Effect of Xiaoyan Decoction on bone marrow suppression of mice with Lewis lung cancer / 中草药

Objective: To study the effects and mechanism of Xiaoyan Decoction on bone marrow suppression in mice from aspects of peripheral blood cell ratio, immune function, and cytokines. Methods: A mouse model of bone marrow suppression was established. Fifty mice were randomly divided into five groups (n = 10): control group, model group, Xiaoyan Decoction group, rhG-CSF group, and Xiaoyan Decoction combined with rhG-CS group. Lewis-tumor mice in Xiaoyan Decoction group were ig given Xiaoyan Decoction 18.2 g/(kg·d) twice daily, for 7 d. Lewis-tumor mice in rhG-CS group were treated by sc injection of rhG-CSF. The routine blood test, immunological functions of peripheral blood and cytokines (IL-3, IL-6, EPO, and GM-CSF) in serum were detected. Results: The bone marrow suppression model was successfully established. Compared with control group, the levels of WBC, RBC, Hb, and PLT in peripheral blood of model group were decreased, especially in WBC (P 0.05). The concentration of IL-3 and IL-6 was dramatically reduced in model group compared with control group (P < 0.05). And it showed an obviously increasing trend in IL-3, IL-6, EPO, and GM-CSF levels in Xiaoyan Decoction group and combined group, as well as an increase in IL-3, IL-6 levels in rhG-CS group (P < 0.05). Conclusion: It shows that Xiaoyan Decoction combined with rhG-CS can improve the level of WBC and immunological functions of peripheral blood in bone marrow suppression model mice. And the effect of combination of both drugs is more significant. This might be related to the effect of promoting the production of serum hematopoietic regulator such as IL-3, IL-6, EPO, and GM-CSF, and relieving chemotherapy-induced bone marrow suppression.

Regulatory effects of CD44 antibody-A3 D8 on IL-3 Rαand downstream PI3 K/Akt pathway in NB4 cells / 中国药理学通报

Aim To investigate the effect of CD44 anti-body-A3 D8 on the expression of IL-3 Rα and down-stream PI3K/Akt in NB4 cells. Methods The ex-pression of IL-3 Rα mRNA was detected by real-time quantitative RT-PCR, the IL-3Rα protein expression and changes of PI3 K/Akt signal pathway in NB4 cells treated with A3D8 were analyzed by Western blot. An-nexin-V-FITC/PI double staining flow cytometry was u-tilized to detect the apoptotic cells. The inhibitor of PI3 K/Akt signaling LY294002 combined with A3 D8 was used to inhibit the PI3K/Akt in NB4 cells. Re-sults After treated with A3 D8 , both the transcription-al level and translational level of IL-3 Rα were remark-ably reduced, and the PI3K/Akt pathway was inhibi-ted. LY294002 improved the inhibitory and apoptotic effects of A3D8 on NB4 cells. Conclusion CD44 antibody A3 D8 can downregulate the expression of IL-3Rα and inhibit the downstream PI3K/Akt pathway.

Differential Expressions of IL-3 and FcepsilonRI in the Peripheral Blood Mononuclear Cells and Lesional Skin of Patients with Extrinsic Atopic Dermatitis and Intrinsic Atopic Dermatitis / 대한피부과학회지

BACKGROUND: A small subgroup of atopic dermatitis (AD) patients show low total and allergen-specific immunoglobulin (IgE) levels. This subgroup has been termed 'intrinsic' AD (IAD) as compared to its counterpart 'extrinsic' AD (EAD). However, the difference of cytokine expression between IAD and EAD has not been fully understood. OBJECTIVE: To compare the expression of various inflammatory cytokines in the peripheral blood mononuclear cells (PBMCs) and lesional skin of patients with IAD and EAD, which are known to be associated with AD pathophysiology. METHODS: We assessed the protein levels of cytokines in the PBMCs and lesional skin. We evaluated the levels of IL-3, IL-4, IL-5, IL-6, IL-10, IL-13, FcepsilonRI and FcepsilonRII from the PBMCs and lesional skin of patients with IAD and EAD. RESULTS: The patients with EAD had elevated levels of the IL-3 expression in their PBMCs and elevated levels of FcepsilonRI in their lesional skin compared to that of the patients with IAD. The expression of other cytokines did not differ in the PBMCs and lesional skin from the two subgroups. CONCLUSION: This study suggests that IL-3 could be associated with the pathophysiology of EAD as compared to that of IAD, along with FcepsilonRI which was previously shown to be highly expressed in EAD patients.

