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Objective:To investigate the effects of 30% ethanol elution fraction of Artemisia absinthium extract with macroporous resin (AAEM-30%) on the dendritic cell (DC) and immunity of mice. Methods:AAEM-30% was obtained from the alcoholic extracts of A. absinthium by AB-8 macroporous resin, and its polysaccharide, flavonoid, and terpenoid contents were determined. The expressions of AAEM-30% on DC surface molecular cluster of differentiation (CD) 40, CD80 and CD86 were detected in vitro by flow cytometry, and the expressions of DC cytokines IL-6 and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). The effect of AAEM-30% on the immune function of ICR mice was measured in vivo with different doses (50 and 100 mg/kg) and different administration methods (subcutaneous injection, intraperitoneal injection, and gavage). Results:The contents of polysaccharides, flavonoids, and terpenoids in AAEM-30% were 24.30%, 22.50% and 28.19%, respectively. AAEM-30% significantly enhanced the expression of CD40, and CD86 and the secretion of IL-6 and TNF-α (all P<0.001). Compared with the control group, no statistically significant differences were found in the body mass of mice compared with the three administration methods (all P>0.05). The thymus index in the 50 and 100 mg/kg AAEM-30% intraperitoneal injection groups and the spleen index in the 50 mg/kg AAEM-30% gavage group were increased (all P<0.05). CD19 + cells increased in the 100 mg/kg AAEM-30% intraperitoneal injection group ( P<0.01) and in the 50 mg/kg AAEM-30% gavage group ( P<0.05). The CD11b + and CD11c + counts increased in the 100 mg/kg AAEM-30% gavage group ( P<0.05). The number of CD4 + and CD8 + T lymphocytes was increased by both gavage and intraperitoneal administration (all P<0.05). Conclusions:AAEM-30% can promote the maturation of DC and enhanced the immunity of mice without obvious side effects.
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Objective To investigate the inhibitory effect of cryptotanshinone (CPT) on human breast cancer cell MCF7 and its mechanism. Methods The survival rate of MCF7 cells was measured by MTT assay. Cell apoptosis was detected by Annexin V/PI assay and Hoechst 33258 fluorescence staining assay. Cell cycle and reactive oxygen species were detected by flow cytometry. Cell migration and invasion were detected by cell scratch test and Transwell chamber test. The surface molecules CD44 and CD24 were detected by flow cytometry and microsphere culture. The expression of cell-associated proteins was detected by Western blot. Results CPT inhibited the proliferation of MCF7 cells in a dose-dependent manner, and the 24 h IC50 value was 19.24 μmol/L. Compared with the untreated group, the CPT-treated group showed cell cycle arrested in the S phase, and apoptosis was induced. The results of the cell scratch and Transwell chamber tests showed that CPT significantly inhibited the migration and invasion of MCF7 cells. Furthermore, CPT reduced the CD24-/LowCD44+ cell population in MCF7 cell-derived microspheres. Western blot results showed that CPT could up-regulate the expression of Bax protein, down-regulate the expression of BCL-2, PI3K-p85, Akt, N-cadherin, Twist1, Sox2, Oct4, and Nanog protein, effectively inhibit the phosphorylation of ER-α, and decrease the expression of ABCG2. Conclusion CPT can inhibit the proliferation of MCF7 cells by inhibiting the migration and invasion of MCF7 cells, decreasing the number of CD24-/lowCD44+ cells and affecting the expression of tumor stem cell-related proteins.
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Objective:To investigate the immunoregulatory effects of Glycyrrhiza uralensis ethanol extract(GUEE) on the maturation of dendritic cells (DCs) and the adjuvant effect of GUEE on OVA in na?ve BALB/c mice and an ovalbumin (OVA)-induced asthma mouse model. Methods:GUEE was prepared, and the effects of different concentrations of GUEE on the maturation of DCs and the secreted cytokines as well as the effects of GUEE on bacterial lipopolysaccharide (LPS)-induced DC maturation were examined in vitro. The effect of GUEE on the morphology of mouse bone marrow derived DCs was observed using microscopy. Molecular expression levels on the surface of DCs were detected using flow cytometry. The levels of interleukin-1β (IL-1β), IL-6, IL-12, and tumor necrosis factor-α (TNF-α) in the supernatant of DCs cultures were measured by enzyme-linked immunosorbent assay (ELISA). The maturation status of DCs was detected by flow cytometry by injecting different concentrations of GUEE into the paws of mice and isolating the draining lymph nodes 24 h later. The naive BALB/c mice were co-immunized with OVA, and the changes in regulatory T cells (Treg) were detected by flow cytometry. An OVA-protein-induced mouse asthma model was established to investigate whether GUEE as a tolerogenic adjuvant has an antigen-specific therapeutic effect on asthmatic mice. Pulmonary pathological changes were analyzed by hematoxylin-eosin staining (HE) and PAS staining. OVA-specific antibodies in serum and the frequencies of Tregs, CD4 + IFN-γ + and CD4 + IL-4 + T cells in the spleen were detected by ELISA and flow cytometry, respectively. Results:GUEE suppressed DCs maturation induced by LPS both in vitro and in vivo (all P<0.05), and reduced proinflammatory cytokine production, including IL-1β, IL-6, IL-12 and TNF-α in the absence or presence of LPS (all P<0.05). Moreover, co-immunization with OVA and GUEE increased the amount of Tregs in na?ve BALB/c mice ( P<0.05). In OVA-induced asthmatic mice, OVA and GUEE co-immunization and GUEE alone treatment substantially ameliorated the inflammation of lung tissues, decreased the levels of IgG 1 and the amount of CD4 + IL-4 + T cells, and increased the amount of Tregs (all P<0.05). Conclusions:GUEE alone or as the tolerogenic adjuvant can ameliorate allergic diseases through inhibition of DC maturation and type 2 helper T cell responses and induction of Tregs.
