Janus Kinase 2 (JAK2) Dissociates Hepatosteatosis from Hepatocellular Carcinoma in Mice.

Hepatocellular carcinoma is an end-stage complication of non-alcoholic fatty liver disease (NAFLD). Inflammation plays a critical role in the progression of non-alcoholic fatty liver disease and the development of hepatocellular carcinoma. However, whether steatosis per se promotes liver cancer, and the molecular mechanisms that control the progression in this disease spectrum remain largely elusive. The Janus kinase signal transducers and activators of transcription (JAK-STAT) pathway mediates signal transduction by numerous cytokines that regulate inflammation and may contribute to hepatocarcinogenesis. Mice with hepatocyte-specific deletion of JAK2 (L-JAK2 KO) develop extensive fatty liver spontaneously. We show here that this simple steatosis was insufficient to drive carcinogenesis. In fact, L-JAK2 KO mice were markedly protected from chemically induced tumor formation. Using the methionine choline-deficient dietary model to induce steatohepatitis, we found that steatohepatitis development was completely arrested in L-JAK2 KO mice despite the presence of steatosis, suggesting that JAK2 is the critical factor required for inflammatory progression in the liver. In line with this, L-JAK2 KO mice exhibited attenuated inflammation after chemical carcinogen challenge. This was associated with increased hepatocyte apoptosis without elevated compensatory proliferation, thus thwarting expansion of transformed hepatocytes. Taken together, our findings identify an indispensable role of JAK2 in hepatocarcinogenesis through regulating critical inflammatory pathways. Targeting the JAK-STAT pathway may provide a novel therapeutic option for the treatment of hepatocellular carcinoma.

DJ-1 links muscle ROS production with metabolic reprogramming and systemic energy homeostasis in mice.

Reactive oxygen species (ROS) have been linked to a wide variety of pathologies, including obesity and diabetes, but ROS also act as endogenous signalling molecules, regulating numerous biological processes. DJ-1 is one of the most evolutionarily conserved proteins across species, and mutations in DJ-1 have been linked to some cases of Parkinson's disease. Here we show that DJ-1 maintains cellular metabolic homeostasis via modulating ROS levels in murine skeletal muscles, revealing a role of DJ-1 in maintaining efficient fuel utilization. We demonstrate that, in the absence of DJ-1, ROS uncouple mitochondrial respiration and activate AMP-activated protein kinase, which triggers Warburg-like metabolic reprogramming in muscle cells. Accordingly, DJ-1 knockout mice exhibit higher energy expenditure and are protected from obesity, insulin resistance and diabetes in the setting of fuel surplus. Our data suggest that promoting mitochondrial uncoupling may be a potential strategy for the treatment of obesity-associated metabolic disorders.

Redundant role of the cytochrome c-mediated intrinsic apoptotic pathway in pancreatic ß-cells.

Cytochrome c is one of the central mediators of the mitochondrial or the intrinsic apoptotic pathway. Mice harboring a 'knock-in' mutation of cytochrome c, impairing only its apoptotic function, have permitted studies on the essential role of cytochrome c-mediated apoptosis in various tissue homeostasis. To this end, we examined the role of cytochrome c in pancreatic ß-cells under homeostatic conditions and in diabetes models, including those induced by streptozotocin (STZ) and c-Myc. Previous studies have shown that both STZ- and c-Myc-induced ß-cell apoptosis is mediated through caspase-3 activation; however, the precise mechanism in these modes of cell death was not characterized. The results of our study show that lack of functional cytochrome c does not affect glucose homeostasis or pancreatic ß-cell mass under basal conditions. Moreover, the cytochrome c-mediated intrinsic apoptotic pathway is required for neither STZ- nor c-Myc-induced ß-cell death. We also observed that the extrinsic apoptotic pathway mediated through caspase-8 was not essential in c-Myc-induced ß-cell destruction. These findings suggest that cytochrome c is not required for STZ-induced ß-cell apoptosis and, together with the caspase-8-mediated extrinsic pathway, plays a redundant role in c-Myc-induced ß-cell apoptosis.

Embryonic survival and severity of cardiac and craniofacial defects are affected by genetic background in fibroblast growth factor-16 null mice.

Disruption of the X-chromosome fibroblast growth factor 16 (Fgf-16) gene, a member of the FGF-9 subfamily with FGF-20, was linked with an effect on cardiac development in two independent studies. However, poor trabeculation with lethality by embryonic day (E) 11.5 was associated with only one, involving maintenance in Black Swiss (Bsw) versus C57BL/6 mice. The aim of this study was to examine the potential influence of genetic background through breeding the null mutation onto an alternate (C57BL/6) background. After three generations, 25% of Fgf-16(-/Y) mice survived to adulthood, which could be reversed by reducing the contribution of the C57BL/6 genetic background by back crossing to another strain. There was no significant difference between FGF-9 and FGF-20 RNA levels in Fgf-16 null versus wild-type mice regardless of strain. However, FGF-8 RNA levels were reduced significantly in Bsw but not C57BL/6 mice. FGF-8 is linked to anterior heart development and like the FGF-9 subfamily is reportedly expressed at E10.5. Like FGF-16, neuregulin as well as signaling via ErbB2 and ErbB4 receptors have been linked to trabeculae formation and cardiac development around E10.5. Basal neuregulin, ErbB2, and ErbB4 as well as FGF-8, FGF-9, and FGF-16 RNA levels varied in Bsw versus C57BL/6 mice. These data are consistent with the ability of genetic background to modify the phenotype and affect embryonic survival in Fgf-16 null mice.