Effects of IL-3 and SCF on Histamine Production Kinetics and Cell Phenotype in Rat Bone Marrow-derived Mast Cells

BACKGROUND: Rat mast cells were regarded as a good model for mast cell function in immune response. METHODS: Rat bone marrow mast cells (BMMC) were prepared both by recombinant rat IL-3 (rrIL-3) and by recombinant mouse stem cell factor (rmSCF), and investigated for both proliferation and differentiation in time course. Rat BMMC was induced by culture of rat bone marrow cells (BMCs) in the presence of both rrIL-3 (5 ng/ml) and rmSCF (5 ng/ml). Culture media were changed 2 times per week with the cell number condition of 5x10(4)/ml in 6 well plate. Proliferation was analyzed by cell number and cell counting kit-8 (CCK-8) and differentiation was by rat mast cell protease (RMCP) II and histamine. RESULTS: Cell proliferation rates reached a maximum at 8 or 11 days of culture and decreased thereafter. However, both RMCP II production and histamine synthesis peaked after 11 days of culture. By real time RT-PCR, the level of histidine decarboxylase mRNA was more than 500 times higher on culture day 11 than on culture day 5. By transmission electron microscopy, the cells were heterogeneous in size and contained cytoplasmic granules. Using gated flow cytometry, we showed that cultured BMCs expressed high levels of FcepsilonRI and the mast cell antigen, ganglioside, on culture day 11. CONCLUSION: These results indicate that rat BMMCs were generated by culturing BMCs in the presence of rrIL-3 and rmSCF and that the BMMCs have the characteristics of mucosal mast cells.

Construction and prokaryotic expression of fusion gene IL3-PE38KDEL / 重庆医科大学学报

Objective:To construct a prokaryotic expression plasmid of PQE30-IL3-Linker-PE38KDEL and identify its recombinant protein expression.Methods:The IL3 and PE38KDEL gene were amplified by polymerase chain reaction(PCR) and cloned into the prokaryotic expression plasmid PQE30-Linker constructed after being sequenced.The recombinant vector confirmed by restriction endonucleases digestion,coenobium PCR,and DNA sequence analysis was transformed into E.coli SG13009.The expression of the protein was induced by IPTG.Relative molecular weight of the expression product was detected by SDS-PAGE.Finally,the fusion protein was examined by Western blot.Results:The results of restriction endonuclease digestion,coenobium PCR and DNA sequence analysis showed that the prokaryotic expression vector PQE30-IL3-Linker-PE38KDEL was constructed successfully.With induction of IPTG,the relative molecular weight of the expression product was identical to the expected value.The expressed 6?His-IL3-PE38KDEL fusion protein were identified at relative molecular mass of 57KD by Western blot with anti-His monoclonal antibody,showing the fusion protein expressed correctly.Conclusion:The fusion protein IL3-PE38KDEL is successfully constructed,which lays a solid foundation for the further research of protein purification and function.

