Multiomics Approaches Identify Biomarkers for BAT Thermogenesis.

Brown adipose tissue (BAT) thermogenesis confers beneficial effects on metabolic diseases such as obesity and type-2 diabetes. Nevertheless, the mechanism and lipid driving the process that evokes this response have not been investigated yet. Here, a multiomics approach of integrative transcriptomics and lipidomics is used to explore the mechanism of regulating thermogenesis in BAT and providing promising lipid biomarkers and biomarker genes for thermogenic activators as antiobesity drugs. Lipidomics analysis demonstrated that a high abundance of glycerophospholipids and sphingolipids was more significant in BAT than in WAT. Enrichment analysis of upregulated DEGs between WAT and BAT screened suggested that the differences were mainly involved in lipid metabolism. Besides, ß3-adrenergic agonist stimulation reduced the levels of TAG and DAG and increased the content of PC, PE, CL, and LPC and expression of genes involved in thermogenesis, fatty acid elongation, and glycerophospholipid metabolism in BAT. In this study, based on interpreting the inherent characterization of BAT as thermogenic tissue through comparison with WAT as fat storage tissue, adrenergic stimulation-induced BAT thermogenesis further identified specific lipid biomarkers (7 TAG species, 10 PC species, 1 LPC species, and 1 CL species) and Elovl3 and Crat gene biomarkers, which may provide targets for combating obesity by boosting BAT thermogenesis.

Probiotic Lactobacillus casei Zhang reduces pro-inflammatory cytokine production and hepatic inflammation in a rat model of acute liver failure.

PURPOSE: In this study, we sought to find the effects and mechanisms of probiotic Lactobacillus casei Zhang (L. casei Zhang) on the pro-inflammatory cytokine production and hepatic inflammatory response in a rat model of acute liver failure induced by lipopolysaccharide (LPS) and d-galactosamine (GalN). METHODS: Male Wistar rats were orally administrated with or without L. casei Zhang for 30 days prior to challenge with LPS and GalN. Dexamethasone administrated group serving as a positive anti-inflammation control. Serum, intestinal and liver samples were collected 8 h after LPS/GalN challenge for histological, molecular and biochemical analysis. RESULTS: LPS/GalN challenge alone resulted in significantly increased production of endotoxin, tumor necrosis factor-α (TNF-α), interleukin-1 beta (IL-1ß) and nitric oxide as compared to the normal control rats. Pretreatment with L. casei Zhang not only reduced these changes, but also attenuated hepatic inflammation as shown by improved histological assessment, decreased myeloperoxidase activity and reduced expression of IL-1ß and inducible nitric oxide synthase in the liver. L. casei Zhang supplementation significantly inhibited LPS/GalN-triggered phosphorylation of ERK, JNK and p-38 MAPK, but increased the expression of TLR2, TLR9 and PPAR-γ. Moreover, L. casei Zhang treatment prevented intestinal injury and modulated the intestinal ecology by increasing the fecal Lactobacillus and Bifidobacterium levels. CONCLUSIONS: Probiotic L. casei Zhang reduces LPS/GalN-induced pro-inflammatory cytokine and hepatic inflammation through modulating the TLR-MAPK-PPAR-γ signaling pathways and intestinal microbiota.

Successive stamen movement in Saxifraga candelabrum is responsive to weather and pollinator visits.

BACKGROUND: Successive stamen movement is a complex plant behavior involving successive uplift of stamens and pollen release, which plays a role in reducing sexual interference, increasing pollen deposition and promoting pollen export. Although reported from several taxa, studies on whether the movement can be influenced by abiotic and biotic factors are scarce. METHODS: In this study, we here for the first time described a pattern of successive stamen movement in Saxifraga candelabrum (Saxifragaceae). We then compared the rates of stamen movement in S. candelabrum under different weather and varying pollinator visits. Pollen packaging and presentation schedule of S. candelabrum were also investigated. RESULTS: The results showed that the number of stamens bent per day in sunny days was significantly higher than overcast and rain. Flowers that receive more pollinator visits (control treatment) had significantly higher number of stamen movement than those that received fewer (removal treatment) and none (bagging treatment). Throughout the staminate phase of a flower, there was a progressive increase in both pollen quantity of individual stamens and pollen presentation during each day. CONCLUSION: Our research demonstrates that successive stamen movement in S. candelabrum was accelerated by favorable weather and increased pollinator visits, which may promote pollen export. Moreover, incremental pollen packaging is likely an adaptation to seasonal regularity in variations of sex ratio resulting from protandry.

Lactobacillus casei Zhang modulate cytokine and toll-like receptor expression and beneficially regulate poly I:C-induced immune responses in RAW264.7 macrophages.

