Gut Microbiota Alteration Influences Colorectal Cancer Metastasis to the Liver by Remodeling the Liver Immune Microenvironment

Background/Aims@#This study aimed to explore the effect of gut microbiota-regulated Kupffer cells (KCs) on colorectal cancer (CRC) liver metastasis. @*Methods@#A series of in vivo and in vitro researches were showed to demonstrate the gut microbiota and its possible mechanism in CRC liver metastasis. @*Results@#Fewer liver metastases were identified in the ampicillin-streptomycin-colistin and colistin groups. Increased proportions of Parabacteroides goldsteinii, Bacteroides vulgatus, Bacteroides thetaiotaomicron, and Bacteroides uniforms were observed in the colistin group. The significant expansion of KCs was identified in the ampicillin-streptomycin-colistin and colistin groups. B.vulgatus levels were positively correlated with KC levels. More liver metastases were observed in the vancomycin group. An increased abundance of Parabacteroides distasonis and Proteus mirabilis and an obvious reduction of KCs were noted in the vancomycin group. P. mirabilis levels were negatively related to KC levels. The number of liver metastatic nodules was increased in the P. mirabilis group and decreased in the B. vulgatus group. The number of KCs decreased in the P. mirabilis group and increased in the B. vulgatus group. In vitro, as P. mirabilis or B. vulgatus doses increased, there was an opposite effect on KC proliferation in dose- and time-dependent manners. P. mirabilis induced CT26 cell migration by controlling KC proliferation, whereas B. vulgatus prevented this migration. @*Conclusions@#An increased abundance of P. mirabilis and decreased amount of B. vulgatus play key roles in CRC liver metastasis, which might be related to KC reductions in the liver.

Significance and Prospect of Tryptophan Metabolism in Treatment of Tumor Immune Checkpoint Inhibitors / 肿瘤防治研究

Although immune checkpoint inhibitors (ICIs) have great breakthrough in cancer treatment in recent years, most patients have not benefited from it on account of immune microenvironment. Studies have shown that tryptophan metabolism is not only involved in the formation of tumor immunosuppressive microenvironment but also plays an important role in the therapeutic application of ICIs. At present, inhibiting the kynurenine pathway of tryptophan metabolism is now in various stages of clinical trials, while the other two metabolic pathways, 5-HT and the indole pathway, also have aroused wide concern. This article reviews the latest developments in this field.

Effects of 5-hydroxytryptamine and receptors in tumorigenesis / 国际肿瘤学杂志

The role of 5-hydroxytryptamine (5-HT)has attracted more and more attention in the development of tumor. In addition to the synthesis of 5-HT by enterochromaffin cells,some tumor cells and im-mune cells can also secrete 5-HT,and there are different 5-HT receptors on the surface of these cells. 5-HT plays multiple biological effects through activating different receptors. 5-HT promotes the proliferation and inva-sion of tumor and induces the angiogenesis of tumor tissues through activating its corresponding receptors,such as 5-HT1D,and so on. Furthermore,various 5-HT receptors are expressed on the surface of immune cells. On the one hand,physiological doses of 5-HT promote the proliferation and secretion of immune cells. On the other hand,pathologic 5-HT has an effect on inducing the differentiation of immune cells in the direction of immuno-suppression,thus participating in the occurrence and development of tumor.

Effects of 5-hydroxytryptamine and receptors in tumorigenesis / 国际肿瘤学杂志

The role of 5-hydroxytryptamine (5-HT) has attracted more and more attention in the development of tumor. In addition to the synthesis of 5-HT by enterochromaffin cells, some tumor cells and immune cells can also secrete 5-HT, and there are different 5-HT receptors on the surface of these cells. 5-HT plays multiple biological effects through activating different receptors. 5-HT promotes the proliferation and invasion of tumor and induces the angiogenesis of tumor tissues through activating its corresponding receptors, such as 5-HT1D, and so on. Furthermore, various 5-HT receptors are expressed on the surface of immune cells. On the one hand, physiological doses of 5-HT promote the proliferation and secretion of immune cells. On the other hand, pathologic 5-HT has an effect on inducing the differentiation of immune cells in the direction of immunosuppression, thus participating in the occurrence and development of tumor.

