Obesity is associated with dysbiosis and a state of chronic inflammation that contributes to the pathogenesis of metabolic diseases, including diabetes. We have previously shown that obese mice develop glucose intolerance, increased alloreactivity, and accelerated transplant rejection. In the present study, we investigated the influence of the microbiota on diet-induced obesity (DIO)-associated transplant rejection and hyperglycemia. Antibiotic treatment prolonged graft survival and reduced fasting glycemia in high-fat diet (HFD)-fed specific-pathogen-free (SPF) mice, supporting a role for the microbiota in promoting accelerated graft rejection and hyperglycemia induced by DIO. Further supporting a microbiota-dependent effect, fecal microbiota transfer from DIO SPF mice into germ-free mice also accelerated graft rejection when compared with lean mice-fecal microbiota transfer. Notably, HFD could be also detrimental to the graft independently from microbiota, obesity, and hyperglycemia. Thus, whereas HFD-associated hyperglycemia was exclusively microbiota-dependent, HFD affected transplant outcomes via both microbiota-dependent and -independent mechanisms. Importantly, hyperglycemia in DIO SPF mice could be reduced by the addition of the gut commensal Alistipes onderdonkii, which alleviated both HFD-induced inflammation and glucose intolerance. Thus, microbial dysbiosis can be manipulated via antibiotics or select probiotics to counter some of the pathogenic effects of obesity in transplantation.

Background Nonalcoholic fatty liver disease (NAFLD) is a global health problem. Identifying early gene indicators contributing to the onset and progression of NAFLD has the potential to develop novel targets for early therapeutic intervention. We report on the early and late transcriptomic signatures of western diet (WD)-induced nonalcoholic steatohepatitis (NASH) in female and male Ldlr−/− mice, with time-points at 1 week and 40 weeks on the WD. Control Ldlr−/− mice were maintained on a low-fat diet (LFD) for 1 and 40 weeks. Methods The approach included quantitation of anthropometric and hepatic histology markers of disease as well as the hepatic transcriptome. Results Only mice fed the WD for 40 weeks revealed evidence of NASH, i.e., hepatic steatosis and fibrosis. RNASeq transcriptome analysis, however, revealed multiple cell-specific changes in gene expression after 1 week that persisted to 40 weeks on the WD. These early markers of disease include induction of acute phase response (Saa1-2, Orm2), fibrosis (Col1A1, Col1A2, TGFβ) and NASH associated macrophage (NAM, i.e., Trem2 high, Mmp12 low). We also noted the induction of transcripts associated with metabolic syndrome, including Mmp12, Trem2, Gpnmb, Lgals3 and Lpl. Finally, 1 week of WD feeding was sufficient to significantly induce TNFα, a cytokine involved in both hepatic and systemic inflammation. Conclusion This study revealed early onset changes in the hepatic transcriptome that develop well before any anthropometric or histological evidence of NALFD or NASH and pointed to cell-specific targeting for the prevention of disease progression.

Intestinal commensals can exert immunomodulatory effects on the host, with beneficial or detrimental consequences depending on underlying diseases. We have previously correlated longer survival of minor mismatched skin grafts in mice with the presence of an intestinal commensal bacterium, Alistipes onderdonkii. In this study, we investigated its sufficiency and mechanism of action. Oral administration of A onderdonkii strain DSM19147 but not DSM108265 was sufficient to prolong minor mismatched skin graft survival through inhibition of tumor necrosis factor production. Through metabolomic and metagenomic comparisons between DSM19147 and DSM108265, we identified candidate gene products associated with the anti-inflammatory effect of DSM19147. A onderdonkii DSM19147 can lower inflammation both at a steady state and after transplantation and may serve as an anti-inflammatory probiotic beneficial for transplant recipients.

