Objective To study the effect of Asparagus officinalis polysaccharide on the number and activity of erythrocyte complement receptor 1 in S180 mice.Methods Red blood cells from mice venous blood were labeled by rat ant...Objective To study the effect of Asparagus officinalis polysaccharide on the number and activity of erythrocyte complement receptor 1 in S180 mice.Methods Red blood cells from mice venous blood were labeled by rat anti-mouse CD35 monoclonal antibody and FITC-conjugated goat anti-mouse antibody.Using flow cytometry,we determined the number of ECR1.Using microscope,we studied the adherence between erythrocyte immunity and C3b receptor or tumor-cell by RBC-C3bRR and DTER.Results Comparing the mean value of the number of CR1 on each RBC of high and middle groups with control groups,the mean value of the number of CR1,RBC-C3bRR and DTER of Asparagus officinalis polysaccharide groups are increased significantly.Conclusions Asparagus officinalis polysaccharide can improve the erythrocyte function of S180 mice,which may be one of its most important antitumor mechanisms.展开更多
Invasive fungal infections are life-threatening,and neutrophils are vital cells of the innate immune system that defend against them.The role of LTA4H-LTB_(4)-BLT1 axis in regulation of neutrophil responses to fungal ...Invasive fungal infections are life-threatening,and neutrophils are vital cells of the innate immune system that defend against them.The role of LTA4H-LTB_(4)-BLT1 axis in regulation of neutrophil responses to fungal infection remains poorly understood.Here,we demonstrated that the LTA4H-LTB_(4)-BLT1 axis protects the host against Candida albicans and Aspergillus fumigatus,but not Cryptococcus neoformans infection,by regulating the antifungal activity of neutrophils.Our results show that deleting Lta4h or Blt1 substantially impairs the fungal-specific phagocytic capacity of neutrophils.Moreover,defective activation of the spleen tyrosine kinase(Syk)and extracellular signal-related kinase(ERK1/2)pathways in neutrophils accompanies this impairment.Mechanistically,BLT1 regulates CR3-mediated,β-1,3-glucan-induced neutrophil phagocytosis,while a physical interaction with CR3 with slight influence on its dynamics is observed.Our findings thus demonstrate that the LTA4H-LTB_(4)-BLT1 axis is essential for the phagocytic function of neutrophils in host antifungal immune response against Candida albicans and Aspergillus fumigatus.展开更多
Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein Barr virus The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen activated pro...Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein Barr virus The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen activated protein kinases (MAPKs, including ERK, JNK, and p38MAPK), and to highlight the molecular mechanism of CD 4 + cell deletion in AIDS Methods HOS cells (HOS CR2) and HOS CD4 cells (HOS CD4CR2) stably expressing CR2 were established and then identified by FACS and Western blotting Activation and blocking tests of MAPKs were assessed by Western blot Cell proliferation was determined using Cell Titer 96 Aqueous One Solution Reagent Results FACS results showed that the positive rates of HOS CR2 and HOS CD4CR2 cells were greater than 96%, and Western blot showed that the CR2 expression levels on HOS CR2 and HOS CD4CR2 cells were high Activation and blocking tests of MAPKs (ERK, JNK, and p38MAPK) were carried out in HOS CR2, HOS CD4, and HOS CD4CR2 cells The activation of MAPKs in HOS CR2 cells stimulated with PMA (100 ng/ml) and NHS (10%) was identical The activation of MAPKs increased at 5 minutes, reached a peak at 10 minutes, and decreased to baseline within 30 minutes, all in a time dependent manner; the activation of MAPKs was blocked by anti CR2 McAb, PD98059 (inhibitor of ERK), and Wortmanin (inhibitor of PI 3K), respectively In HOS CD4 cells, MAPKs were activated by HIV gp160 In HOS CD4CR2 cells, MAPK activation was induced by HIV gp160, 10% NHS, and HIV gp160+10%NHS; phosphorylation of p38MAPK was dramatically induced by HIV gp160+NHS, and lasted for 1 hour The cell proliferation results showed that HIV gp160 inhibited the proliferation of HOS CD4 and HOS CD4CR2 cells ( P <0 01) and that NHS enhanced the effect of HIV gp160 ( P <0 01) Conclusions The activation of MAPKs is independently mediated by CR2 and that anti CR2 McAb, PD98059, and Wortmanin block the activation of MAPKs, respectively The results of the signal transduction and cell proliferation assays of HOS CD4CR2 cells show that CR2 plays a role in the pathogenesis of HIV infection, especially in the inhibition of CD 4 + cell proliferation展开更多