Estrogen affects the differentiation and function of splenic monocyte-derived dendritic cells from normal rats.

AIM: To explore the mechanism resulting in the preventive effect of estrogen on experimental autoimmune encephalomyelitis (EAE), which is an animal model for multiple sclerosis (MS), and examine if estrogen can affect the immune response in EAE at dendritic cell (DC) level. METHODS: Flow cytometry was used to reveal the surface marker expression. 3H-thymidine incorporation was applied to examine the cellular proliferation. Levels of anti-myelin basic protein (MBP)(68-86) antibody and cytokines were determined by enzyme-linked immunospot and ELISA, respectively. RESULTS: 17beta-estradiol (E2) could dose-dependently accelerate the differentiation process of DCs by up-regulating CD11c, B7-2 and CD40 expressions, but exert no effect on its antigen presentation ability. MBP-specific T cells cocultured with E2-treated DCs (E2-DC) produced more IL-10 and less IFN-gamma in the supernatant than those without E2 pretreatment (ctr-DC). In contrast to ctr-DC, E2-DC, if injected i.v. into EAE rats on day 5 post immunization, could initiate antigen nonspecific hyper-responsivity in T cells in terms of enhanced proliferation and cytokine secretion of mononuclear cells in LN, but suppressed antibody secretion from splenocytes. CONCLUSION: These results suggest that estrogen can affect the differentiation and function of DCs, which leads T cells switching to Th2 secretion. This may account partly for the protective effect of estrogen on EAE.

Morphological observation of immunological alterations in the spleens from estrogen receptor deficient mice.

AIM: To evaluate some immunological parameters in the spleens from estrogen receptor(ER) deficient mice. METHODS: Immunohistochemistry and immunofluorescence analysis were used in the study. RESULTS: Multiple alterations were found in the spleens of ER deficient mice, especially in ERbeta deficient mice, such as increased expression of proinflammatory cytokines and inducible nitric oxide synthase, elevated number of proliferating splenocytes and increased IgM/IgG production. Moreover, The splenocytes lack of ERbeta were resistant towards estrogen's suppression on proliferation. The activation of nuclear factor (NF)-kappaB in ERbeta deficient spleens may account for the above hyperactivity. CONCLUSION: ER, especially ERbeta, plays a key role in mediating the modulation of immune response by estrogen. ERbeta disruption may increase the sensibility towards autoimmune diseases.

Disruption of estrogen receptor beta in mice brain results in pathological alterations resembling Alzheimer disease.

AIM: To study the pathological characteristics of the mice with estrogen receptor beta (ER beta) disruption in brain. METHODS: Immunohistochemistry method was applied in the study. RESULTS: beta-Amyloid peptide(A beta (42)) and apolipoprotein E (ApoE) immunoreactive substances were accumulated notably in cortex and limbic structures such as the hippocampus and amygdala in brain, resembling the pathological changes of human Alzheimer disease(AD). A beta formed cloudy-like deposits in parenchyma of brain, while apoE also deposited along or surrounding the blood vessels. CONCLUSIONS: ER beta is crucial to the development of neural degenerative disease, so modulation of A beta metabolism via ER beta signal pathway might be beneficial for AD prevention or therapy.

[Inhibition of moderate hypoxia-induced protein synthesis by vasonatrin peptide in cultured neonatal rat cardiomyocytes].

The present work was to investigate the effects of vasonatrin peptide (VNP) on cardiomyocyte protein synthesis induced by moderate hypoxia. In cultured neonatal rat cardiomyocytes, MTT methods, total protein measurement and (3)H-leucine incorporation were used to calculate the cell number and measure the protein synthesis of cardiomyocytes. Furthermore, radioimmunoassay was undertaken to observe the effects of VNP on the intracellular levels of cAMP, cGMP and the concentration of endothelin (ET) in the culture medium. The results showed that both the cell number and protein synthesis decreased with severe hypoxia for 24 h. In contrast, under moderate hypoxia, cardiomyocyte hypertrophy developed; the protein synthesis as evidenced by total protein content and 3H-eucine incorporation increased significantly. VNP reduced cardiomyocyte protein synthesis induced by moderate hypoxia in a dose-dependent manner. Furthermore, VNP increased the intracellular level of cGMP and decreased the concentration of ET in the culture medium under moderate hypoxia, but had no effect on the level of cAMP. These results suggest that VNP inhibits moderate hypoxia-induced protein synthesis in cultured neonatal rat cardiac myocytes. This effect is mediated, at least in part, by an increase in intracellular cGMP, a reduction in synthesis, and/or a release in ET of cardiomyocytes.