Abstract Clinical and preclinical studies established that supplementing diets with ω3 polyunsaturated fatty acids (PUFA) can reduce hepatic dysfunction in nonalcoholic steatohepatitis (NASH) but molecular underpinnings of this action were elusive. Herein, we used multi‐omic network analysis that unveiled critical molecular pathways involved in ω3 PUFA effects in a preclinical mouse model of western diet induced NASH. Since NASH is a precursor of liver cancer, we also performed meta‐analysis of human liver cancer transcriptomes that uncovered betacellulin as a key EGFR‐binding protein upregulated in liver cancer and downregulated by ω3 PUFAs in animals and humans with NASH. We then confirmed that betacellulin acts by promoting proliferation of quiescent hepatic stellate cells, inducing transforming growth factor–β2 and increasing collagen production. When used in combination with TLR2/4 agonists, betacellulin upregulated integrins in macrophages thereby potentiating inflammation and fibrosis. Taken together, our results suggest that suppression of betacellulin is one of the key mechanisms associated with anti‐inflammatory and anti‐fibrotic effects of ω3 PUFA on NASH.

The gut microbiota contributes to macrophage-mediated inflammation in adipose tissue with consumption of an obesogenic diet, thus driving the development of metabolic syndrome. There is a need to identify and develop interventions that abrogate this condition. The hops-derived prenylated flavonoid xanthohumol (XN) and its semi-synthetic derivative tetrahydroxanthohumol (TXN) attenuate high-fat diet-induced obesity, hepatosteatosis and metabolic syndrome in C57Bl/6J mice. This coincides with a decrease in pro-inflammatory gene expression in the gut and adipose tissue, together with alterations in the gut microbiota and bile acid composition. In this study, we integrated and interrogated multi-omics data from different organs with fecal 16S sequences and systemic metabolic phenotypic data using a transkingdom network analysis. By incorporating cell type information from single cell RNA-seq data, we discovered TXN attenuates macrophage inflammatory processes in adipose tissue. TXN treatment also reversed levels of inflammation-inducing microbes, such as Oscillibacter valericigenes , that lead to adverse metabolic phenotypes. Furthermore, in vitro validation in macrophage cell lines and in vivo mouse supplementation showed addition of O. valericigenes supernatant induced the expression of metabolic macrophage signature genes that are downregulated by TXN in vivo . Our findings establish an important mechanism by which TXN mitigates adverse phenotypic outcomes from diet-induced obesity and metabolic syndrome. It primarily reduces the abundance of pro-inflammatory gut microbes that can otherwise promote macrophage-associated inflammation in adipose tissue.

>The prevalence of type 2 diabetes(T2D) has escalated to a global epidemic,garnering widespread concern.Tremendous research utilizing germ-free(GF) mice,antibiotic treatments,and fecal microbiota transplantation approaches has established the critical role of gut microbiota in modulating glucose metabolism.The microbiota interacts with diets and the mucosal immune system,influencing intestinal permeability and the penetration of microbial products.

Solid organ transplantation is the preferred treatment for end-stage organ failure. Although transplant recipients take life-long immunosuppressive drugs, a substantial percentage of them still reject their allografts. Strikingly, barrier organs colonized with microbiota have significantly shorter half-lives than non-barrier transplanted organs, even in immunosuppressed hosts. We previously demonstrated that skin allografts monocolonized with the common human commensal Staphylococcus epidermidis (S.epi) are rejected faster than germ-free (GF) allografts in mice because the presence of S.epi augments the effector alloimmune response locally in the graft. Here, we tested whether host immune responses against graft-resident commensal microbes, including S.epi, can damage colonized grafts independently from the alloresponse. Naive hosts mounted an anticommensal T cell response to colonized, but not GF, syngeneic skin grafts. Whereas naive antigraft commensal T cells modestly damaged colonized syngeneic skin grafts, hosts with prior anticommensal T cell memory mounted a post-transplant immune response against graft-resident commensals that significantly damaged colonized, syngeneic skin grafts. Importantly, allograft recipients harboring this host-versus-commensal immune response resisted immunosuppression. The dual effects of host-versus-commensal and host-versus-allograft responses may partially explain why colonized organs have poorer outcomes than sterile organs in the clinic.