AIM: To investigate the molecular mechanisms underlying the reversal effect of emodin on platinum resistance in hepatocellular carcinoma. METHODS: After the addition of 10 μmol/L emodin to HepG2/oxaliplatin (OXA) cel...AIM: To investigate the molecular mechanisms underlying the reversal effect of emodin on platinum resistance in hepatocellular carcinoma. METHODS: After the addition of 10 μmol/L emodin to HepG2/oxaliplatin (OXA) cells, the inhibition rate (IR), 50% inhibitory concentration (IC 50 ) and reversal index (IC 50 in experimental group/IC 50 in control group) were calculated. For HepG2, HepG2/OXA, HepG2/OXA/T, each cell line was divided into a control group, OXA group, OXA + fibroblast growth factor 7 (FGF7) group and OXA + emodin group, and the final concentrations of FGF7, emodin and OXA in each group were 5 ng/mL, 10 μg/mL and 10 μmol/L, respectively. Single-cell gel electrophoresis was conducted to detect DNA damage, and the fibroblast growth factor receptor 2 (FGFR2), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) and excision repair cross-complementing gene 1 (ERCC1) protein expression levels in each group were examined by Western blotting. RESULTS: Compared with the IC50 of 120.78 μmol/L in HepG2/OXA cells, the IC 50 decreased to 39.65 μmol/L after treatment with 10 μmol/L emodin; thus, the reversal index was 3.05. Compared with the control group, the tail length and Olive tail length in the OXA group, OXA + FGF7 group and OXA + emodin group were significantly increased, and the differences were statistically significant (P < 0.01). The tail length and Olive tail length were lower in the OXA + FGF7 group than in the OXA group, and this difference was also statistically significant. Compared with the OXA + FGF7 group, the tail extent, the Olive tail moment and the percentage of tail DNA were significantly increased in the OXA + emodin group, and these differences were statistically significant (P < 0.01). In comparison with its parental cell line HepG2, the HepG2/OXA cells demonstrated significantly increased FGFR2, p-ERK1/2 and ERCC1 expression levels, whereas the expression of all three molecules was significantly inhibited in HepG2/ OXA/T cells, in which FGFR2 was silenced by FGFR2 shRNA. In the examined HepG2 cells, the FGFR2, p-ERK1/2 and ERCC1 expression levels demonstrated increasing trends in the OXA group and OXA + FGF7 group. Compared with the OXA group and OXA + FGF7 group, the FGFR2, p-ERK1/2, and ERCC1 expression levels were significantly lower in the OXA + emodin group, and these differences were statistically significant. In the HepG2/OXA/T cell line that was transfected with FGFR2 shRNA, the FGFR2, p-ERK1/2 and ERCC1 expression levels were significantly inhibited, but there were no significant differences in these expression levels among the OXA, OXA + FGF7 and OXA + emodin groups. CONCLUSION: Emodin markedly reversed OXA resistance by enhancing OXA DNA damage in HepG2/OXA cells, and the molecular mechanism was related to the inhibitory effect on ERCC1 expression being mediated by the FGFR2/ERK1/2 signaling pathway.展开更多
文摘Objective To study the effect of Asparagus officinalis polysaccharide on the number and activity of erythrocyte complement receptor 1 in S180 mice.Methods Red blood cells from mice venous blood were labeled by rat anti-mouse CD35 monoclonal antibody and FITC-conjugated goat anti-mouse antibody.Using flow cytometry,we determined the number of ECR1.Using microscope,we studied the adherence between erythrocyte immunity and C3b receptor or tumor-cell by RBC-C3bRR and DTER.Results Comparing the mean value of the number of CR1 on each RBC of high and middle groups with control groups,the mean value of the number of CR1,RBC-C3bRR and DTER of Asparagus officinalis polysaccharide groups are increased significantly.Conclusions Asparagus officinalis polysaccharide can improve the erythrocyte function of S180 mice,which may be one of its most important antitumor mechanisms.
基金supported by the National Key Research and Development Program of China(2019YFA0508502 to XL)National Natural Science Foundation of China(31930039,31821003,91942303 to XL)+1 种基金the General Financial Grant from the China Postdoctoral Science Foundation(2020M670301 to YX)annual funding from Tsinghua University-Peking University Jointed Center for Life Sciences.
文摘Invasive fungal infections are life-threatening,and neutrophils are vital cells of the innate immune system that defend against them.The role of LTA4H-LTB_(4)-BLT1 axis in regulation of neutrophil responses to fungal infection remains poorly understood.Here,we demonstrated that the LTA4H-LTB_(4)-BLT1 axis protects the host against Candida albicans and Aspergillus fumigatus,but not Cryptococcus neoformans infection,by regulating the antifungal activity of neutrophils.Our results show that deleting Lta4h or Blt1 substantially impairs the fungal-specific phagocytic capacity of neutrophils.Moreover,defective activation of the spleen tyrosine kinase(Syk)and extracellular signal-related kinase(ERK1/2)pathways in neutrophils accompanies this impairment.Mechanistically,BLT1 regulates CR3-mediated,β-1,3-glucan-induced neutrophil phagocytosis,while a physical interaction with CR3 with slight influence on its dynamics is observed.Our findings thus demonstrate that the LTA4H-LTB_(4)-BLT1 axis is essential for the phagocytic function of neutrophils in host antifungal immune response against Candida albicans and Aspergillus fumigatus.
