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Objective@#To observe the effect of meisoindigo on apoptosis and proliferation of JAK2/V617F heterozygous mutation cell line-SET2 cell line to further explore the role of JAK-STAT pathway in this effect.@*Methods@#Cell apoptosis after treated with different concentration of meisoindigo (0, 5, and 10 μmol/L) was evaluated by flow cytometry at different time points (24, 48, 72 h). Cell proliferation with CCK8 test was evaluated at different time points (24, 48, 72, 96 h) after administered with different concentration of meisoindigo (0, 5, 10, and 20 μmol/L). After treatment with different concentration of meisoindigo (0, 5, 10, and 20 μmol/L), SET2 cells were collected after 12 h, and then cultured in incomplete methylcellulose-based medium for clone formation. JAK-STAT signaling pathway and apoptosis related protein by Western blot test were evaluated 12 h after administered with different concentration of meisoindigo (0, 5, 10, and 20 μmol/L).@*Results@#At different time points after treated with meisoindigo, the apoptosis rate of SET2 cell lines increased (P<0.01) with the inhibited proliferation (P<0.01), and the decreased clone formation rate of SET2 cell lines [0 μmol/L meisoindigo: (4.48±1.19)%, 20 μmol/L meisoindigo: (2.55±0.36)%; Dunnett P=0.020] in the presence of augmented concentrations of meisoindigo. At 12 hours after administration with meisoindigo, the reduced expressions of JAK2, P-JAK2, P-STAT1, P-STAT3, P-STAT3, STAT5, the decreased anti-apoptosis proteins BCL-2, BCL-XL and the increased pro-apoptosis protein BID, BIM were observed in the presence of increased concentrations of meisoindigo.@*Conclusion@#Meisoindigo played an important role during the apoptosis and the inhibition of proliferation in ph-negative myeloproliferative neoplasm cell-SET2 cell lines, which might be related to the inhibition of JAK-STAT signaling pathway with up-regulation of pro-apoptosis protein and down-regulation of anti-apoptosis protein.
ABSTRACT
Objective: To observe the effect of meisoindigo on apoptosis and proliferation of JAK2/V617F heterozygous mutation cell line-SET2 cell line to further explore the role of JAK-STAT pathway in this effect. Methods: Cell apoptosis after treated with different concentration of meisoindigo (0, 5, and 10 μmol/L) was evaluated by flow cytometry at different time points (24, 48, 72 h). Cell proliferation with CCK8 test was evaluated at different time points (24, 48, 72, 96 h) after administered with different concentration of meisoindigo (0, 5, 10, and 20 μmol/L). After treatment with different concentration of meisoindigo (0, 5, 10, and 20 μmol/L), SET2 cells were collected after 12 h, and then cultured in incomplete methylcellulose-based medium for clone formation. JAK-STAT signaling pathway and apoptosis related protein by Western blot test were evaluated 12 h after administered with different concentration of meisoindigo (0, 5, 10, and 20 μmol/L). Results: At different time points after treated with meisoindigo, the apoptosis rate of SET2 cell lines increased (P<0.01) with the inhibited proliferation (P<0.01), and the decreased clone formation rate of SET2 cell lines [0 μmol/L meisoindigo: (4.48±1.19)%, 20 μmol/L meisoindigo: (2.55±0.36)%; Dunnett P=0.020] in the presence of augmented concentrations of meisoindigo. At 12 hours after administration with meisoindigo, the reduced expressions of JAK2, P-JAK2, P-STAT1, P-STAT3, P-STAT3, STAT5, the decreased anti-apoptosis proteins BCL-2, BCL-XL and the increased pro-apoptosis protein BID, BIM were observed in the presence of increased concentrations of meisoindigo. Conclusion: Meisoindigo played an important role during the apoptosis and the inhibition of proliferation in ph-negative myeloproliferative neoplasm cell-SET2 cell lines, which might be related to the inhibition of JAK-STAT signaling pathway with up-regulation of pro-apoptosis protein and down-regulation of anti-apoptosis protein.
