ABSTRACT
Objective To study the effect of Wulong granules on the expression of IL-2 mRNA,IL-4 mRNA and IL-6 mRNA in allergic rhinitis(AR) rats in Spleen Qi deficiency type. To establish the clinic treatment principle-tonify Spleen and reinforce Qi of AR through the lab study. Methods Established AR model with combination of disease and Zheng,checked the expression of IL-2 mRNA,IL-4 mRNA and IL-6 mRNA of nasal mucosa were detected with RT-PCR. Results Wulong granules can adjust the expression of mRNA of AR rats' nasal mucosa. It can descend the ascensus of IL-4 mRNA,IL-6 mRNA and the drop of IL-2 mRNA. Conclusion Wulonggranules can show treatment effect to AR rats through adjusting the expression of cell factors such as IL-2 mRNA,IL-4 mRNA and IL-6 mRNA.
ABSTRACT
PURPOSE: Umbilical cord blood (CB) transplantation is an alternative method instead of allogeneic bone marrow (BM) transplantation. CB transplantation has used grafts with red blood cell (RBC) depletion but previous reports in CB investigated the immune reaction about mononulcear cell separated by density gradient. To study the real immune response in CB transplantation, the experimental group designed the total nuclear cell (TNC) groups by RBC depletion and the mononuclear cell (MNC) groups by Ficoll Hypaque in CB and BM. METHODS: We evaluated the various cytokine gene expression by semiquantitative RT- PCR method after immune stimulation with phytohemagglutinin in cord blood and bone marrow according to cell separation method. RESULTS: 1) All samples of CB and BM expressed IL-2 mRNA. There was no difference in amounts of IL-2 mRNA between CBTNC and CBMNC, between BMTNC and BMMNC. 2) All samples of CB and BM expressed IL-10 mRNA. There was no difference in amounts of IL-10 mRNA between the CBTNC and CBMNC but significantly different between the BMTNC and BMMNC (P<0.05). The amounts of IL-10 mRNA in the BMMNC and CBTNC group showed larger than in the BMTNC group (P<0.05). 3) The expression of IL-4, IFN-gamma was not shown in this study. These results suggest that no difference of IL-2 mRNA between CB and BM may reveal IL-2 as major cytokine gene in CB after immune stimulation. The expression of IL-10 mRNA in CBTNC showed more than in BMTNC group. Conclusion: Our results suggest that CBTNC may contain a lot of cells producing IL-10, as a cytokine of immunoregulatory function, and therefore the immune reaction in CB transplantation may be less apparent than in BM transplantation. The cell component producing IL-10 in CB may be T regulator cell. There were not showed the IFN-gamma and IL-4 mRNA due to short duration of immune stimulation. Therefore, further studies will identify the IL-4 and IFN-gamma mRNA expression after long time stimulation and various cytokine gene expression to certificate the precise immune response after flow cytometry to composed of immune cells in CB and BM.
Subject(s)
Bone Marrow , Cell Separation , Cellular Structures , Diatrizoate , Erythrocytes , Fetal Blood , Ficoll , Flow Cytometry , Gene Expression , Interleukin-10 , Interleukin-2 , Interleukin-4 , Polymerase Chain Reaction , RNA, Messenger , TransplantsABSTRACT
Large scale preparation of human IL-2 mRNA was obtained from human spleen cells stimulated with PHA+TPA at the peak of IL-2 mRNA transcription. Full length of IL-2 mRNA was translated in wheat germ extract to produce bioactive IL-2 (tIL-2). The IL-2 mRNA was about 14 S depending on the 7 M Urea-SDS-PAGE, and recovried from gel and translated in vitro. The elute curve of tIL-2 was the same as that of recombinant IL-2 (rIL-2) and natural IL-2 (nIL-2), the molecular weight of tIL-2 was about 14.000 dalton. The translational process of IL-2 mRNA in vitro was inhibited when RNA was cultured with RNase or translated product was cultured with proteinase, proving a standard in-vitro translation.
ABSTRACT
This study was designed to determine if the decrease in the expression of interleukin 2(IL 2) in spleen lymphocytes after trauma was associated with the changes in the transcription factor NFAT binding activity. Closed impact injury with fracture of both hind limbs in mice was adopted as the trauma model. Animals were sacrificed at the 12h and 1,4,7,10,14 days after injury. Spleen lymphocytes were isolated from traumatized mice and stimulated with concanavalin A, The culture supernatants were harvested and assayed for IL 2 activity. Total RNA was extracted from spleen lymphocytes and assayed for IL 2 mRNA. Nuclear protein was extracted,and DNA binding activity of NFAT was measured using an electrophoretic mobility shift assay(EMSA). The results showed that the DNA binding activity of NFAT gradually decreased , with trough value of NFAT binding activity ,accounting for 41% of the control, observed at 4d after injury. It was closely associated with the decline of IL 2 activity and IL 2 mRNA. These data suggest that the decline in the induction of IL 2 expression is,at least, partially due to the impairment in the activation of NFAT in traumatized mice. Taken together, the results are significant to clarify the mechanism of the decline in the induction of IL 2 expression after trauma.