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Objective:To investigate the effect of silica gel column separation component of Artemisia asiatica (AEM-SC) on the maturation and immune function of mouse dendritic cells (DCs). Methods:Artemisia asiatica components were prepared by macroporous resin eluted with 70% ethanol, and then isolated by silica gel column to obtain AEM-SC. The contents of polysaccharides, flavonoids and triterpenes were quantified. Flow cytometry was used to detect the expression level of DCs surface molecules and antigen phagocytosis ability and to stimulate the proliferation of allogeneic T cells. ELISA method was used to detect the effect of DCs on cytokine secretion. Results:The contents of polysaccharides, flavonoids and triterpenes in AEM-SC were 10.12%, 5.7% and 3.62%, respectively. Functional tests showed that AEM-SC significantly reduced the expression levels of LPS-induced DCs surface molecules CD40, CD86 and MHC-II, reduced the expression levels of inflammatory cytokines IL-12p40, TNF-α and IL-6 (all P<0.05), improve the ability of phygocytosis ( P<0.01), and reduce the ability of DCs to stimulate the proliferation of CD4 +T and CD8 +T lymphocytes in the spleen of mice (all P<0.001). In the inflammatory mouse model experiment, AEM-SC significantly reduced the expression levels of DCs surface molecules CD40, CD86, CD80 (all P<0.001), and the expression levels of inflammatory cytokines TNF-α, IL-12p40 in serum (all P<0.01). Conclusions:AEM-SC can inhibit the maturation of DCs-induced LPS both in vitro and in vivo, indicating that AEM-SC has the immunosuppressive effect.
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Objective To investigate the effects of Artemisia absinthium L. ( A. absinthium L. ) extracts on the maturation and function of dendritic cells (DCs). Methods A. absinthium water (AAW) and ethanol ( AAE) extracts were prepared. DCs were separated into several groups and treated with differ-ent concentrations of AAW (containing 5, 50 and 150μg/ml of polysaccharide) and AAE (containing 0. 6, 3 and 6 μg/ml of flavonoid) , respectively. Cell viability, antigen phagocytosis and maturation of DCs were detected by flow cytometry. Levels of cytokines were analyzed by ELISA. Western blot assay was performed to analyze the activation of key molecules in NF-κB, mitogen-activated protein kinase ( MAPK) and janus kinase/signal transducer and activator of transcription ( JAK/STAT) signaling pathways. Results AAW promoted the maturation of DCs, significantly decreased antigen phagocytosis and increased cytokine produc-tion (IL-12p40 and TNF-α). AAE significantly enhanced the expression of co-stimulatory molecules on the surface of DCs and decreased antigen phagocytosis, but had no significant effect on cytokine production. Mo-reover, AAE significantly inhibited the LPS-induced expression of TNF-α, IL-12p40 and IL-6. Further anal-ysis revealed that AAW and AAE could activate the phosphorylation of p38, extra-cellular signal regulated kinase (ERK), IKKα/β, NF-κBp65 and JAK2. Besides, AAE could activate the phosphorylation of c-Jun N-terminal kinase ( JNK ) and inhibit the LPS-induced phosphorylation of inhibitor of NF-κB ( IκB-α) , IKKα/β, NF-κBp65, p38, ERK and JAK2. Conclusion AAW could enhance immunity and AAE could inhibit inflammation.
