Angelica Polysaccharide and TPO Have a Protective Effect On Hcmv-Induced Apoptosis In Megakaryocytes

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3553-3553
Author(s):  
Mo Yang ◽  
Jian Liang Chen ◽  
Jie yu Ye ◽  
Su yi Li ◽  
En yu Liang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Protective Effect ◽  
Caspase 3 ◽  
Hcmv Infection ◽  
Annexin V ◽  
Angelica Sinensis ◽  
Control Group ◽  
Mock Control ◽  
Apoptotic Effect ◽  
Induced Apoptosis

Abstract Human cytomegalovirus (hCMV) infection is often associated with thrombocytopenia. Megakaryocytes may be one of the major sites of hCMV infection, then inducing this cell apoptosis. Angelica Sinensis (Danggui) is an important ingredient of many commonly used herbal Medicine for promoting blood production. Our previous study has showed that the hematopoietic effect of Angelica Sinensis is related to its constituent, angelica polysaccharide (APS) (Yang M et al, J Ethnopharma, 2009). This present study investigated the anti-apoptotic effect of APS and TPO on hCMV-induced apoptosis in megakaryocytes. Human bone marrow mononuclear cells (MNC) or megakaryocytic cell line CHRF-288-11 and hCMV AD169 strain were co-cultured in this study. hCMV significantly inhibited the formation of CFU-MK as shown in three different concentrations of viral infection groups (103, 104 and 105 pfu/ml), compared with blank control and mock control (n=10, P<0.05). hCMV also significantly inhibited the growth of CHRF cells in these three different concentrations after incubation for 3 days, which compared with control group (n=10, P<0.01). hCMV DNA and mRNA were also positively detected in CHRF cells and the cells of CFU-MK with IS-PCR and RT-PCR respectively, while it was negative in blank and mock control groups. We further studied the effect of APS and TPO on CFU-MK formation. Results showed that APS (50 ug/ml) like TPO (50 ng/ml) enhanced hCMV-reduced CFU-MK (P=0.05, n=6). CHRF cells were also analyzed by Annexin V/PI with flow cytometry at day 3 after infection with hCMV AD169. The percentage of apoptotic cells in group of 103 pfu/ml was 19.0 ± 2.0%; The group of 104 pfu/ml was 23.0 ± 1.5%; The group of 105 pfu/ml was 28.0 ± 3.0%. The control group was 2.0 ± 0.5%. The apoptotic cells were confirmed by morphologic observation. In addition, apoptotic signals from megakaryocytic surface, cytoplasma and mitochondria were detected in hCMV infected cells by flow cytometry with Caspase-3 and JC-1 assay. Compared to mock infection control at day 5, Annexin-V positive cells population increased by 58%; active caspase-3 signal increased by 120% in viable cell population; and cell population with damaged mitochondial membrane showed a 5-times increase. Moreover, the anti-apoptotic effect of APS and TPO on CHRF cells was also demonstrated by using Annexin-V assay. Our studies showed that hCMV induces the apoptosis in megakaryocytes via mitochondrial and caspase-3 signaling, and angelica polysaccharide (APS) like TPO has a protective effect on hCMV-induced apoptosis in these cells. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Protective Effect of Thrombopoietin on CoCl2-Induced Apoptosis of HUVEC Cells through PI3-k/Akt Pathways

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4969-4969 ◽  
Author(s):  
Jieyu Ye ◽  
Jun-yan Wang ◽  
En-yu Liang ◽  
Mo Yang
Keyword(s):  
Protective Effect ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Annexin V ◽  
Akt Pathway ◽  
Human Endothelial Cells ◽  
Apoptotic Effect ◽  
Huvec Cells ◽  
Induced Apoptosis ◽  
Active Caspase

