AIM: To examine the factor(s) involved in differentiation of intestinal macrophages (IMACs) using a recently established in vitro model. METHODS: To test whether soluble or membrane bound factors induce IMAC-different...AIM: To examine the factor(s) involved in differentiation of intestinal macrophages (IMACs) using a recently established in vitro model. METHODS: To test whether soluble or membrane bound factors induce IMAC-differentiation, freshly elutriated monocytes (MO) were incubated with conditioned media or cell membranes of intestinal epithelial cells (IEC) or cultured with IEC in transwell systems. To determine the importance of an active migration of MO, three- dimensional aggregates from a 1:1-mixture of MO and IEC were examined by immunohistochemistry and flow cytometry. Apoptosis was examined by caspase-3 Western blots. Extracellular matrix production in differentiation models was compared by immunohistochemistry. RESULTS: IMAC differentiation was observed in a complex three-dimensional co-culture model (multicellular spheroid, MCS) with IEC after migration of MO into the spheroids. By co-culture of MO with conditioned media or membrane preparations of IEC no IMAC differentiation was induced. Co-culture of MO with IEC in transwell- cultures, with the two cell populations separated by a membrane also did not result in intestinal-like differentiation of MO. In contrast to IEC-spheroids with immigrating MO in mixed MCS of IEC and MO only a small subpopulation of MO was able to survive the seven day culture period. CONCLUSION: Intestinal-like differentiation of MO in vitro is only induced in the complex three-dimensional MCS model after immigration of MO indicating a roleof cell-matrix and/or cell-cell interactions during the differentiation of IMACs.展开更多
Objective: To study the expression of MCP-1 in colorectal carcinoma and its relationship to the infiltration of the macrophage and to the biological behaviour of infiltration and metastasis of colorectal carcinoma. Me...Objective: To study the expression of MCP-1 in colorectal carcinoma and its relationship to the infiltration of the macrophage and to the biological behaviour of infiltration and metastasis of colorectal carcinoma. Methods: The expression of the MCP-1 mRNA was assessed in colorectal carcinoma collected freshly from surgical specimen by RT-PCR and the expres- sion of the MCP-1 protein was assessed in colorectal carcinoma collected from surgical specimen by immunohistochemistry. The tumor infiltrating cell and macrophage were also investigated by immunohistochemistry. Results: All the 12 specimens of colorectal carcinoma detected by RT-PCR expressed the MCP-1 mRNA; MCP-1 protein was detected in 90℅ (36/40) cases of the tumor; The expression of the MCP-1 protein in colorectal carcinoma correlated negatively with its state of metastasis and the Dukes’ stage. But a postive correlation was found between the expression of MCP-1 and the infiltrated macrophage. The stron- ger expression of MCP-1, the more number of the infiltrated macrophage. Conclusion: The expression of chemokine MCP-1 in colorectal carcinoma may influence its biological behaviour of infiltration and metastasis, and can attract the immuno-cell to the local of the tumor, such as Macrophage.展开更多
AIM： To investigate the selective cytotoxic effect of constructed hybrid protein on cells expressing granulocyte macrophage colony stimulating factor （GM-CSF） receptor. METHODS： HepG2 （human hepatoma） and LS174T...AIM： To investigate the selective cytotoxic effect of constructed hybrid protein on cells expressing granulocyte macrophage colony stimulating factor （GM-CSF） receptor. METHODS： HepG2 （human hepatoma） and LS174T （colon carcinoma） were used in this study. The fused gene was induced with 0.02% of arabinose for 4 h and the expressed protein was detected by Western blotting. The chimeric protein expressed in E..coli was checked for its cytotoxic activity on these cells and apoptosis was measured by comet assay and nudear staining. RESULTS： The chimeric protein was found to be cytotoxic to the colon cancer cell line expressing GM-CSFRs, but not to HepG2 lacking these receptors. Maximum activity was observed at the concentration of 40 ng/mL after 24 h incubation. The IC50 was 20 ± 3.5 ng/mL. CONCLUSION： Selective cytotoxic effect of the hybrid protein on the colon cancer cell line expressing GMCSF receptors （GM-CSFRs） receptor and apoptosis can be observed in this cell line. The hybrid protein can be considered as a therapeutic agent.展开更多
