Distinct Roles of Smad2-, Smad3-, and ERK-dependent Pathways in Transforming Growth Factor-β1 Regulation of Pancreatic Stellate Cellular Functions

Ohnishi, Hirohide; Miyata, Tomohiko; Yasuda, Hiroshi; Satoh, Yasuyuki; Hanatsuka, Kazunobu; Kita, Hiroto; Ohashi, Akiyoshi; Tamada, Kiichi; Makita, Noriko; Iiri, Taroh; Ueda, Norihiro; Mashima, Hirosato; Sugano, Kentaro

doi:10.1074/jbc.m309698200

Cited by 102 publications

(97 citation statements)

References 27 publications

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“…15,47,48 The same is true for HSC, where contrasting data describing the influence of TGFb1 on proliferation, ranging from growth inhibition to stimulation of proliferation, have been published, although the majority of publications report a TGFb1-induced growth inhibition in HSC. 44,[49][50][51] These differences may be attributed to different RT-PCR demonstrated that cultivation of immortalized PSC on Matrigel (M) for 7 days resulted in a marked decrease of Col I, TGFb1 and CTGF expression compared to cultivation on tissue culture (TC) plates, whereas aSMA expression was not altered significantly by this cultivation conditions, the housekeeping gene RPL13A served as control (a).…”

Section: Deactivation Of Pancreatic Stellate Cells R Jesnowski Et Almentioning

confidence: 90%

Immortalization of pancreatic stellate cells as an in vitro model of pancreatic fibrosis: deactivation is induced by matrigel and N-acetylcysteine

Jesnowski

Fürst

Ringel

et al. 2005

Laboratory Investigation

132

134

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Tissue fibrosis is one of the characteristics of chronic pancreatitis and pancreatic adenocarcinoma. Activated pancreatic stellate cells (PSC) play a central role in this process. However, analysis of the molecular mechanisms leading to PSC activation is hampered by the lack of an established human PSC line. To overcome this problem, we immortalized and characterized primary human PSC. The cells were isolated by the outgrowth method and were immortalized by transfection with SV40 large T antigen and human telomerase (hTERT). Primary human PSC served as controls. An immortalized line, RLT-PSC, was analyzed for the expression of stellate cell markers. Moreover, the effects of transforming growth factor b 1(TGFb1) or platelet-derived growth factor stimulation and of cultivation on basement membrane components or N-acetylcysteine (NAC) treatment on gene and protein expression and proliferation were analyzed. Immortal RLT-PSC cells retained the phenotype of activated PSC proven by the expression of a-smooth muscle actin (aSMA), vimentin, desmin and glial fibrillary acidic protein (GFAP). TGFb1 treatment upregulated the expression of aSMA, collagen type I (Col I), fibronectin and TGFb1. Incubation of RLT-PSC cells and primary human activated PSC on Matrigel plus NAC treatment resulted in a deactivated phenotype as evidenced by a decrease of aSMA, connective tissue growth factor and Col I expression and by a decreased proliferation of the cells. Moreover, this treatment restored the ability of the cells to store vitamin A in cytoplasmic vesicles. In conclusion, we have established an immortal pancreatic stellate cell line, without changing the characteristic phenotype. Importantly, we were able to demonstrate that besides soluble factors, the matrix surrounding PSC plays a pivotal role in the maintenance of the activation process of PSC. Cultivation of activated PSC on a reconstituted basement membrane plus treatment with NAC was able to deactivate the cells, thus pointing to the possibility of an antifibrosis therapy in chronic pancreatitis.

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Section: Deactivation Of Pancreatic Stellate Cells R Jesnowski Et Almentioning

confidence: 90%

Immortalization of pancreatic stellate cells as an in vitro model of pancreatic fibrosis: deactivation is induced by matrigel and N-acetylcysteine

Jesnowski

Fürst

Ringel

et al. 2005

Laboratory Investigation

132

134

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“…Through gene sequence detection, we found that the TGF-β l , Smad2/3, CycA, and ORC genes of PSC sequences were differentially expressed after co-culture with IkBαM-MSCs. Ohnishi et al (2004) found that TGF-b l promotes the activation of PSCs through the Smad2-dependent pathway and that PSCs infected by the adenovirus with the Smad2-dominant-negative gene inhibit PSC activation. IkBαM-MSCs can reduce the secretion of TGF-b 1 , thereby inhibiting the transduction of the TGF-b l -smad-2/3 signaling pathway.…”

Section: Mensenchymal Stell Cells In Chronic Pancreatitismentioning

confidence: 99%

Effect of the IkBα mutant gene delivery to mesenchymal stem cells on rat chronic pancreatitis

et al. 2014

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ABSTRACT. This study aimed to investigate the effect of inhibitors of the NF-kΒ alpha mutant gene (IkBaM) delivery to mensenchymal stem cells (MSCs) on rat chronic pancreatitis (CP). A total of 120 SpragueDawley rats were randomly divided into 6 groups of 20: Group A was injected with sterile saline solution, Group B was injected with allogenic MSCs, Group C1 was injected with allogenic IkBαM-MSCs cultured in vitro 4 h before CP modeling, Group C2 was injected with allogenic IkBαM-MSCs cultured in vitro during CP modeling, Group C3 was cultured with allogenic IkBαM-MSCs cultured in vitro 4 h after CP modeling, and Group D was injected with rAAV2-MSCs. Cytokine levels of ICAM-1, CTGF, IL-1, IL-6, IL-8, TNF-α, TIMP-1, TIMP-2, IL-10, FN, MMP-1, MMP-2, MMP-3, and MMP-9 were examined. The results indicated that allogenic IκBαM-MSCs could reduce pro- inflammatory cytokine levels and increase anti-inflammatory cytokine levels in CP. The allogenic IkBαM-MSCs reduced the activation and promoted the apoptosis of pancreatic stellate cells in the rat model of CP. IkBαM-MSCs influenced the proliferation and apoptosis of pancreatic stellate cells by regulating the activation of the PPAR, MAPK, mTOR, TGF-β, NOD-like receptor, Notch, WNT, TGF-β1-SMAD-2/3, and P53 signal transduction pathways.

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“…Multiple studies have identified several major signaling pathways involved in the regulation of PaSC function (27,43,47,(50)(51)(52). MAPKs are key mediators of activating signals initiated by growth factors, angiotensin II, and ethanol (51,52).…”

Section: Mediators Of Pasc Activationmentioning

confidence: 99%

The pancreatic stellate cell: a star on the rise in pancreatic diseases

Omary¹,

Lugea²,

Lowe³

et al. 2007

J. Clin. Invest.

612

614

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Distinct Roles of Smad2-, Smad3-, and ERK-dependent Pathways in Transforming Growth Factor-β1 Regulation of Pancreatic Stellate Cellular Functions

Cited by 102 publications

References 27 publications

Immortalization of pancreatic stellate cells as an in vitro model of pancreatic fibrosis: deactivation is induced by matrigel and N-acetylcysteine

Immortalization of pancreatic stellate cells as an in vitro model of pancreatic fibrosis: deactivation is induced by matrigel and N-acetylcysteine

Effect of the IkBα mutant gene delivery to mesenchymal stem cells on rat chronic pancreatitis

The pancreatic stellate cell: a star on the rise in pancreatic diseases

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