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OBJECTIVE: To observe the effects of ginsenoside (Gs) and berberine (Ber), two kinds of active components of traditional Chinese herbal medicine, on transforming growth factor-beta1 (TGF-beta1) and prostaglandin E2 (PGE2) in PG cells. METHODS: Co-culture system of human lung carcinoma cell line PG and human T lymphocyte cell line Jurkat was established. PG cells were treated with Gs (100 microg/ml) and Ber (10 mug/ml) for twenty-four hours, and then cocultured with Jurkat cells. After 24-hour coculture, the state of Jurkat cells was observed with inverted microscope. The viable count of Jurkat cells was detected by trypan blue staining after 6- and 24-hour coculture, and the apoptosis of Jurkat cells was evaluated by flow cytometry. PG cells were treated with 100, 50, 25 microg/ml Gs and 10, 5, 2.5 microg/ml Ber respectively, and the content of TGF-beta1 and PGE(2) in PG cells was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) method. RESULTS: After coculture with PG cells treated with Gs and Ber, the number of Jurkat cells was less than blank control group, and the apoptosis rates of Jurkat cells in Gs- and Ber-treated groups were higher than blank control group. Gs and Ber could promote the secretion of TGF-beta1 in PG cells, but could not change the level of PGE(2). CONCLUSION: Gs and Ber can promote the growth inhibition and apoptosis of Jurkat cells induced by PG cells, which may be related to the up-regulation of Gs and Ber on TGF-beta1 secretion in PG cells.
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AIM: To study the effect of berberine(Ber) on invasion and migration of PG cells from a high metastatic human giant lung carcinoma cell line and to explore its mechanism. METHODS: Agarose drop method was used to detect PG migration; transwell cabin with FN in lower chamber was adopted to detect PG chemotaxis. PG adhesion to FN and martrigel was detected by MTT; PG invasive ability was determined by transwell cabin covered with martrigel. Expression of MMP2/TIMP2 protein and mRNA were detected by quantitative immunocytochemical method and RT - PCR respectively. RESULTS: After PG was treated by Ber( 10 mg/L) for 24 h: 1 ) migration distance of Ber- treated PG cells was markedly shorter than that of control cells (P ＜0. 01 ) and the number of passed membrane cells towards FN was much fewer than that of control cells ( P ＜ 0. 01 ); 2) PG adhesion to FN and martrigel was inhibited remarkably by Ber compared with control PG; 3) the migration of PG cells through the martrigel -coated transwell was significantly inhibited by the addition of Ber; 4) MMP2 expression was reduced significantly(P ＜0. 01 ), while the TIMP2 expression showed up - regulating tendency, but had no differences compared with control group (P ＞ 0. 05). The MMP2/TIMP2 ratio was decreased; 5 )the MMP2 mRNA/TIMP2 mRNA ratio was decreased by Ber. CONCLUSION: Inhibition of cell migration, adhesion to ECM and invasion into ECM of tumor cells and regulation of homeostasis between MMPs and TIMPs to maintain ECM integrity may be the basic mechanism of inhibitive effect of Ber on invasion and metastasis of tumors.
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AIM: To investigate the effect of phellodendron and three kinds of its main components, which have asuppressive effect on the immune system, on the membrane fluidity of normal murine splenocytes. METHODS: The fluidity ofmembrane lipid regions of splenocytes was determined by the fluorescence polarization technique using 1, 6 - diphenyl - 1, 3, 5- heatriene (DPH) as a fluorescence probe. RESULTS: The results showed that the water extract of phellodendron and one of itsmain components (palmatine) increased the cell membrane fluidity in the inactive state, but the other two components, berberineand jatrorrhizine, decreased the cell membrane fluidity. After activated by ConA, all of them can decrease the cell membrane flu-idity. CONCLUSION: These results suggest that their immunosuppressive function might be due to decreasing the cell membranefluidity.