Lactobacilli are frequently used as probiotics due to their beneficial effects on health. Lactobacillus casei Zhang (LcZ), which has favorable probiotic properties, was first isolated from koumiss. In this study, the immunomodulating effects of LcZ on cytokine and toll-like receptor expression in RAW264.7 macrophages was assessed and it was found that live LcZ promotes production of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-6 and interferon (IFN)-ß. Transcription of inducible nitric oxide synthase (iNOS) was also enhanced by viable LcZ. The immunostimulating effects of live LcZ are significantly attenuated in heat-killed LcZ. Live LcZ promotes TLR2 mRNA transcription, whereas heat-killed LcZ enhances transcription of TLR2, TLR3, TLR4 and TLR9. Furthermore, live LcZ significantly suppresses polyinosinic:polycytidylic acid (poly I:C)-stimulated NO, iNOS and TNF-α expression while enhancing expression of IFN-ß. It was also found that poly I:C-induced interferon regulatory factor 3 (IRF-3) reporter gene activity was significantly up-regulated by live LcZ. These results suggest that LcZ keeps the innate immune system alert by increasing transcription of Toll-like receptors and enhancing production of pro-inflammatory mediators and type I IFN in macrophages. The synergistic effect of live LcZ with poly I:C on IFN-ß expression is associated with increased activity of IRF-3. LcZ has the potential to be used as an adjuvant against viral infections.

Inhibition of glycolysis prevents behavioural changes in mice with MK801-induced SCZ model by alleviating lactate accumulation and lactylation.

Schizophrenia (SCZ) is a debilitating neuropsychiatric disorder with a complex aetiology. Cognitive symptoms and hippocampal changes have been implicated in the pathophysiology of SCZ. Changes in metabolites level and up-regulated glycolysis have been reported in previous studies, which may be related to the hippocampal dysfunction in SCZ. However, the pathological mechanism of glycolysis involved in the pathogenesis of SCZ remains unclear. Therefore, the change of glycolysis level and the involvement in SCZ need to be further studied. In our study, MK801 was used to induce an SCZ mouse model and cell model in vivo and in vitro. Western blotting was performed to evaluate the levels of glycolysis, metabolites, and lactylation in hippocampal tissue of mice with SCZ or cell models. The level of high mobility group protein 1 (HMGB1) in the medium of MK801-treated primary hippocampal neurons was examined. Apoptosis was evaluated in HMGB1-treated hippocampal neurons by flow cytometry. The glycolysis inhibitor 2-DG prevented behavioural changes in the MK801-induced SCZ mouse model. The lactate accumulation and level of lactylation were alleviated in the hippocampal tissue of MK801-treated mice. Glycolysis was enhanced, and lactate accumulated in MK-801-treated primary hippocampal neurons. In addition, the level of HMGB1 increased in the medium and induced apoptosis in primary hippocampal neurons. Together, the data showed that glycolysis and lactylation increased in the MK801-induced SCZ model in vivo and in vitro, and this effect could be prevented by 2-DG (a glycolysis inhibitor). Glycolytic related HMGB1 upregulation may induce apoptosis in hippocampal neurons downstream.

Kaempferol Alleviates Murine Experimental Colitis by Restoring Gut Microbiota and Inhibiting the LPS-TLR4-NF-κB Axis.

Intestinal microbiota dysbiosis is an established characteristic of ulcerative colitis (UC). Regulating the gut microbiota is an attractive alternative UC treatment strategy, considering the potential adverse effects of synthetic drugs used to treat UC. Kaempferol (Kae) is an anti-inflammatory and antioxidant flavonoid derived from a variety of medicinal plants. In this study, we determined the efficacy and mechanism of action of Kae as an anti-UC agent in dextran sulfate sodium (DSS)-induced colitis mice. DSS challenge in a mouse model of UC led to weight loss, diarrhea accompanied by mucous and blood, histological abnormalities, and shortening of the colon, all of which were significantly alleviated by pretreatment with Kae. In addition, intestinal permeability was shown to improve using fluorescein isothiocyanate (FITC)-dextran administration. DSS-induced destruction of the intestinal barrier was also significantly prevented by Kae administration via increases in the levels of ZO-1, occludin, and claudin-1. Furthermore, Kae pretreatment decreased the levels of IL-1ß, IL-6, and TNF-α and downregulated transcription of an array of inflammatory signaling molecules, while it increased IL-10 mRNA expression. Notably, Kae reshaped the intestinal microbiome by elevating the Firmicutes to Bacteroidetes ratio; increasing the linear discriminant analysis scores of beneficial bacteria, such as Prevotellaceae and Ruminococcaceae; and reducing the richness of Proteobacteria in DSS-challenged mice. There was also an evident shift in the profile of fecal metabolites in the Kae treatment group. Serum LPS levels and downstream TLR4-NF-κB signaling were downregulated by Kae supplementation. Moreover, fecal microbiota transplantation from Kae-treated mice to the DSS-induced mice confirmed the effects of Kae on modulating the gut microbiota to alleviate UC. Therefore, Kae may exert protective effects against colitis mice through regulating the gut microbiota and TLR4-related signaling pathways. This study demonstrates the anti-UC effects of Kae and its potential therapeutic mechanisms, and offers novel insights into the prevention of inflammatory diseases using natural products.