Objective： To observe the regulatory effect of moxibustion on the expression of CD86 and CD163, which are the important functional phenotypes of macrophage differentiation in lung tissue of ulcerative colitis（UC） r...Objective： To observe the regulatory effect of moxibustion on the expression of CD86 and CD163, which are the important functional phenotypes of macrophage differentiation in lung tissue of ulcerative colitis（UC） rats. The mechanism of the regulatory effect of moxibustion for macrophage differentiation based on the key cytokine interferon-gamma（IFN-γ）, tumor necrosis death factor-alpha（TNF-α）, interleukin- 4（IL-4） and interleukin-13（IL-13） was also explored. Methods： Forty SD rats were randomly divided into a normal group（NG）, a model group（MG）, a normal moxibustion group（NMG） and a smokeless moxibustion group（SMG）. The model of UC was made by antigen immunization combined with enema with topical formalin. The rats in the normal moxibustion group accepted moxibustion at bilateral Tianshu（ST 25）, while the rats in the smokeless moxibustion group with smokeless moxibustion at bilateral Tianshu（ST 25）, each 10 min, once a day for 8 d. After treatments, the hematoxylin-eosin（HE） staining was used to observe the pathological changes of colonic tissue, the Western blotting（WB） was used to observe the expression of CD86 and CD163, two important functional phenotypes of macrophage differentiation in lung tissue of UC rats, while the enzyme-linked immunosorbent assay（ELISA） was used to assess the content of IFN-γ, TNF-α, IL-4 and IL-13, the key cytokines of macrophage differentiation in micro environment of the lung tissue. Results： Compared with the NG, the colons of rats in MG were injured more seriously, and the scores of gross observation and histological examination were significantly higher（P〈0.05）. Compared with the MG, the pathological changes of the two groups of rats with moxibustion treatment were improved, which presented with ulcer repair, inflammation dissipation, and the general score and histological score of the two groups were decreased（P〈0.05）. Compared with the NG, the expression of CD86 in the lung tissue of rats in the MG was increased（P〈0.05）, and CD163 expression was decreased（P〈0.05）. Compared with the MG and the SMG, the expression of CD86 in the lung tissue of the rats in the NMG was significantly lower than those in the MG and SMG, and CD163 was higher（P〈0.05）, while the differences were not statistically significant between the MG and the SMG（P〈0.05）. Compared with the NG, the expression of key cytokines in lung tissue of MG was abnormal, the contents of IFN-γ and TNF-α increased（P〈0.05）, while the IL-4 and IL-13 decreased（P〈0.05）. The IL-4 and IL-13 were significantly increased in lung tissue of rats in the NMG（P〈0.05）, and the IFN-γ and TNF-α were reduced（P〈0.05）, compared with those in the MG and the SMG, while the differences were not statistically significant between the MG and the SMG（P〈0.05）. Conclusion： Moxibustion can increase the expression of CD163, the important functional phenotype of macrophages in lung tissue of UC rats, and the differentiation critical cytokines IL-4 and IL-13. It can also reduce activated phenotype CD86 and its differentiation critical cytokines IFN-γ and TNF-α in the lung tissue of UC rats.展开更多
基金Supported by the Deutsche Forschungsgemeinschaft (SFB585, Ro 1236/3-2)the BMBF Kompetenznetz-CED
文摘AIM: To examine the factor(s) involved in differentiation of intestinal macrophages (IMACs) using a recently established in vitro model. METHODS: To test whether soluble or membrane bound factors induce IMAC-differentiation, freshly elutriated monocytes (MO) were incubated with conditioned media or cell membranes of intestinal epithelial cells (IEC) or cultured with IEC in transwell systems. To determine the importance of an active migration of MO, three- dimensional aggregates from a 1:1-mixture of MO and IEC were examined by immunohistochemistry and flow cytometry. Apoptosis was examined by caspase-3 Western blots. Extracellular matrix production in differentiation models was compared by immunohistochemistry. RESULTS: IMAC differentiation was observed in a complex three-dimensional co-culture model (multicellular spheroid, MCS) with IEC after migration of MO into the spheroids. By co-culture of MO with conditioned media or membrane preparations of IEC no IMAC differentiation was induced. Co-culture of MO with IEC in transwell- cultures, with the two cell populations separated by a membrane also did not result in intestinal-like differentiation of MO. In contrast to IEC-spheroids with immigrating MO in mixed MCS of IEC and MO only a small subpopulation of MO was able to survive the seven day culture period. CONCLUSION: Intestinal-like differentiation of MO in vitro is only induced in the complex three-dimensional MCS model after immigration of MO indicating a roleof cell-matrix and/or cell-cell interactions during the differentiation of IMACs.
基金Supported by the Scientific Research Fundation of the Education Department of Fujian province (No. JA00205).