Subject(s)
Apoptosis , Cell Line , Cell Proliferation , Indoles , STAT3 Transcription FactorABSTRACT
Aim To observe the influence of meisoindi-go on the alteration of Wnt/β-catenin signaling in type 1 diabetic rats ' myocardium and clarify its role in the development of diabetic cardiomyopathy .Methods The type 1 diabetes rat model was established by injec-tion of streptozocin after one-week adaptive feeding . The successful modeling rats were randomly divided in-to DM model group of 4 weeks and 8 weeks, meisoindi-go group of 4 weeks and 8 weeks.Fasting blood glu-cose(FBG) levels were tested.HE staining was used to observe the pathological changes of myocardial struc-tures.The alteration of GSK-3β, p-GSK-3β, Wnt2,β-catenin, NF-κB-p65, p-NF-κB-p65 in myocardium was determined by Western blot and immunohistochem-istry.Results Compared with control group , FBG levels of type 1 diabetic rats significantly increased ( P<0.01 ) , while body weight levels significantly de-creased ( P<0.01 ); compared with DM group , FBG levels of 8 weeks meisoindigo group significantly de-creased ( P <0.01 ) .Myocardial histological analysis revealed that DM induced myocardial focal myocyte hypertrophy , solubility , necrosis , fiber tissue hyperplasi-a; compared with DM group , these symptoms were eased in meisoindigo group of 4 weeks and 8 weeks. Compared with control group , the expression of p-GSK-3β, Wnt2, β-catenin, p-NF-κB-p65 level increased, especially with DM group of 8 weeks(P<0.01).The expression of p-GSK-3β, Wnt2,β-catenin, p-NF-κB-p65 level in meisoindigo group of 4 weeks and 8 weeks decreased significantly(P<0.01).Conclusions The repair effect of meisoindigo on myocardial damage in type 1 diabetic rats may be caused by lowering the ex-pression of proteins in Wnt/β-catenin signaling and in-hibiting the activation of Wnt/β-catenin signaling path-way, participating in the repair of myocardial damage and inflammatory in diabetic rats .Further researches on its mechanism in repairing diabetic myocardial dam-age may find new therapeutic targets for diabetic car-diomyopathy .
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Objective To investigate the effect of meisoindigo inducing primary K562 cell apoptosis and its influence on the expression of conduction signal STAT5.Methods K562 cell was treated with meisoindigo for 12 h、24 h、48 h respectively.Flow cytometry was used to detect cell apoptosis and cell cycle,and Western blot was employed to examine the alteration of the expression of STAT5.Results ①The rate of K562 cell apoptosis treated with meisoindigo for 12 h、24 h、and 48 h was 12.5 ± 0.69,19.4± 1.07 and 42.5 ± 3.22,showing statistical significance compared with the control group (P＜0.05).②Meisoindigo could block K562 cell at the S stage,the ratio of S stage cell was increased to 70.48±6.34 with the decrease of cells at G0/G1 and G2/M stage.③Gradually reduced expression of STAT5 showed after K562 cells treated with meisoindigo from 12h to 48h.Conclusion Meisoindigo accelerrates the apoptosis of K562 cell and the mechanism may relevant to the decreased expression of STAT5.
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Objective To observe the effect of meisoindigo on co-culture leukemic cells and bone marrow stromal cells.Methods Bone marrow stromal cells were cultured using bone marrow mononuclear cells from patients with leukemia,and built the co-culture leukemic cells and bone marrow stromal cells.Trypan-blue-exclusion test detected the proliferation of leukemic cells.The expression levels of membrane CXCR4 on leukemic cells were assessed by flow cytometric analysis.Results Bone marrow stromal cell of leukemia could inhibit leukemic cells proliferation,leukemic cells could be promoted their growth when exposed to low concentration (5 μmol/L) of meisoindigo.Abnormal high expression of CXCR4 in leukemic cells,and meisoindigo could markedly inhibit the expression of CXCR4 in HL-60 and del leukemic cells.Expression of CXCR4 in leukemic cells can up-regulated by bone marrow stromal cells from patients with leukemia.Conclusion Meisoindigo can down regulate expression of CXCR4 in leukeimic cells to antileukemic cells.