Prevention of IL-3 Deprivation Induced Apoptosis in Mast Cells by LPS / 대한해부학회지

Mast cells play a protective role in host defense against bacteria, and are found in high number at the host-environment interface. Stimulation of mast cells by bacterial cell wall components is essential for their protective effects against entero-bacterial infection. Mast cells have an extraordinarily long longevity in peripheral tissues compared to other types of blood cells. We undertook this study to reveal the mechanism underlying the prevention on IL-3 deprivation-induced apoptosis in mast cells by LPS. Bone marrow derived mast cells (BMMCs) were obtained from femurs of BALB/c mice and cultured under IL-3 stimulation for 4-6 weeks. At the same point when IL-3 was depleted, BMMCs were treated with LPS. To examine the effect of LPS, DNA electrophoresis, Western blotting, immunoflourescense staining, confocal microscopy, RT-PCR and immunoprecipitation were conducted. IL-3 deprivation reduced cellular viability and induced nuclear condensation and translocation of apoptosisassociated factors. However, LPS treatment prevented these IL-3 deprivation-induced apoptotic events. IL-3 deprivation downregulated representative antiapoptotic factors such as XIAP, 14-3-3, Bcl-2 and Bcl-xL. Expression level of these factors was maintained in BMMCs treated by LPS, which was similar to the level in the control cells. The Bim to Bcl-2 interaction was increased in IL-3 depleted cells, which was prevented by LPS treatment. It was also demonstrated that LPS induced the new expression of a cytokine-induced antiapoptotic factor anamorsin gene in BMMCs under IL-3 deprived condition. Taken together, LPS prevents IL-3 deprivation induced apoptosis in BMMCs via several antiapoptotic factors. This preventive mechanism of LPS on BMMCs apoptosis may contribute the long longevity of mast cells in the peripheral tissue.

Effects of Intermittent Hypoxic Exposure on the Production of Erythrocyte and Some Haematopoietic Factor in Rats with Exercise-Induced Anemia / 中国运动医学杂志

0.05).Conclusion The hypoxic exposure can accelerate the production of erythrocyte, and treat the exercise-induced anemia effectively. Improvement of some haematopoietic factors and enhancement of hematopoiesis in marrow is thought to be the possible mechanism.

Studies of cocktail therapy with multiple cytokines for neoplasia or infectious disease of the dog I. cDNA cloning of canine IL-3 and IL-6

This paper describes the cloning and sequence analysis of the cDNAs encoding the canine homologues of interleukin-3 (IL-3) and interleukin-6 (IL-6). The coding sequences for canine IL-3 and IL-6 were obtained by using the reverse transcription polymerase chain reaction (RT-PCR) with RNA harvested from canine peripheral blood mononuclear cells (PBMCs). Canine IL-3 cDNA includes a single open reading frame of 432 nucleotides, which encodes a 143 amino acid polypeptide and has 44.7, 42.4, 37 and 23.7% homology with the cow, sheep, human and rat IL-3 sequences, respectively. Canine IL-6 cDNA (GenBank accession number; AF275796) encodes a putative 20-amino acid signal peptide followed by a 187-amino acid mature protein. The predicted amino acid sequence of canine IL-6 shares 60.4, 77.2, 71.0, 55.8 and 42.0% sequence identity with those of human, feline, porcine, sheep and rat IL-6, respectively.

The role of cytokines in rhinosinusitis

Since the last decade, new insights into inflammatory processes have become possible by investigating the pattern of cytokines in acute and chronic sinus diseases. This review aims to update and discuss the findings of in vitro and in vivo studies concerning the role of cytokines in sinusitis and nasal polyposis. The proinflammatory cytokines interleukin-1beta, interleukin-6 and the neutrophil-chemoattractant interleukin-8 may play a major role in acute sinusitis, as shown in viral and allergic rhinitis. In chronic sinusitis interleukin-3 dominates the cytokine profiles, giving support to a variety of inflammatory cells. Interleukin-5 is a key protein in the pathogenesis of nasal polyposis. Activation and survival of eosinophils in nasal polyps are thought to be regulated by interleukin-5. Further investigation of cytokine expression patterns in inflammatory sinus diseases will lead to a better understanding of their pathogenesis and to a development of new therapeutic modality.

