摘要
Objective: To analysis the number of spleen B-cells and the expression level of inflammatory factors(TNF-α and IL-1β) and Sirt1 in spleen B-cells of CD38-/- mice, to explore the effects of CD38 gene knockout on inflammatory factors of Bcells and its' potential mechanisms. Methods Sorting B-cells from spleens of wide type(WT) C57BL/6 and CD38-/- mice individually by negative sorting method with magnetic beads, counting the cell number, and identifing the purity of sorting B-cells by flow cytometry. cDNA was amplified by RT-PCR, and protein was extracted by RIPA. The mRNA expression levels of inflammatory factors and CD38 gene were detected by Real-time PCR, the protein expression levels of CD38 gene, Neo gene and Sirt1 gene were detected by Western-blot. Results We isolated B-cells from both WT and CD38-/- mice succesfully(putity>95%), idetified CD38-/- mice, the development of spleen was obstacled, and both the number of spleen cells and B-cells in spleen were significantly decreased in CD38-/- mice(P<0.01), accompanied with the decreasing expression level of TNF-α(P<0.01) and IL-1β(P<0.01), expression level of Sirt1 was significantly increased. Conclusion CD38 gene knockout may inhibits the expression of inflammatory factors(TNF-α and IL-1β) in spleen B-cells by activates Sirt1 pathway, but it's effects in antoimmune diseases need further study.
Objective: To analysis the number of spleen B-cells and the expression level of inflammatory factors(TNF-α and IL-1β) and Sirt1 in spleen B-cells of CD38~(~(-/-)) mice, to explore the effects of CD38 gene knockout on inflammatory factors of Bcells and its' potential mechanisms. Methods Sorting B-cells from spleens of wide type(WT) C57BL/6 and CD38~(-/-) mice individually by negative sorting method with magnetic beads, counting the cell number, and identifing the purity of sorting B-cells by flow cytometry. c DNA was amplified by RT-PCR, and protein was extracted by RIPA. The mRNA expression levels of inflammatory factors and CD38 gene were detected by Real-time PCR, the protein expression levels of CD38 gene, Neo gene and Sirt1 gene were detected by Western-blot. Results We isolated B-cells from both WT and CD38~(-/-) mice succesfully(putity>95%), idetified CD38~(-/-) mice, the development of spleen was obstacled, and both the number of spleen cells and B-cells in spleen were significantly decreased in CD38~(-/-) mice(P<0.01), accompanied with the decreasing expression level of TNF-α(P<0.01) and IL-1β(P<0.01), expression level of Sirt1 was significantly increased. Conclusion CD38 gene knockout may inhibits the expression of inflammatory factors(TNF-α and IL-1β) in spleen B-cells by activates Sirt1 pathway, but it's effects in antoimmune diseases need further study.
引文