Microbiota contribute to the induction of type 2 diabetes by high-fat/high-sugar (HFHS) diet, but which organs/pathways are impacted by microbiota remain unknown. Using multiorgan network and transkingdom analyses, we found that microbiota-dependent impairment of OXPHOS/mitochondria in white adipose tissue (WAT) plays a primary role in regulating systemic glucose metabolism. The follow-up analysis established that Mmp12+ macrophages link microbiota-dependent inflammation and OXPHOS damage in WAT. Moreover, the molecular signature of Mmp12+ macrophages in WAT was associated with insulin resistance in obese patients. Next, we tested the functional effects of MMP12 and found that Mmp12 genetic deficiency or MMP12 inhibition improved glucose metabolism in conventional, but not in germ-free mice. MMP12 treatment induced insulin resistance in adipocytes. TLR2-ligands present in Oscillibacter valericigenes bacteria, which are expanded by HFHS, induce Mmp12 in WAT macrophages in a MYD88-ATF3-dependent manner. Thus, HFHS induces Mmp12+ macrophages and MMP12, representing a microbiota-dependent bridge between inflammation and mitochondrial damage in WAT and causing insulin resistance.

Helicobacter pylori (H. pylori) is one of the most prevalent pathogens globally, and long-term infection causes various gastrointestinal diseases such as gastritis and even cancer. In the present study, we screened dozens of lactic acid bacteria for the efficacy to inhibit H. pylori growth in vitro, and tested the therapeutic effects of candidate strains in vivo. The results showed that Limosilactobacillus fermentum MN-LF23 (LF23) and Lactobacillus gasseri MN-LG80 (LG80) significantly reduced the abundance of Helicobacter by 90% and 83% in the infected mice, respectively, and decreased the levels of serum urease and H. pylori-specific IgG. Both bacterial strains tended to ameliorate H. pylori infection-induced gastric mucosa damage and lymphocyte infiltration, and reduced levels of serum inflammatory cytokines such as TNF-α, IL-1β, and IL-6. In addition, their culture supernatants also showed a therapeutic effect, as efficient as the bacterial cells. Furthermore, both strains significantly regulated gastric microbiota profile, and their supernatants restored the diversity of gastric microbiota. LF23 increased the abundance of Lactobacillus murinus and reduced the abundance of Desulfovibrio, whereas LG80 increased the abundance of Lactobacillus reuteri and reduced the abundance of Bilophila. Both LF23 and LG80 enriched beneficial commensals such as Faecalibaculum rodentium, and reduced detrimental bacteria such as H. pylori and Lachnoclostridium. In conclusion, we identified two novel lactic acid bacteria L. fermentum MN-LF23 and L. gasseri MN-LG80 that can remarkably inhibit H. pylori infection.

Western diet (WD) is one of the major culprits of metabolic disease including type 2 diabetes (T2D) with gut microbiota playing an important role in modulating effects of the diet. Herein, we use a data-driven approach (Transkingdom Network analysis) to model host-microbiome interactions under WD to infer which members of microbiota contribute to the altered host metabolism. Interrogation of this network pointed to taxa with potential beneficial or harmful effects on host’s metabolism. We then validate the functional role of the predicted bacteria in regulating metabolism and show that they act via different host pathways. Our gene expression and electron microscopy studies show that two species from Lactobacillus genus act upon mitochondria in the liver leading to the improvement of lipid metabolism. Metabolomics analyses revealed that reduced glutathione may mediate these effects. Our study identifies potential probiotic strains for T2D and provides important insights into mechanisms of their action.

The microbiome is an environmental factor in intricate symbiotic relationship with its hosts' immune system, potentially shaping anticancer immunity, autoimmunity, and transplant responses. The focus of this review is to discuss recent findings tying the microbiota to transplant outcomes and alloimmunity. The microbiota changes dynamically following transplantation, but whether these changes affect transplant outcomes can be difficult to parse out. New data reveal effects of the microbiota locally, as well as systemically, depending on the mucosal/epithelial surface colonized, the specific commensal communities present and the nature of microbial-derived molecules produced. These complex interactions result in the microbiota potentially impacting transplantation at different levels, including modulation of donor and/or recipient cells, alterations in the priming and/or effector phases of the alloimmune response, availability or metabolism of immunosuppressive drugs, transplant fate or post-transplant complications.