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AIM: To investigate the anti - tumor effect of Weimaining (WMN) on a murine Lewis lung carcinoma (3LL) and the influence on the cell cycle. METHODS: The inhibitory rate of WMN in 3LL growth was detected by replicating the model of 3LL. The effect of the drug on 3LL cell cycle and the influence of the drug on the expression of cy clin D1 protein were investigated by flow cytometry and immunohistochemical staining. RESULTS: The results showed that the inhibitory rate of drug in 3LL is 19. 14%, 33.59%, 40. 63% and 51.56% respectively at dosage ranging from 100,cells in G0 -G1 phase and decreases the expression of cyclin D1 protein. CONCLUSION: WMN inhibits the growth of 3LL cells in vivo by decreasing the expression of cyclin D1, blocking the cells in G0 - G1 phase and preventing the cells transition from G1 to S phase while DNA is replicated.
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AIM:To investigate the effect of berberine on IL-1 or tumour necrosis factor (TNF) induced polymorphonuclear leucocyte(PMN)-endothelium adhesion and adhesion molecules.METHODS:Based on the model of human umbilical vein endothelial cell (HUVEC), this study adopted Rose Bengal Stain, cell ELISA, immunocyto-chemical techniques to investigate the effect of berberine on PMN-endothelium adhesion and the expression of cell adhesion molecules (CAMs).RESULTS:Berberine inhibited IL-1, TNF-induced HUVEC adhesion for PMN when pretreated HUVEC and antagonised IL-1, TNF-induced upregulation of ICAM-1 on HUVEC. Meanwhile, TNF-stimulated PMN adhesion for HUVEC and CD18 upexpression on PMN was diminished in the presence of berberine.CONCLUSION: Inhibite PMN-endothelium adhesion by downregulating the CAMs expression to inhibite PMN migration across endothelium is one of the mechanisms of antiinflammation of berberine.
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AIM: To study the effect of berberine on the proliferation of human pulmonary carcinoma cells (PG cells). METHODS: The proliferation of PG cells was determined by using MTT assay. Cell cycle and apoptosis of PG cells were determined by using flow cytometry. Confocal scanning imaging system was used to assay the ROS-releasing level of PG cells. RESULTS: Berberine was shown to inhibit proliferation of PG cells directly and in a concentration-dependent manner (P
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AIM: To investigate the effect of Qingjiening(QJN) on cytokines in sheep red blood cell(SRBC)-immunized mice. METHODS: After immunization of mice with SRBC, the effect of QJN o n IL-1、IFN-?、IL-2 in mice was observed, the IFN-? level was measured by macrophage NO - 2-release assay, the IL-1 level was measured by thymocyte a ssay, the IL-2 level was measured by mitogen activated lymphocytoblast assay. RESULTS: QJN can significantly inhibit mice to secrete IL-1、IFN- ? and IL-2. CONCLUSION: The immunosuppressive activity of QJN may be associate d with inhibition of immunocyte to secret IL-1、IFN-? and IL-2.
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AIM: To investigate the mechanism of berberine’s inhibiting growth and metastasis of tumor by observing the effects of berberine on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were incubated with berberine. MTT assay and quantitative immunocytochemistry were used to detect cell proliferation and the expression of proliferation cell nuclear antigen (PCNA). Morphologic changes of apoptosis were observed by fluorescent staining, and Rhodamin123 was used to determinate mitochondrial membrane potential under the laser confocal microscopy. RESULTS: HUVEC proliferation was inhibited by co-incubating with berberine (20 mg/L) for 24 h and berberine (10 mg/L, 20 mg/L) for 48 h (P
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AIM: To observe the effects of some component of Chinese herbs for external use on proliferation of human umbilical vein endothelial cells (HUVEC) and investigate the mechanism of promoting tissue repair. METHODS: The method of MTT was used to examine the effects of Rg1, Rh1, perlolyrine, cinnamyl aldehyde, muscone, astragalus polysaccharin (APS), velver antler polypeptide (VAP) and soluble extract of boswellia carterii birdw (BCB) on proliferation of HUVEC. RESULTS: APS did not promote proliferation of HUVEC at 9.75 mg/L-2.5 g/L; Rh1 promoted proliferation of HUVEC at 1.94 mg/L-0.5 g/L (P
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AIM: To investigate the anti-metastasis effect of weimaining, extracted from fragopyrum cymosum meissn, a Chinese medicine, on murine Lewis lung carcinoma (3LL). METHODS: The anti-metastasis effect of weimaining in vivo was detected in the grafting lung metastasis model of murine Lewis lung carcinoma. The effects of the drug on the expression of CD34 and E-cadherin were investigated by immunohistochemical staining and RT-PCR. RESULTS: Weimaining effectively inhibited the lung metastasis of 3LL at a concentration of 250 mg?kg -1?d -1, significantly suppressed the expression of CD34 and increased the expression levels of E-cadherin protein and mRNA in 3LL cells. CONCLUSIONS: Weimaining inhibits the metastasis of murine Lewis lung carcinoma (3LL) in vivo via increasing the expression of E-cadherin and decreasing microvessel density of tumor tissue.