文摘Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein Barr virus The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen activated protein kinases (MAPKs, including ERK, JNK, and p38MAPK), and to highlight the molecular mechanism of CD 4 + cell deletion in AIDS Methods HOS cells (HOS CR2) and HOS CD4 cells (HOS CD4CR2) stably expressing CR2 were established and then identified by FACS and Western blotting Activation and blocking tests of MAPKs were assessed by Western blot Cell proliferation was determined using Cell Titer 96 Aqueous One Solution Reagent Results FACS results showed that the positive rates of HOS CR2 and HOS CD4CR2 cells were greater than 96%, and Western blot showed that the CR2 expression levels on HOS CR2 and HOS CD4CR2 cells were high Activation and blocking tests of MAPKs (ERK, JNK, and p38MAPK) were carried out in HOS CR2, HOS CD4, and HOS CD4CR2 cells The activation of MAPKs in HOS CR2 cells stimulated with PMA (100 ng/ml) and NHS (10%) was identical The activation of MAPKs increased at 5 minutes, reached a peak at 10 minutes, and decreased to baseline within 30 minutes, all in a time dependent manner; the activation of MAPKs was blocked by anti CR2 McAb, PD98059 (inhibitor of ERK), and Wortmanin (inhibitor of PI 3K), respectively In HOS CD4 cells, MAPKs were activated by HIV gp160 In HOS CD4CR2 cells, MAPK activation was induced by HIV gp160, 10% NHS, and HIV gp160+10%NHS; phosphorylation of p38MAPK was dramatically induced by HIV gp160+NHS, and lasted for 1 hour The cell proliferation results showed that HIV gp160 inhibited the proliferation of HOS CD4 and HOS CD4CR2 cells ( P <0 01) and that NHS enhanced the effect of HIV gp160 ( P <0 01) Conclusions The activation of MAPKs is independently mediated by CR2 and that anti CR2 McAb, PD98059, and Wortmanin block the activation of MAPKs, respectively The results of the signal transduction and cell proliferation assays of HOS CD4CR2 cells show that CR2 plays a role in the pathogenesis of HIV infection, especially in the inhibition of CD 4 + cell proliferation
基金Supported by National Natural Sciences Foundation of China,No. 81001067the Ministry of Science and Technology International Cooperation Project, No. 2010DFA31870the AstraZeneca Special Research Foundation for Targeted Therapy of the Wu Jieping Medical Foundation, No. 320.6700.09068
文摘AIM: To investigate the molecular mechanisms underlying the reversal effect of emodin on platinum resistance in hepatocellular carcinoma. METHODS: After the addition of 10 μmol/L emodin to HepG2/oxaliplatin (OXA) cells, the inhibition rate (IR), 50% inhibitory concentration (IC 50 ) and reversal index (IC 50 in experimental group/IC 50 in control group) were calculated. For HepG2, HepG2/OXA, HepG2/OXA/T, each cell line was divided into a control group, OXA group, OXA + fibroblast growth factor 7 (FGF7) group and OXA + emodin group, and the final concentrations of FGF7, emodin and OXA in each group were 5 ng/mL, 10 μg/mL and 10 μmol/L, respectively. Single-cell gel electrophoresis was conducted to detect DNA damage, and the fibroblast growth factor receptor 2 (FGFR2), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) and excision repair cross-complementing gene 1 (ERCC1) protein expression levels in each group were examined by Western blotting. RESULTS: Compared with the IC50 of 120.78 μmol/L in HepG2/OXA cells, the IC 50 decreased to 39.65 μmol/L after treatment with 10 μmol/L emodin; thus, the reversal index was 3.05. Compared with the control group, the tail length and Olive tail length in the OXA group, OXA + FGF7 group and OXA + emodin group were significantly increased, and the differences were statistically significant (P < 0.01). The tail length and Olive tail length were lower in the OXA + FGF7 group than in the OXA group, and this difference was also statistically significant. Compared with the OXA + FGF7 group, the tail extent, the Olive tail moment and the percentage of tail DNA were significantly increased in the OXA + emodin group, and these differences were statistically significant (P < 0.01). In comparison with its parental cell line HepG2, the HepG2/OXA cells demonstrated significantly increased FGFR2, p-ERK1/2 and ERCC1 expression levels, whereas the expression of all three molecules was significantly inhibited in HepG2/ OXA/T cells, in which FGFR2 was silenced by FGFR2 shRNA. In the examined HepG2 cells, the FGFR2, p-ERK1/2 and ERCC1 expression levels demonstrated increasing trends in the OXA group and OXA + FGF7 group. Compared with the OXA group and OXA + FGF7 group, the FGFR2, p-ERK1/2, and ERCC1 expression levels were significantly lower in the OXA + emodin group, and these differences were statistically significant. In the HepG2/OXA/T cell line that was transfected with FGFR2 shRNA, the FGFR2, p-ERK1/2 and ERCC1 expression levels were significantly inhibited, but there were no significant differences in these expression levels among the OXA, OXA + FGF7 and OXA + emodin groups. CONCLUSION: Emodin markedly reversed OXA resistance by enhancing OXA DNA damage in HepG2/OXA cells, and the molecular mechanism was related to the inhibitory effect on ERCC1 expression being mediated by the FGFR2/ERK1/2 signaling pathway.
