 1A (211) 1B (258) 2A (376) 2B (352) 3A (352) 3B (428) 4 (377) 5A (109) 5B (132) 6 (129) 7 (147) (predicted product size) Relative expression ratio (5-HT2/GAPDH) Fura-2 ratio 0.4 0.08 0.06 0.04 0.02 0.00 5-HT 5-HT2A 5-HT2B 1 min 100 μmol/L 10 μmol/L 3 μmol/L 1 μmol/L Figure 2 5-hydroxytryptamine-induced intracellular Ca 2+ concentration changes and the expression of 5-hydroxytryptamine2 receptors in quiescent and activated hepatic stellate cells. A, B: 5-hydroxytryptamine (5-HT)-induced intracellular Ca 2+ concentration ([Ca 2+ ]i) transients were recorded from hepatic stellate cells (HSCs) at 3 d (A) and 2 wk (B) after isolation; C: Averages of [Ca 2+ ]i changes (from 13-40 cells/each trace) in response to 5-HT (1-100 μmol/L) application to 2 wkcultured HSCs are shown; D: Steady-state mRNA levels of the 5-HT receptor isotypes in 2 wk-cultured HSCs were compared using reverse transcription-polymerase chain reaction (RT-PCR); E: Using quantitative RT-PCR, the transcriptional changes in 5-HT2 receptors among 1 d-, 1 wk- and 2 wk-cultured HSCs were compared. Expression levels were normalized to GAPDH and expressed as a relative expression ratio (target/GAPDH, n = 3). Data are presented as the mean ± SE. A ATP 100 μmol/L 5-HT 10 μmol/L 1 min Fura-2 ratio (F340/F380) 0.2 B Δ Fura-2 ratio (F340/F380) 0.6 0.4 0.2 0.0 Park KS et al . Serotonergic signaling in HSCs (28) C (28) (16) ATP 5-HT ACh Figure 3 Comparison of intracellular Ca 2+ concentration responses to various metabotropic receptor agonists in activated hepatic stellate cells. Intracellular Ca 2+ concentration changes following application of ATP (100 μmol/L), 5-hydroxytryptamine (5-HT) (10 μmol/L), or acetylcholine (ACh, 10 μmol/L) were measured in 2 wkcultured hepatic stellate cells (n = 3-6, 16-28 cells). Data are presented as the mean ± SE. 1 d 1 wk 2 wk Fura-2 ratio with 10 μmol/L ritanserin by 46.3% (0.34 ± 0.08 from 0.89 ± 0.10, n = 11). Upregulation of calcium transporting and binding proteins in the ER In mammalian cells, there are three major subtypes of the sarcoplasmic/endoplasmic reticulum Ca 2+ ATPase (SERCA1, 2, and 3) which pump Ca 2+ into the ER lumen. We observed SERCA2 to be the dominant subtype in HSCs. SERCA2, especially SERCA2b, is considered to be a house-keeping protein expressed constitutively 169 January 14, 2011|Volume 17|Issue 2| 0.1
A C Park KS et al . Serotonergic signaling in HSCs 5-HT 10 μmol/L 5-HT 10 μmol/L Figure 4 5-hydroxytryptamine-induced intracellular Ca 2+ concentration transients via metabotropic 5-hydroxytryptamine2 receptor in activated hepatic stellate cells. A, B: 5-hydroxytryptamine (5-HT)-induced intracellular Ca 2+ concentration ([Ca 2+ ]i) transients were completely abolished by pretreatment with U73122 (1 μmol/L), a phospholipase C blocker (n = 3, 11 cells); C, D: Ritanserin (10 μmol/L), a 5-HT2 antagonist, inhibited the [Ca 2+ ]i responses to 5-HT in activated hepatic stellate cells (2 wk-cultured cells; n = 3, 13 cells). Data are presented as the mean ± SE. WJG|www.wjgnet.com 5-HT 10 μmol/L U73122 1 μmol/L 5 min 5-HT 10 μmol/L Ritanserin 10 μmol/L Fura-2 ratio 5 min Fura-2 ratio in most kinds of cells; however, in HSCs, the expression of SERCA2 tends to increase during activation. Specifically, the relative expression ratio of SERCA2 (SERCA2/ GAPDH) at 1 d after isolation was 0.058, and increased to 0.106 after 1 wk in culture and 0.164 after 2 wk in culture in vitro (Figure 5A). The expression of SERCA3 was also increased during culture (SERCA3/GAPDH; 0.4 × 10 -3 at 1 d and 6.9 × 10 -3 at 2 wk). Among the three isoforms (types 1 through 3) of the IP3 receptor, the type 1 IP3 receptor was the main subtype expressed in activated HSCs. We observed that the expression of the type 1 IP3 receptor increased by about 7-fold (IP3R 1/GAPDH = 0.037) after 1 wk of culture, and 20-fold (0.100) after 2 wk of culture compared to (0.005) levels 1 d after isolation (Figure 5B). In contrast, the expression level of ryanodine receptors, which are a family of Ca 2+ -releasing channel proteins expressed in the ER, either did not change or was decreased during the activation of HSCs (Figure 5C). We investigated whether Ca 2+ binding chaperones of the ER could be up-regulated following the activation process of HSCs. There were similar increases in the expression levels of calreticulin (calreticulin/GAPDH; from 0.204 at 1 d to 0.655 at 2 wk), calnexin (calnexin/GAP- DH; from 0.240 at 1 d to 0.750 at 2 wk), and calsequestrin in HSCs. In the case of calsequestrin, the expression level in HSCs at 1 d after isolation was undetectable, but was markedly increased (calsequestrin/GAPDH; 0.217) after 2 wk of culturing (Figure 5D). 