Salicornia herbacea (glasswort) is a traditional Asian medicinal plant which exhibits multiple nutraceutical and pharmaceutical properties. Quercetin-3-glucoside and isorhamnetin-3-glucoside are the major flavonoid glycosides found in S. herbacea. Multiple researchers have shown that flavonoid glycosides can be structurally transformed into minor aglycone molecules, which play a significant role in exerting physiological responses in vivo. However, minor aglycone molecule levels in S. herbacea are very low. In this study, Bifidobacterium animalis subsp. lactis AD011, isolated from infant feces, catalyzed >85% of quercetin-3-glucoside and isorhamnetin-3-glucoside into quercetin and isorhamnetin, respectively, in 2 h, without breaking down flavonoid backbones. Functionality analysis demonstrated that the quercetin and isorhamnetin produced showed improved anti-inflammatory activity vs. the original source molecules against lipopolysaccharide induced RAW 264.7 macrophages. Our report highlights a novel protocol for rapid quercetin and isorhamnetin production from S. herbacea flavonoids and the applicability of quercetin and isorhamnetin as nutraceutical molecules with enhanced anti-inflammatory properties.

A substantial body of literature has provided evidence for the role of gut microbiota in metabolic diseases including type 2 diabetes. However, reports vary regarding the association of particular taxonomic groups with disease. In this systematic review, we focused on the potential role of different bacterial taxa affecting diabetes. We have summarized evidence from 42 human studies reporting microbial associations with disease, and have identified supporting preclinical studies or clinical trials using treatments with probiotics. Among the commonly reported findings, the genera of Bifidobacterium, Bacteroides, Faecalibacterium, Akkermansia and Roseburia were negatively associated with T2D, while the genera of Ruminococcus, Fusobacterium, and Blautia were positively associated with T2D. We also discussed potential molecular mechanisms of microbiota effects in the onset and progression of T2D.

该研究探讨应用表达重组抗表皮生长因子受体三型变异体(EGFRvⅢ)的单链可变区片段(scFv)抗体的重组腺病毒感染骨肉瘤细胞,靶向抑制EGFRvⅢ活性后,骨肉瘤恶性生物学行为的变化情况.收获重组腺病毒,感染骨肉瘤细胞143B、MG63、TE85,靶向抑制EGFRvⅢ活性;体外通过MTT、结晶紫染色、划痕实验、细胞吸附、细胞Matrigel迁移等实验探讨骨肉瘤细胞增殖、黏附、迁移、侵袭等肿瘤恶性生物学行为的变化情况.体内通过感染骨肉瘤动物模型,靶向抑制EGFRvⅢ活性后,应用Xenogen imaging成像技术分析对肿瘤生长的影响.体外实验结果示,靶向抑制骨肉瘤EGFRvⅢ活性,可以明显下调肿瘤细胞的增殖、黏附、迁移、侵袭能力,逆转肿瘤恶性生物学行为;体内实验结果示,靶向抑制骨肉瘤EGFRvⅢ活性,可明显下调肿瘤细胞的增殖生长能力.靶向抑制EGFRvⅢ活性可能逆转骨肉瘤恶性生物学行为,其可能成为骨肉瘤治疗中极有希望的新靶点.