Isolation of intraperitoneal free cancer cells from colorectal cancer by immunomagnetic beads / 中国基层医药

ObjectiveTo explore the value of immuomagnetic beads(IMB) technique for detection of intraperitoneal free cancer cells from colorectal cancer.MethodsPeritoneal lavage fluid was obtained from 80 patients with colorectal cancer during laparotomy.Peritoneal lavage cytology (PLC) and IMB were used to detect free cancer cells in peritoneal lavage fluid.10 patients with hysteromyoma during laparotomy were enrolled into the control group.ResultsThe positive rate of PLC was 8.8％ (7/80),the positive rate of IMB was 28.8％ (23/80).The positive case after useing PLC detect,IMB detect also was positive.The detected samples of control group were negative by these two methods.IMB was superior to PLC ( x2 =10.503,P =0.001 ).ConclusionIMB was more sensitive and specific than PLC,which could provide a effective method for finding intraperitoneal free cancer cells.

Biological changes of Kupffer cells in response to suppression of discoidin domain receptor 1 by in vivo delivery of small interfering RNA following acute hepatic injury / 中华创伤杂志

Objective To investigate the effect of small interfering RNA (siRNA) suppressing discoidin domain receptor 1 (DDR1) gene on biological behaviour of Kupffer cells (KC) in acute hepatic injury. Methods Male BALB/c mice were randomly divided into control, hepatic injury model, non-silencing siRNA and DDRlsiRNA groups. Hepatic injury model induced by intravenous injection of Con-canavalinA (ConA) 15 mg/kg, with or without hydrodynamic tail intravenous injection of naked siRNA (50 μg,2.0-2.5 mg/kg)/mouse or 1.5 ml normal saline. The expression of DDRI was assayed by West-ern blot and pro-inflammatory cytokine expression analyzed by ELISA. In the meantime, alanine amin-otransferase (ALT) and Kupffer cells'clearance to carbon granules was detected. Results The expres-sion of DDR1 obviously increasod at 6 h after hepatic injury, roached peak at 24 h and began to decrease at 48 h. Pretreatment with DDRisiRNA could obviously inhibit the expression of DDR1 and abrogate the high levels of ALT, expressions of TNF-α and IL-1β as well as phagocytosis of Kupffer cells. Conclu-sions Inhibition of discoidin domain receptor 1 by in vivo delivery of siRNA attenuates ConA-induced hepatic injury. Possible mechanism is that the inhibition of activity of KC inhibits the expression of pro-in-flammatory cytokines and thus alleviates hepatic injury.

Prophylactic and therapeutic effect of oxymatrine on D-galactosamine-induced rat liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the prophylactic and therapeutic effect of oxymatrine on experimental liver fibrosis and to reveal its mechanism.</p><p><b>METHODS</b>By establishing D-galactosamine-induced rat liver fibrosis model, we observed the effect of oxymatrine on serum and tissue biochemical indexes, content of liver hydroxyline, expression of TGF?1 mRNA and changes of tissue pathology.</p><p><b>RESULTS</b>There was a decline of liver hydroxyline and serum AST and ALT in oxymatrine group compared to those of the D-GalN group. The hydroxyline content in oxymatrine pretreatment group was (0.50 0.11)mug/mg compared with (0.99 0.14)mug/mg in D-GalN group (t=8.366, P<0.01). The content in oxymatrine treatment group was (0.44 0.04)mug/mg compared with 0.70 0.06 in D-GalN group (t=9.839, P<0.01). The SOD activity was (149.81 15.28) NU/mg in oxymatrine pretreatment group and (95.22 16.33) NU/mg in the model group (t=7.309, P<0.01); (157.68 19.54) NU/mg in the treatment group compared with (119.88 14.94) NU/mg in the model group (t=4.348, P<0.01). MDA in the pretreatment group was (2.06 0.17) nmol/mg, lower than (4.57 0.37) nmol/mg in the model group (t=17.529, P<0.01). In the treatment group, it was (1.76 0.24)nmol/mg, lower than (3.10 0.17) nmol/mg in the model group (t=12.697, P<0.01). TGF?1 mRNA reduced in the pretreatment and treatment groups as compared with that in the model group (0.21 0.01 vs 0.50 0.01, t=48.665, P<0.01; 0.18 0.02 vs 0.38 0.01, t=22.464, P<0.01). Electron microscopy showed that oxymatrine group had milder hepatocyte degeneration and less fibrosis accumulation than did the model group. Microscopy revealed wide septa expansion from the portal area to the central venous, piecemeal and confluent necrosis and pseudo-nodular formation in part of the lobular in the model group. While in oxymatrine group these lesions were much improved.</p><p><b>CONCLUSIONS</b>Oxymatrine shows prophylactic and therapeutic effect in D-galactosamine induced rat liver fibrosis. This is partly by protecting hepatocyte and suppressing fibrosis accumulation through anti-lipoperoxidation.</p>