0.2 0.5 B Δ Fura-2 ratio (F340/F380) D Δ Fura-2 ratio (F340/F380) 0.8 0.6 0.4 0.2 0.0 1.0 0.8 0.6 0.4 0.2 0.0 DISCUSSION 5-HT U73122 + 5-HT 5-HT Ritanserin + 5-HT Trans-differentiation of HSCs is accompanied by marked increases in protein synthesis, including collagen, elastin, and glycoproteins [7] . It is well known that Ca 2+ homeostasis in the ER is critical for the synthesis, folding, and secretion of protein [22,23] . In HSCs, the depletion of ER Ca 2+ stores inhibits protein synthesis and increases intracellular degradation of collagen [34] . Maintaining a high Ca 2+ gradient across the ER membrane (around 1000-fold) is accomplished by active Ca 2+ transport by SERCAs. Among the three different isoforms of SERCAs, SERCA2 is considered to be a house-keeping protein expressed in the ER of most cell types, including HSCs [34] . We observed that SERCA2 was the main isotype in quiescent and activated HSCs (Figure 5A). During activation, the expression of SERCA2 (and also SERCA3) was increased, which likely helped to maintain appropriate ER Ca 2+ concentrations. Chaperone proteins in the ER facilitate the folding of newly synthesized proteins and glycoproteins. In particular, calreticulin and calnexin are important chaperones involved in a “quality control” system for protein synthesis [35] . In addition, these chaperones act as Ca 2+ binding proteins in the ER. Overexpression of calreticulin increases the total amount of Ca 2+ in intracellular stores, whereas calreticulin-deficient cells have reduced ER Ca 2+ storage capacity [36] . Impaired collagen synthesis has been observed in cells derived from mice possessing genetic defects in ER chaperone proteins [37] . In this study, we observed for 170 January 14, 2011|Volume 17|Issue 2|
Page 1 and 2: World Journal of Gastroenterology W
Page 3 and 4: Robert JL Fraser, Daw Park Jacob Ge
Page 5 and 6: Italy Donato F Altomare, Bari Piero
Page 7 and 8: Fernando Azpiroz, Barcelona Ramon B
Page 9 and 10: S Contents EDITORIAL TOPIC HIGHLIGH
Page 11 and 12: Contents APPENDIX FLYLEAF EDITORS F
Page 13 and 14: Ouyang D et al . Pancreatic endocri
Page 19 and 20: Online Submissions: http://www.wjgn
Page 21 and 22: Diamantis G et al . Quality of life
Page 27: Gressner OA et al . CTGF is a negat
Page 30 and 31: A B CTGF luciferase activity (lcps)
Page 32 and 33: A B IL-6 (35 μg/L) - - + + + + - s
Page 34 and 35: F mt distal mt proximal wt 9 8 7 6
Page 36 and 37: + CTGF TGF-β IL-6 + - + an acute p
Page 38 and 39: 2007; 46: 295-303 21 Karger A, Fitz
Page 40 and 41: 2011; 17(2): 164-173 Available from
Page 43: A B C -80 mV E Relative expression
Page 48 and 49: 19 Li T, Weng SG, Leng XS, Peng JR,
Page 50 and 51: Recurrent GB cancer has been report
Page 52 and 53: Table 2 Surgical procedures for muc
Page 54 and 55: esection of the common bile duct sh
Page 58 and 59: NaF, 1% Triton X-100, and 0.5 mmol/
Page 60 and 61: A Oxaliplatin LY294002 C Oxaliplati
Page 62 and 63: A Tumor volume (mm 3 ) C 5000 4000
Page 64 and 65: 2 Hundahl SA, Phillips JL, Menck HR
Page 68 and 69: the typical MII pattern of GOR. Thi
Page 70 and 71: emptying, that arises from the anch
Page 74 and 75: medical treatment, age at diagnosis
Page 76 and 77: Table 3 Odds ratio for the studied
Page 78 and 79: Table 6 Published associations betw
Page 80 and 81: anti-CBir1 and ASCA, and show reduc
Page 85: Faria GR et al . Acute diverticulit
Page 90 and 91: Changes in flow vs T0 (%) Changes i
Page 93 and 94: Hanssen SJ et al . Hemolysis and in
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Kuiper P et al . Angiogenic markers
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Kuiper P et al . Angiogenic markers
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A Comulative survival C Comulative
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Online Submissions: http://www.wjgn
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Nishida U et al . ASA induced small
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Nishida U et al . ASA induced small
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Okano K et al . FDG-PET small pancr
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Okano K et al . FDG-PET small pancr
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Table 3 Prevalence of IgG anti-hepa
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1005-1022 24 Hendrickx G, Van Herck
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Choi YS et al . Alternative cleansi
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Plasma endotoxin (EU/L) Small intes
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Zhang Y et al . Effects of penehycl
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C Review: CYP1A1 Ile462Val polymorp
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Kim SO et al . Sorafenib-induced sp
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Sahebkar A. Ginger as a natural sup
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Instructions to authors Indexed and
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Instructions to authors uniform leg
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Instructions to authors Language ev
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