Ebosin Ameliorates Psoriasis-Like Inflammation of Mice via miR-155 Targeting tnfaip3 on IL-17 Pathway.

Psoriasis is a recurrent autoimmune skin disease with aberrant regulation of keratinocytes and immunocytes. There is no universally accepted single treatment available for psoriasis, and the establishment of a common treatment option to control its signs and symptoms is urgently needed. Here, we found Ebosin, a novel exopolysaccharide isolated from Streptomyces sp. 139 by our lab, not only could ameliorate inflammation in LPS-induced keratinocytes through IKK/NF-kapaB pathway, but also attenuate psoriatic skin lesions and reduce inflammatory factors expression in imiquimod (IMQ)-mediated psoriatic mice. Except for inhibiting the expression of epidermal differentiation related proteins, Ebosin significantly increased the percentage of CD4+Foxp3+CD25+ Tregs and decreased CD4+IL17A+ Th17 cells in psoriatic mice. Furthermore, we demonstrate that Ebosin significantly suppressed the IL-17 signaling pathway via A20 (encoded by tnfaip3) in vivo. As the direct binding of tnfaip3 to miR-155 has been demonstrated by luciferase reporter assay, and Ebosin has been demonstrated to inhibit miR-155 level in vitro and in vivo, our study first indicates that Ebosin reduces inflammation through the miR-155-tnfaip3-IL-17 axis and T cell differentiation in a psoriasis-like model. Thus, we conclude that Ebosin can act as a promising therapeutic candidate for the treatment of psoriasis.

Preparation and evaluation of a Rubropunctatin-loaded liposome anticancer drug carrier.

Rubropunctatin is a naturally occurring constituent of polyketide compounds that has great potential in the development of cancer-assisted chemotherapy. However, it has certain shortcomings such as water insolubility and photo instability that limit its clinical application. In this study, we constructed a Rubropunctatin-loaded liposome (R-Liposome) anticancer drug carrier for the first time. The results indicate that R-Liposome is water soluble, has spherical morphology, great homogeneity and dispersibility with high encapsulation efficiency (EE%, 90 ± 3.5%) and loading rate (LR%, 5.60 ± 2.5%) values. Moreover, the carrier improves the photostability, storage and pH stabilities of Rubropunctatin. The R-Liposome also prolongs the release of Rubropunctatin, enhances the anticancer activity of Rubropunctatin and encourages the mechanism of Rubropunctatin to promote apoptosis. Therefore, liposomal nanoparticles have great potential as drug delivery vehicles of Rubropunctatin for cancer treatment.

mTOR Expression in Hippocampus and Prefrontal Cortex Is Downregulated in a Rat Model of Schizophrenia Induced by Chronic Administration of Ketamine.

Schizophrenia is a severe chronic neuropsychiatric disorder, and it negatively affects individuals' quality of life, but the pathogenesis of schizophrenia remains unclear. This study aimed to explore whether the administration of ketamine in rats causes changes in mTOR (mechanistic/mammalian target of rapamycin) expression in the hippocampus and prefrontal cortex. Ketamine was used to establish an animal model of schizophrenia. Rats were randomly divided into four groups: control group (normal saline), low-dose group (15 mg/kg ketamine), middle-dose group (30 mg/kg ketamine), and high-dose group (60 mg/kg ketamine). The rats were intraperitoneally injected with ketamine or normal saline twice a day (9 AM and 9 PM) for 7 consecutive days. Immunohistochemistry was used to detect mTOR protein expression in the hippocampus and prefrontal cortex from rats at 13 h after the last treatment. Using immunohistochemistry, the expression of the mTOR protein was localized exclusively in the CA3 region of the hippocampus and in the Cg1 region of the prefrontal cortexes. Ketamine at 60 mg/kg decreased the expression of mTOR protein in the brain of rats. Ketamine successfully established a rat model of schizophrenia. This study helps elucidate the mechanisms of ketamine-induced schizophrenia and provides novel insights for drug discovery and development.

Knockdown of Rab7a suppresses the proliferation, migration, and xenograft tumor growth of breast cancer cells.

Breast cancer is a common invasive cancer in women. Ras-related protein Rab-7a (Rab7a) is involved in late endocytic trafficking, while its role in breast cancer is largely unclear. In the present study, we investigated the role of Rab7a in breast cancer. Comparing with adjacent breast tissues, Rab7a expression was increased in breast cancer tissues. Using lentivirus-mediated knockdown strategy, we found that Rab7a silencing inhibited the proliferation and colony formation of MDA-MB-231 cells. Apoptosis and G2 cell cycle arrest were induced in Rab7a knockdown. By contrast, Rab7a suppressed the apoptosis and promoted proliferation and colony formation of MCF-7 cells. The migration of MDA-MB-231 cells was suppressed by Rab7a knockdown. In vivo, depletion of Rab7a inhibited the xenograft tumor development of MDA-MB-231 cells. Altogether, our results highlight the novel function of Rab7a in the proliferation, invasion, and xenograft tumor development of breast cancer cells.