文摘Objective: To study the expression of MCP-1 in colorectal carcinoma and its relationship to the infiltration of the macrophage and to the biological behaviour of infiltration and metastasis of colorectal carcinoma. Methods: The expression of the MCP-1 mRNA was assessed in colorectal carcinoma collected freshly from surgical specimen by RT-PCR and the expres- sion of the MCP-1 protein was assessed in colorectal carcinoma collected from surgical specimen by immunohistochemistry. The tumor infiltrating cell and macrophage were also investigated by immunohistochemistry. Results: All the 12 specimens of colorectal carcinoma detected by RT-PCR expressed the MCP-1 mRNA; MCP-1 protein was detected in 90℅ (36/40) cases of the tumor; The expression of the MCP-1 protein in colorectal carcinoma correlated negatively with its state of metastasis and the Dukes’ stage. But a postive correlation was found between the expression of MCP-1 and the infiltrated macrophage. The stron- ger expression of MCP-1, the more number of the infiltrated macrophage. Conclusion: The expression of chemokine MCP-1 in colorectal carcinoma may influence its biological behaviour of infiltration and metastasis, and can attract the immuno-cell to the local of the tumor, such as Macrophage.
文摘AIM： To investigate the selective cytotoxic effect of constructed hybrid protein on cells expressing granulocyte macrophage colony stimulating factor （GM-CSF） receptor. METHODS： HepG2 （human hepatoma） and LS174T （colon carcinoma） were used in this study. The fused gene was induced with 0.02% of arabinose for 4 h and the expressed protein was detected by Western blotting. The chimeric protein expressed in E..coli was checked for its cytotoxic activity on these cells and apoptosis was measured by comet assay and nudear staining. RESULTS： The chimeric protein was found to be cytotoxic to the colon cancer cell line expressing GM-CSFRs, but not to HepG2 lacking these receptors. Maximum activity was observed at the concentration of 40 ng/mL after 24 h incubation. The IC50 was 20 ± 3.5 ng/mL. CONCLUSION： Selective cytotoxic effect of the hybrid protein on the colon cancer cell line expressing GMCSF receptors （GM-CSFRs） receptor and apoptosis can be observed in this cell line. The hybrid protein can be considered as a therapeutic agent.
基金supported by National Natural Science Foundation of ChinaNational Basic Research Program of China(973 Program)Special Scientific Research Fund for Selection and Cultivation of Elite in College and University~~
文摘Objective： To observe the regulatory effect of moxibustion on the expression of CD86 and CD163, which are the important functional phenotypes of macrophage differentiation in lung tissue of ulcerative colitis（UC） rats. The mechanism of the regulatory effect of moxibustion for macrophage differentiation based on the key cytokine interferon-gamma（IFN-γ）, tumor necrosis death factor-alpha（TNF-α）, interleukin- 4（IL-4） and interleukin-13（IL-13） was also explored. Methods： Forty SD rats were randomly divided into a normal group（NG）, a model group（MG）, a normal moxibustion group（NMG） and a smokeless moxibustion group（SMG）. The model of UC was made by antigen immunization combined with enema with topical formalin. The rats in the normal moxibustion group accepted moxibustion at bilateral Tianshu（ST 25）, while the rats in the smokeless moxibustion group with smokeless moxibustion at bilateral Tianshu（ST 25）, each 10 min, once a day for 8 d. After treatments, the hematoxylin-eosin（HE） staining was used to observe the pathological changes of colonic tissue, the Western blotting（WB） was used to observe the expression of CD86 and CD163, two important functional phenotypes of macrophage differentiation in lung tissue of UC rats, while the enzyme-linked immunosorbent assay（ELISA） was used to assess the content of IFN-γ, TNF-α, IL-4 and IL-13, the key cytokines of macrophage differentiation in micro environment of the lung tissue. Results： Compared with the NG, the colons of rats in MG were injured more seriously, and the scores of gross observation and histological examination were significantly higher（P〈0.05）. Compared with the MG, the pathological changes of the two groups of rats with moxibustion treatment were improved, which presented with ulcer repair, inflammation dissipation, and the general score and histological score of the two groups were decreased（P〈0.05）. Compared with the NG, the expression of CD86 in the lung tissue of rats in the MG was increased（P〈0.05）, and CD163 expression was decreased（P〈0.05）. Compared with the MG and the SMG, the expression of CD86 in the lung tissue of the rats in the NMG was significantly lower than those in the MG and SMG, and CD163 was higher（P〈0.05）, while the differences were not statistically significant between the MG and the SMG（P〈0.05）. Compared with the NG, the expression of key cytokines in lung tissue of MG was abnormal, the contents of IFN-γ and TNF-α increased（P〈0.05）, while the IL-4 and IL-13 decreased（P〈0.05）. The IL-4 and IL-13 were significantly increased in lung tissue of rats in the NMG（P〈0.05）, and the IFN-γ and TNF-α were reduced（P〈0.05）, compared with those in the MG and the SMG, while the differences were not statistically significant between the MG and the SMG（P〈0.05）. Conclusion： Moxibustion can increase the expression of CD163, the important functional phenotype of macrophages in lung tissue of UC rats, and the differentiation critical cytokines IL-4 and IL-13. It can also reduce activated phenotype CD86 and its differentiation critical cytokines IFN-γ and TNF-α in the lung tissue of UC rats.