Expression of desmin, GFAP and ?-SMA in human and RAT pancreatic stellate cells / 解放军医学杂志

Objective To investigate the dynamic changes in expression of cell markers desmin, glial fibrillary acidic protein (GFAP), and (-smooth muscle actin (?-SMA) in primary cultures of human and rat pancreatic stellate cells (PSCs). Methods PSCs were isolated from human as well as rat pancreas using Nycodenz discontinuous density gradient centrifugation following digestion with combination of collagenase IV, Pronase E and DNase I, and purified by centrifugal elution techniques. Freshly isolated cells were examined by laser scanning confocal microscopy for vitamin A autofluorescence, by immunostaining for desmin, GFAP, and ?-SMA. Expression of ?-SMA was as well measured by Western analysis. Procollagen ?1(Ⅰ) mRNA expression was analyzed by Northern analysis. Results The purity of rat PSCs obtained by centrifugal elution were above 95%. More than 85% of either freshly-isolated human or rat PSCs displayed positive vitamin A autofluorescence. Rat PSCs stained positively for desmin and GFAP and negatively for ?-SMA, whereas human PSCs were negative for either desmin, GFAP or ?-SMA. During the process of primary culture, rat PSCs were positive for ?-SMA at 3d and completely transformed from quiescent state to myofibroblast-like phenotypes at 7d, which negatively or scarcely expressed desmin and GFAP, but fully expressed the ?-SMA protein and procollagen ?1(Ⅰ) mRNA, similarly to the settings of human PSCs. Conclusions Human and rat PSCs could be successively isolated in above 95% purity by combining gradient centrifugation with following centrifugal elution techniques. The results show some species differences in desmin and GFAP expression between freshly-isolated human and rat PSCs. Both of which, however, acquire a myofibroblast-like phenotype largely expressing ?-SMA protein and procollagen ?1(Ⅰ) gene in culture.

Inhibitory effects of ?-lipoic acid on activation of NF-?B induced by high glucose in rat mesenteric cells / 解放军医学杂志

Objective To investigate the effects of ?-lipoic acid on the proliferation and activation of NF-?B induced by high glucose (HG) in rat mesenteric cells (MCs). Methods The rat mesenteric cells were cultured in the medium with normal glucose (5.6mmol/L, NG), high glucose (25mmol/L, HG), HG+100 ?mol/L ?-lipoic acid, or HG+200?mol/L ?-lipoic acid and HG+PDTC (a NF-?B inhibitor). Activation of nuclear factor-?B (NF-?B) of rat mesenteric cells were measured by electrophoretic mobility shift assay (EMSA). The cell proliferation was assessed by MTT. Results ?-lipoic acid (50~300?mol/L) can inhibit the proliferation of MCs. The NF-?B binding activity was 2.2 -fold higher in MCs exposed to HG compared to NG (P

Inhibitory effects of rosiglitazone on activation of NF-?B and expression of ICAM-1 induced by high glucose in rat mesangial cells / 中国糖尿病杂志