目的 探讨在胆汁淤积性肝硬化病理微环境下对肝脏干细胞存活率的影响及体外甘氨鹅脱氧胆酸钠(GCDC)对肝脏干细胞凋亡的影响.方法 以Balb/c小鼠胆总管结扎模型模拟胆汁淤积性肝硬化的病理环境并回输肝脏干细胞HP14-19,检测细胞定植存活情况;通过细胞计数试剂盒8(CCK-8)、结晶紫染色检测甘氨鹅脱氧胆酸钠对增殖的影响,流式细胞术检测细胞凋亡,通过Western blotting检测凋亡标志物Bax、Bcl-2、Caspase-3表达和磷酸化腺嘌呤核苷二磷酸依赖性蛋白激酶α(p-AMPKα)、腺嘌呤核苷二磷酸依赖性蛋白激酶α(AMPKα)等的表达.结果 通过CCK-8检测和结晶紫染色发现,随着甘氨鹅脱氧胆酸钠作用浓度的增加,HP14-19细胞的增殖抑制;流式细胞术提示,GCDC处理组细胞凋亡率明显增加(P＜0.05);Western blotting检测发现,与对照组相比,实验组凋亡基因Bax、Caspase-3蛋白表达上调,抗凋亡基因Bcl-2蛋白表达下调,AMPKα活化增加(P＜0.05).结论 胆汁淤积性肝硬化所导致的胆汁淤积微环境诱导肝脏干细胞发生凋亡.

The aim of this study is to investigate the changes of cytokines derived from pathologic microenvironment of cholestatic cirrhosis in different time points of common bile duct ligation mices, and to find the optimal combination of cytokines to induce liver stem cells HP14-19 efficiently to differentiate into mature hepatocytes in vitro. The Balb/c mices underwent choledochal ligation (BDL) to simulate the microenvironment of cholestatic cirrhosis, the levels of EGF, HGF and TGF-a in liver tissue of mice with common bile duct ligation detected by immunohistochemistry. The mouse embryonic liver stem cells HP14-19 cells were employed in this study. ALB-Gluc assay was performed to evaluate ALB synthesis ability at different concentrations and different time points. qRT-PCR and Western-blot were used to detect the expression of differentiated cell markers AFP, CK18, ALB. ICG uptake and PAS staining were carried out to detect the metabolism and synthesis function of induced HP14-19 cells. Mouse choledochal ligation can successfully simulate cholestatic cirrhosis, and increase the degree of cirrhosis with the ligation time. Compared with the control, the activity of ALB-Gluc in HP14-19 cells was enhanced after treatment of EGF (10 ng/mL), HGF (20 ng/mL) and TGF-a (20 ng/mL) alone, and markedly enhanced in EGF (10 ng/mL), HGF (20 ng/mL), TGF-a (20 ng/mL) in combined treatment. The results of qRT-PCR and Western blot showed that ALB and CK18 expression increased as the growth of the time and AFP level was opposite, and ICG uptake and PAS staining were significantly increased. The results suggested that cholestasis of liver cirrhosis pathological microenvironment-derived cytokines could effectively promote the differentiation of liver stem cells into mature hepatocytes, and it might have a certain guiding role in the treatment of cholestatic cirrhosis through cytokines combined with liver stem cell transplantation.

Ginseng and probiotics have anti-obesity effects in mice fed a high-fat diet (HFD). Absorption of ginsenoside and colonization of probiotics occur in the intestine. In this study, a mixture of fermented ginseng and two probiotics, Bifidobacterium longum BORI and Lactobacillus paracasei CH88, was administered to HFD-fed mice for 9 weeks. The mixture significantly suppressed weight gain (p < 0.05, n = 8) and lipid deposition in the liver and adipose tissues as well as increased the mice's food intake. The adipocyte size of the adipose tissue was significantly decreased in the mixture-fed group, especially when 0.5% fermented ginseng and 5 × 10⁸/ml of the two probiotics were used (p < 0.05, n = 10). The expression of TNF-α in adipose tissue was efficiently downregulated in the mixture-fed group (p < 0.05, n = 4). The supplement also improved the mice's fasting blood glucose levels (p < 0.05, n = 8) and total cholesterol feces excretion (p < 0.05, n = 8). The mixture of fermented ginseng and B. longum BORI and L. paracasei CH88 could have an anti-obesity effect and suppress lipid deposit in the liver and adipose tissues.