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Objective:To study the cytokine mechanism of Jiawei Xuanfei Toujie Ji(JXT)therapy to influenza infected mouse.Methods:Mice were inoculated intranasally with mouse lung-adapted influenza virus strain(FM1),and were treated with JXT. At 5th days after postinoculation were sacrificed, and IL-2,TNF-?,IL-6,IFN-? in their sera were measured with ELISA kits, their lungs sections were stained with hematoxylin and eosin.Results:Compared with model group ,the levels of IL-2 and IFN-? in JXT group were dramatically higher,and the levels of TNF-? and IL-6 were lower,and pathological effects of influenza pneumonia showed less lung injury.Conclusion:Regulative effects on cytokine may be one of the factors that JXT decrease the pathogenesis of influenza-induced acute lung injury.
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AIM:To study the effect of Yiqi Huoxue Jiedu Fang(YHJ),composed of ginsenoside,penex notogingseng and berberin,on tumor growth and metastasis and to explore its mechanism.METHODS:Murine Lewis lung carcinoma transplant model was established and mice were treated with YHJ by intraperitoneal injection.After 10 days,the inhibitory rate of tumor,pathology of tumor and PCNA of tumor cells were detected.After 20 days,numbers of metastatic foci on lung surface and microvessel density(MVD)were determined.Expression of VEGF in tumor and serum were also analyzed by immunohistochemical test and ELISA,respectively.RESULTS:YHJ reduced the weight of tumor and the amount of metastatic foci.The inhibitory rates of tumor at high and low dose of YHJ(24 mg?kg-1?d-1,12 mg?kg-1?d-1)were 48.29% and 37.26%,and the number of metastatic foci was 1.67 and 3.50,while control was 6.44.Furthermore,PCNA of tumor cells,MVD of tumor and VEGF expression in serum and tumor were decreased in YHJ treatment goup as compared with control.CONCLUSION:YHJ remarkably inhibits Lewis lung carcinoma growth and metastasis in mice.Its mechanism may be related to inhibition of angiogenesis.
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Objective:To test the effect of Ginsenoside Rg1 and Rh1 on the expression of CD25,HLA-DR,CD44,ICAM-1,CD11c and E-selectin on the surface of dendritic cell(DC).Methods:Mature DCs were treated with different doses of Rg1 and Rh1. Certain time later, the expression of adhesion molecular was detected with immunohistochemical method and the results were recorded with imaging data.Results:Except for E-selectin, both of Rg1 and Rh1 could increase the expression of HLA-DR,CD25,CD11c,CD44 and ICAM-1 on DCs.Conclusion:Expression increase of surface molecular, including in two signals required by T cell activation, stimulated by Rg1 and Rh1, is propitious to the formation of DC- T cell cluster that would enhance the antigen-presenting function of DC.
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Objective:Based on the model of human umbilical vein endothelial cell(HUVEC) treated by tumor necrosis factor-alfa(TNF),We investigate the effects of muscone on polymophonulear leukocytes(PMN)-HUVEC adhesion and its adhesion molecules(CAMs) expression.Methods:Confocal system was used for identifying cultured HUVEC,MTT assay for its activity,Rose Bengal Staining for PMN-HUVEC adhesion,and fluorescent-immunocytochemistry techniques for CAMs expression.Results:After HUVEC treated by TNF,the adhesion between PMN and HUVEC increased dramatically(P