Objective To investigate the role of rosiglitazone in high glucose(HG)-induced intercellular adhesion molecule-1(ICAM-1) expression of rat mesangial cells.Methods The rat mesangial cells(MCs) were cultured in the medium with normal glucose(5.6 mmol/L,NG),high glucose(25 mmol/L,HG),HG+5 ?mol/L rosiglitazone,HG+20 ?mol/L rosiglitazone and HG+PDTC(?NF-?B inhibitor).ICAM-1 mRNA expression was measured by semi-quantitative RT-PCR assay.Activation of nuclear factor-?B(NF-?B) of rat mesangial cells was measured by electrophoretic mobility shift assay(EMSA).The levels of ICAM-1 in the supernatants were determined by enzyme-linked immunosorbant assay(ELISA.) Results RT-PCR results showed that high-glucose increased the ratio of PCR products of ICAM-1 over GAPDH to 2.9-fold,which was prevented by rosiglitazone(5 and 20 ?mol/L) pre-treatment.The NF-?B binding activity was 2.5-fold higher in MCs exposed to HG as compared with NG(P

Role of p38MAPK in production of pro-inflammatory cytokines in Kupffer cells from severe acute pancreatitis rats / 中国病理生理杂志

AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in the Kupffer cells (KCs) production of pro-inflammatory cytokines, tumor necrosis factor-?(TNF-?) and interleukin-1?(IL-1?), in severe acute pancreatitis (SAP) rats. METHODS: Sprague-Dwaley rats were randomized into three groups: ①sham operation rats, ②SAP rats, ③SAP rats given the p38 MAPK inhibitor CNI-1493(10 mg/kg, iv). The SAP model was induced by the bili-pancreatic duct infusion with 5% sterile soduim taurocholate solution. Rats from each group were killed at 12 h after sham operation or SAP and Kupffer cells (KCs) were isolated. The mRNA expressions of TNF-? and IL-1? (by quantitative real-time RT-PCR) and p38 MAPK activity (by Western blot analysis) in KCs were examined. The levels of TNF-? and IL-1? in plasma were determined by ELISA. RESULTS: There was a significant acvitation of p38 MAPK in KCs harvested from SAP rats than those from sham operation rats. SAP also promoted the mRNA expressions of TNF-? and IL-1? in KCs and the plasma levels of TNF-? and IL-1?. These events were significantly inhibited by treatment with CNI-1493.CONCLUSIONS: p38 MAPK activation is one important aspect of the signaling events that may mediate the KCs production of pro-inflammatory cytokines, TNF-? and IL-1?, in SAP rats. The inhibition of the p38 MAPK may be a potential target in the prevention and treatment of SAP.

Effects of tempol on p38MAPK activation of gastric mucosa ulceration in rat after water-immersion restraint stress / 解放军医学杂志

Objective To investigate the temporal activation of p38MAPK signaling transduction cascade,and the relationship between the activation and reactive oxygen species(ROS)in a well-defined experimental model of water-immersion restraint(WIR)stress-induced gastric mucosa ulceration.Methods Male Sprague-Dawley rats were randomly divided into control group,WIR group and tempol treated group.Animals were restrained and immersed in water bath to induce gastric mucosal lesions,with or without pretreatment with the free radical scavenger,tempol,and the WIR group animals were killed at different time points after WIR stress.Tempol treated animals underwent the same protocol followed 30min or 360min of stress,then the gastric mucosa was harvested,and the activity of gastric mucosal p38 was analyzed by Western blot.Malondialdehyde(MDA)activity and pro-inflammatory cytokine expression were detected.Results A rapid activation of p38MAPK in gastric mucosa occurred as early as 15 min after stress,and this activation was maximal after 90min of stress and still persisted until 360min after stress.Pretreatment with tempol prevented stress-induced p38MAPK activation at 30 min and 360min time points(0.77?0.24 and 0.58?0.12,respectively)compared with that in model groups(1.22?0.16 and 1.73?0.09,respectively,P