High mobility group box-1 promotes the proliferation and migration of hepatic stellate cells via TLR4-dependent signal pathways of PI3K/Akt and JNK
Authors: Lei Li
Fu-ping Wang
Ji-yao Wang
Wei Jiang
Jing Li
Ling-yan Wang
Journal: PLoS ONE (2013)
Discipline: Multidisciplinary
Keywords: Hepatic stellate cells; HMGB1; migration; proliferation; TLR4
Affiliations: Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai, China.

Abstract:  Background: The migration of hepatic stellate cells (HSCs) is essential to the hepatic fibrotic response, and recently High-mobility group box 1 (HMGB1) has been shown up-regulated during liver fibrosis. Nevertheless, whether HMGB1 can modulate the proliferation and migration of HSCs is poorly understood, as well as the involved intracellular signaling. In this study, we examined the effect of HMGB1 on proliferation, migration, pro-fibrotic function of HSCs and investigated whether toll-like family of receptor 4 (TLR4) dependent signal pathway is involved in the intracellular signaling regulation. Methodology/principal findings: Modified transwell chamber system to mimic the space of Disse was used to evaluate the migration of human primary HSCs, and the protein expressions of related signal factors were evaluated by western blot. Cell proliferation was analyzed by MTT assay, the pro-fibrotic functions of HSCs by qRT-PCR and ELISA respectively. Recombinant human HMGB1 could significantly promote migration of HSCs under both haptotactic and chemotactic stimulation, especially the latter. Human TLR4 neutralizing antibody could markedly inhibit HMGB1-induced migration of HSCs. HMGB1 could enhance the phosphorylation of JNK and PI3K/Akt, and TLR4 neutralizing antibody inhibited HMGB1-enhanced phosphorylation of JNK and PI3K/Akt and activation of NF-Œ∫B. JNK inhibitor (SP600125) and PI3K inhibitor (LY 294002) significantly inhibited HMGB1-induced proliferation and migration of HSCs, and also reduced HMGB1-enhanced related collagen expressions and pro-fibrotic cytokines production. Conclusions/significance: HMGB1 could significantly enhance migration of HSCs in vitro, and TLR4-dependent JNK and PI3K/Akt signal pathways are involved in the HMGB1-induced proliferation, migration and pro-fibrotic effects of HSCs, which indicates HMGB1 might be an effective target to treat liver fibrosis. 

Retracted: Yes

PubPeer comments:
 2019 retraction. https: journals.plos.org plosone article?id=10.1371 journal.pone.0216942 After publication of this article [1], concerns were raised about results reported in Figs 2 and 3: Data in lanes 1, 2 of the p-Pl3K blot in Fig 2A appear similar to data in lanes 2, 3 of the PI3K blot in Fig 2B. Similarities were noted between bands in lanes 1, 2 of the PI3K blot in Fig 2B. The p-JNK western blot in Fig 2A appears similar to the NF-kB blot in Fig 3A. The authors commented that errors had been made in preparing Figs 2B and 3A, and that the panels shown in Fig 2A are correct. The authors provided a replacement image for the PI3K panel in Fig 2 and replication data for the results shown in Figs 2B and 3A. The data supplied do not address the concerns identified. A member of PLOS ONEu2019s Editorial Board advised that the replication data do not report consistent results, and do not suffice to support the articleu2019s conclusions regarding the effects of HMGB1 and anti-TLR4 on PI3K and JNK signaling and NF-kB activation. Original uncropped image data underlying the published results are not available. In light of the above concerns, the PLOS ONE Editors retract this article. All authors agreed with the retraction. Reference 1. Wang F-p, Li L, Li J, Wang J-y, Wang L-y, Jiang W (2013) High Mobility Group Box-1 Promotes the Proliferation and Migration of Hepatic Stellate Cells via TLR4-Dependent Signal Pathways of PI3K Akt and JNK. PLoS ONE 8(5): e64373. https: doi.org 10.1371 journal.pone.0064373 pmid:23696886 , 
 Here are the similarities between lanes in Figures 2A, 2B, and 3A as described in the retraction notice. In addition, lanes 1 and 2 in the IkBa panel in Figure 3A look similar to each other (shown with green arrows).  ,