The role of cytokines in rhinosinusitis

Since the last decade, new insights into inflammatory processes have become possible by investigating the pattern of cytokines in acute and chronic sinus diseases. This review aims to update and discuss the findings of in vitro and in vivo studies concerning the role of cytokines in sinusitis and nasal polyposis. The proinflammatory cytokines interleukin-1beta, interleukin-6 and the neutrophil-chemoattractant interleukin-8 may play a major role in acute sinusitis, as shown in viral and allergic rhinitis. In chronic sinusitis interleukin-3 dominates the cytokine profiles, giving support to a variety of inflammatory cells. Interleukin-5 is a key protein in the pathogenesis of nasal polyposis. Activation and survival of eosinophils in nasal polyps are thought to be regulated by interleukin-5. Further investigation of cytokine expression patterns in inflammatory sinus diseases will lead to a better understanding of their pathogenesis and to a development of new therapeutic modality.

Antagonism of new Flavonoids from Sophorica japonica on IL-3-induced eosinophil activation / 천식및알레르기

BACKGROUND: Even though allergic disease is an inflammatory disease, main inflammatory cells and cytokines involved in this process are different from those in other inflammatory diseases. New therapeutic targets on allergic inflammation include eosinophils, mast cells, T lymphocytes and their cytokines activating those cells, so new antagonists acting on them are under many investigations. We extracted four iso-flavonoids from Sophorica japonica such as Sophi, Orbol, Luten, and Genistein which has been known PTK antagonist. We documented the three iso-flavonoids, except Genistein, had an antagonism on IL-5 using in vitro IL -5-induced eosinophil activation model and also in allergic mouse model sensitized by OA (ovualbumin). One of our data from mouse model, which was those three compounds blocked early allergic response near completely, suggested they might have an antagonism on IL-3, the major cytokine activating mast cell and basophils which control early allergic response mainly. OBJECTIVE: From the above results, we tried to find antagonistic effects of the compounds on IL-3 using IL-3 - induced eosinophil activation model in vitro. METHODS: LTC4 by RIA and degree of degranulation by light and electron microscopic examination were used as the activation markers of eosinophils. RESULTS: Among the compounds, Sophi was the most potent antagonist on IL-3 which induced LTC4 release and even on degranulation, and Orbol and Luten also had antagonism on them, but Genistein, an antagonist of PTK didn't show any antagonistic effects. CONCLUSION: We conclude that those three iso-flavonoids were IL-3 antagonists, and its mechanism might not be through PTK signaling.

The Effect of rhGM-CSF/IL-3 Fusion Protein on Apoptosis of HL-60 Cells Mediated by Ara-C / 中国肿瘤生物治疗杂志

The effect of rhGM-CSF/IL-3 on apoptosis of Ara-C-induced myeloid leukemic cell line HL- 60 was investigated. The results indicated that treatment with rhGM-CSF/IL-3 in combination with Ara-C significantly inhibited the colony growth of HL-60 and enhanced the oligonucleosomal DNA fragmentation as compared with Ara-C alone. The Ara-C mediated apoptosis rates of HL-60 cells treated with rhGM-CSF/IL-3 fusion protein alone were markablely improved compared with treatment with rhIL-3 plus rhGM-CSF, it was noted that the Ara-C mediated of apoptosis normal peripheral white blood cells was less affected by rhGM-CSF/IL-3. It suggested that rhGM-CSF/IL-3 could be used as a possible curing drug during the phase of induced remission of leukemia.

The Effect of Several Cykokines to Growth of Mast Cells / 中国免疫学杂志

Using in vitro cultured mouse bone marrow mast cells as experimental model,effects of sev-eral cytokines to the growth of mast cells were observed.The results show that mSCF can sup-port the growth of long term cultured homogenous mast cells,but cannot stimulate the initiativegrowth of primary cultured bone marrow cells IL—3 and Epo can also support growth of mastcells.