ABSTRACT
BACKGROUND:It may be an important approach to avoiding organ transplant rejection by utilizing immature dendritic cells to induce donor-specific immunologic tolerance. OBJECTIVE:To study the effect of sinomenine on the differentiation and maturation of rat bone marrow-derived dendritic celsin vitro. METHODS:Bone marrow-derived dendritic cells were isolated from the rat femur and tibia, and immature dendritic cells were induced by granulocyte-macrophage colony stimulating factor and interleukin-4. On day 7, lipopolysaccharide was added and the cells were cultured to generate mature dendritic cells. Cells were divided into control group and low-, middle- and high-dose sinomenine treatment groups (SNL, SNM, SNH groups). Forty hours later, dendritic cels were harvested, and cell morphology was observed by inverted phase contrast microscope. The expression of CD80 and RT1B was detected by flow cytometry. ELISA was used to detect the expression of interleukin-12. The mixed lymphocyte reaction was used to detect the ability of dendritic cells to stimulate the activation of allogeneic T lymphocytes. RESULTS AND CONCLUSION: (1) Under the inverted microscope, the morphology of mature dendritic cells was observed in the control group; in the SNL group most dendritic cells were visible; in the SNM group, there were partially suspended cells with poor maturation; and in the SNH group, most of the cells were not mature. (2) The expression of CD80 in the control group was significantly lower than that in the SNL, SNM and SNH groups (P < 0.05), and the expression of RT1B was significantly reduced in the SNM and SNH groups than the control group. (3) Compared with the control group, the level of IL-12p70 in the cell supernatant was significantly decreased in the SNM and SNH groups (P < 0.01). (4) The ability of dendritic cells to stimulate T lymphocyte proliferation in the SNM and SNH groups was significantly decreased compared with the control group (P < 0.05). To conclude, sinomenine can inhibit the maturation of dendritic cells.
ABSTRACT
BACKGROUND:Liver regeneration is the key factor influencing the prognosis of living donor liver transplantation. There has not been the research on special miRNA of liver regeneration after living donor liver transplantation. OBJECTIVE:To analyze the variation of miRNAs expression profile after rat reduced-size liver transplantation at certain time point, select and verify target miRNA which can provide targeting intervention strategies in liver regeneration after rat reduced-size liver transplantation and provide theoretical evidence for liver regeneration after living donor liver transplantation.METHODS:The reduced-size liver transplantation models were established. miRNAs microarray was used to detect miRNA expression. In differentialy expressed microRNAs, real-time quantitative PCR was utilized to detect target miRNAs. The credibility of miRNAs microarray results was verified. RESULTS AND CONCLUSION:Compared with rat liver tissue in the sham operation group, 11 miRNAs up-regulated in reduced-size liver transplantation, including let-7b-5p, let-7c-5p, miR-101a-3p, miR-103-3p, miR-130a-3p, miR-142-5p, miR-186-5p, miR-199a-3p, miR-21-5p, 221-3p and miR-34a-5p. Four miRNAs were down-regulated, including miR-26b-5p, miR-150-5p, miR-19a-3p and rno-miR-146-5p. PCR test further verified that miR-221-3p and miR-199a-3p expression changes approximated the chip results at 24, 48 hours and 1 week, indicating that results of miRNA microarray were believable. These results verified that it exists variation of miRNAs expression profile after rat reduced-size liver transplantation, which picked out and verified the target miRNAs.