OBJECTIVE:To investigate the changes of tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) in mice with cholestatic cirrhosis and their role in regulating the balance of liver stem cell differentiation.METHODS:Balb/c mice were subjected to bile duct ligation (BDL), and serum biochemical parameters were measured and hepatic histopathology was observed using HE staining to evaluate the modeling of cholestatic cirrhosis. Immunohistochemistry and Western blotting were used to detect the changes of TNF-α and TGF-β1 in the mice after modeling. Mouse embryonic hepatic stem cells (HP14-19) were treated with different concentrations of TNF-α and TGF-β1, and the cell differentiation was assessed using Western blotting, real-time PCR, and PAS staining.RESULTS:The mice receiving BDL showed significantly increased blood biochemical parameters (P<0.05), and HE staining revealed obviously increased collagen fibers in the liver with significantly increased expressions of TNF-α and TGF-β1 (P<0.05). In HP14-19 cells, induction with TNF-α and TGF-β1 for 3 days did not cause significant changes in cell differentiation, but induction for 5 days resulted in significantly increases intensity of PAS staining in the cells. The cells induced with 20, 40, and 80 ng/mL TNF-α for 5 days exhibited a significantly stronger expression of cytokeratin 18 than cytokeratin 19 (P<0.05), while induction with 20, 40, and 80 ng/mL TGF-β1 produced opposite changes in cytokeratin 18 and cytokeratin 19 expressions. Further induction of the cells with TNF-α and TGF-β1 for 10 days, did not alter the expression patterns of cytokeratin 18 and cytokeratin 19 observed on day 5, but their protein expression levels and PAS staining intensity of the cells were enhanced and their mRNA expressions became lowered.CONCLUSION:Common bile duct ligation can induce conditions simulating cholestatic cirrhosis in mice. TNF-α and TGF-β1 are elevated in cholestatic cirrhosis and play opposite roles in regulating the differentiation balance of liver stem cells: the former promotes the differentiation of liver stem cells into hepatocytes, while the latter promotes the cell differentiation into colangiocytes.

Although ginsenosides Rb1, Rb2, Rc, and Rd and ginseng extracts are shown to inhibit pancreatic lipase (PL) activity, the effects of the other ginsenosides, particularly the deglycosylated ones that are considered to show stronger biological activities than the glycosylated forms, are not clear. In this study, we observed the effects of various ginsenosides on PL activity. Results showed that the effects vary with each individual ginsenoside. Ginsenosides Rb1, Rd, Rg1, Rg3, and compound K significantly suppressed 43, 47, 75, and 55% of PL activity at the concentration of 100 μg/mL, respectively. Rg3 was discovered to be the most effective among various common ginsenosides, with a minimum effective concentration of 6.25 μg/mL. Ginsenosides F2 and Rf slightly enhanced PL activity. In addition, fermentation markedly enhanced the inhibitory effect of the ginseng root and ginseng berry, which might be attributed to the changes of ginsenoside profiles.

Korean ginseng was fermented using Aspergillus niger (A. niger) FMB S494 and mycotoxins such as ochratoxin and fumonisin were not detected in the fermented ginseng. Protopanaxadiol-type ginsenosides such as glycosylated forms of Rb1, Rb2, Rc, and Rd decreased to 0 while compound K (cK) increased from 0 to 9 × 104 ppm in the extract of fermented ginseng. Protopanaxtriol-type ginsenosides such as Re and Rg1 decreased from 7.1 × 104 to 3.0 × 104 ppm and 6.8 × 104 to 4.6 × 104 ppm, respectively. Rg2 and Rh1 increased from 0 to 1.9 × 104 ppm and 0 to 2.7 × 104 ppm, respectively. We can demonstrate that A. niger was more inclined to transform protopanaxadiol-type ginsenosides. Moreover, fermented ginseng extract showed a dramatically enhanced anti-proliferative effect on human HT-29 cell line with a minimum effective concentration of about 1 µg/mL, which might be attributed to the high degree of biotransformation of ginsenosides, especially the high output of ginsenoside cK.