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Objective To investigate the effects of water extracts of Lepidium meyenii walp (LMWE) collected from two different places in Xinjiang on the maturation and function of dendritic cells (DCs) and to evaluate their roles in immunoregulation. Methods Water extracts of LMWE growing in Tashikuergan County (Ta xian) and A La gou of Xinjiang were prepared and named as LMWE-T and LMWE-A,respectively. Bone marrow-derived DCs and splenocytes isolated from C57BL/6 mice were treated with different concentrations of polysaccharide extracts from LMWE-T/A. Effects of LMWE-T/A on the per-centage and apoptosis of DC,expression of co-stimulatory molecules and secretion of cytokines were detected by flow cytometry and ELISA. MTT assay was used to analyze the proliferation of splenocytes. Changes in the functions of DC were evaluated by mixed lymphocyte reaction(MLR). Results LMWE-T/A had no in-fluence on the percentage and viability of DC. Expression of CD40 and CD86,and secretion of TNF-α,IL-12p40 and IFN-γ were significantly increased by LMWE-T/A treatment in a dose-dependent manner. LMWE-T/A treatment enhanced the functions of DCs and also dose-dependently promoted the proliferation of splenocytes. LMWE-A was more effective than LMWE-T in promoting CD86 expression,IFN-γ secretion and splenocyte proliferation. Pretreatment with TAK-242,an inhibitor of Toll-like receptor 4(TLR4),could sig-nificantly inhibit the regulatory effects of LMWE-T/A on CD40 expression and the secretion of TNF-α and IL-12p40. Conclusion This study shows that LMWE could promote the maturation of DC through TLR4 signaling pathway,enhance the functions of DC without side effects on DC survival,and increase the prolif-eration of splenocytes,indicating that LMWE has a potential immunopotentiating effect. LMWE-A has better effects than LMWE-T on immune enhancement.
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Glypican-3 ( GPC3 ) plays very important role in the regulation of cell growth and differentiation in hepatocellular car-cinoma ( HCC ) . GPC3 is closely related to the occurrence and development of HCC. A dramatic elevation of GPC3 expression has been reported in a large proportion of HCC, which suggests that GPC3 is remarkably sensitive and specific to the diagnosis of HCC. GPC3 is a potential therapeutic target of HCC. This paper reviews the structure and function of GPC3, the progress of im-munotherapy based on GPC3 of HCC, and discusses the prospect of therapeutic target of liver cancer in the future.
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Squamous cell carcinoma is the second most common non -melanoma skin cancer and its morbidity is increasing in recent years .Because of the complicated pathogenesis and non -specific clinical mani-festation of squamous cell carcinoma ,misdiagnosis occurs frequently .Although squamous cell carcinoma usually grows very slowly ,the invasion and metastasis of special types occurs early .With the deep knowledge of squamous cell carcinoma,people get more and more research results in diagnosis ,treatment and prevention .This paper de-scribes the epidemiology ,etiology ,clinical manifestation ,pathology ,clinical staging ,treatment and prognosis .
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Aim To investigate the apoptosis of human gastric carcinoma AGS cells induced by cecropinXJ. Methods Human gastric carcinoma AGS cells and human normal epithelial cells GES-1 were co-cultured with different concentrations of cecropinXJ ranging from 0. 01 to 1 000 mg·L-1 for 24 h. MTT assay was used to observe the effects of cecropinXJ on the proliferation of AGS cells and GES-1 cells. The ultrastructural changes of the AGS cells were observed by transmission electron microscopy. Hoechst staining was used to de-tect cell apoptosis. The changes of intracellular reac-tive oxygen species ( ROS) and mitochondrial potential were analysed by flow cytometery. The expression of Bax, Bcl-2, caspase-3 and cytochrome C in mRNA level was investigated by qRT-PCR. Western blot was used to determine the protein expression of Bax, Bcl-2, caspase-3 and cytochrome C. Results CecropinXJ significantly suppressed the proliferation of AGS cells in vitro (P<0. 05) in a dose-dependent manner, IC50 =61. 19 mg·L-1 , but had no inhibitive effects on the proliferation of GES-1 cells. After treatment for 24 h, cecropinXJ induced AGS cells nuclear condensation, and increased ROS production, disrupted mitochondri-al integrity. The results of qRT-PCR and Western blot demonstrated cecropinXJ could up-regulate the expres-sion of Bax and down-regulate the expression of Bcl-2 , promote the release of cytochrome C and activate caspase-3. Meanwhile, cecropinXJ promoted caspase-3 activity in a dose-dependent manner, and cell death ratio of AGS cells induced by cecropinXJ was signifi-cantly reduced by caspase-3 and caspase-9 specific in-hibitors treatment. Conclusion CecropinXJ can in-duce apoptosis of AGS cells by downregulating Bcl-2 , upregulating Bax and activating caspase-3 , which may be one of its anti-tumor mechanisms.
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Objective:Antimicrobial peptides ( AMPs ) are peptides generated in the biological defense system against exogenous pathogens ,which is all important constituent of the non-specific immune system.The immune system can react and sometimes play an important role in tumor control both in animal models and in humans.In this study,CecropinXJ were used in the BALB/c Eca109 cells-bearing mice model.To investigate whether CecropinXJ exert anti-tumor efficiency through regulating immune response when treated with tumor-bearing mice.Methods:The expression of cytokines and DC surface molecules were detected by immunohisto-chemistry and Flow cytometry.Results:The experiment of mice showed that CecropinXJ could significantly inhibit the proliferation of transplanted tumors in dose-dependent manner.There were significant difference between control group and 10mg/kg CecropinXJ group on the volume and weight of tumor ( P0.05).In CecropinXJ treatment group,TNF-αlevel in serum was significantly higher than the control group ( P<0.01 ).Conclusion: CecropinXJ could significantly inhibit tumor growth in Eca109 cells-bearing mice,and might not be affected anti-tumor efficiency through regulating immune response.