Abstract Endothelial damage is a serious syndrome following by stem cell transplantation. However, a useful treatment for attenuating the periods of this disorder was still under explored. Our previous studies have demonstrated that in patients with acute inflammation states, serum thrombopoietin (TPO) levels were significant higher compared with healthy subjects (181.11 ± 35.38 vs 96.13 ± 9.7 pg/ml, p<0.001), which may act as an acute response protein to protect the body. TPO is now recognized as a potent proliferative factor for outer hematopoietic system, such as cardiomyocytes and neurocytes. This study is aim to investigate the protective effect of TPO on human endothelial cells, and to explore its potential mechanism. Four experimental groups were set up using human umbilical vein endothelial (HUVEC) cells, which are: Normal control, CoCl2 alone (800 μmol/L), TPO(100 μg/L) + CoCl2 and TPO alone groups. CoCl2 alone decreased the viability of HUVEC cells in a dose dependent manner (r=-0.997), while addition of TPO significantly rescued this effect (n=5, p<0.01). We further tested the anti-apoptotic effect of TPO on HUVEC cells. CoCl2 alone was able to induced apoptosis as demonstrated by Annexin V (n=4, p<0.001), active-caspase-3 (n=4, p<0.01) and Mitochondrial Membrane potential (JC-1) assays (n=4, p<0.01). Consistently, TPO plus CoCl2 showed a significant anti-apoptotic effect compared with CoCl2 alone group in Annexin V (n=4, p<0.01), active-caspase-3 (n=4, p=0.039) and Mitochondrial Membrane potential (JC-1) assays (n=4, p=0.038). In order to find out the underlying mechanism, we further tested the expression of phospho-Akt (p-Akt). Our results demonstrated that CoCl2 alone suppressed the PI-3k/Akt pathway. TPO plus CoCl2 was shown to activate the expression of p-Akt by western blot. In summary, our findings showed that TPO had a protective effect on human endothelial cells. This effect was likely mediated by activation of PI-3k/Akt pathway, which leads to anti-apoptosis on these cells. This may provide us a new clue to identify novel strategy in treating endothelial syndromes. Disclosures No relevant conflicts of interest to declare.

Iron-Overload Induces Apoptosis in Cardiomyocytes and Hepatocytes Via Mitochondrial/Caspase-3 Pathways.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1872-1872
Author(s):  
Mo Yang ◽  
Shing Chan ◽  
Yiu Fai Cheung ◽  
Shau Yin Ha ◽  
Godfrey ChiFung Chan
Keyword(s):  
Iron Overload ◽  
Protective Effect ◽  
Cell Viability ◽  
Caspase 3 ◽  
Annexin V ◽  
Apoptotic Cells ◽  
H9c2 Cells ◽  
Plot Analysis ◽  
Iron Treatment ◽  
Induced Apoptosis

Abstract Cardiomyopathy and liver damage due to iron-overload are the major complications in patients with beta-thalassaemia major. Iron-overload may induce apoptosis in cardiomyocytes and hepatic cells, and that TPO may exert protective effect on apoptosis of cardiomyocytes (Circulation, 2006). In this study, we demonstrated firstly that iron induced apoptosis in cardiomyocytes. Using H9C2 cells, we have shown that iron reduced cell viability in a dose-dependent manner (0.003–3 mM) (n=6). By annexin V and PI staining, apoptotic cells were found to be significantly increased after iron treatment (0.3 mM, 72 hrs) (n=6). The expression of active caspase-3 was significantly increased in iron-treated cells. Furthermore, iron treatment increased the proportion of cells containing JC-1 monomers, indicating a trend in the drop of mitochondrial membrane potential (n=6). Secondly, we found that TPO exerted cardio-protective effect on iron-induced apoptosis. H9C2 cells were cultured in the presence of iron (0.3 mM) with or without TPO (5, 10, 20, 50, 100 ng/mL, 72 hrs). The cell viability was significantly increased with the treatment of TPO at 50 ng/mL and 100 ng/mL (n=4). Dot-plot analysis of annexin V/PI staining demonstrated that TPO (50 ng/mL) significantly reduced the population of apoptotic cells (n=6). Incubation with TPO also decreased the iron-induced caspase-3 expression (n=6). Flow cytometric dot-plot analysis of H9C2 cells also showed trends of amelioration of the increase in JC-1 monomers in the iron plus TPO group (n=6). The population of phospho-Akt and Erk1/2 were also significantly increased after treatment by TPO (P&lt;0.05, n=4). Human liver cell line MIHA was also used as a cell model. We showed that iron-overload reduced cell viability in a dose-dependent manner (0.0375–0.6 mM) (n=7). By annexin V and PI staining, apoptotic cells were found to be significantly increased after iron treatment (0.15–0.6 mM) for 72 hrs (n=7). The expression of active caspase-3 was also significantly increased in iron-treated cells (n=5). We also found that TPO exerted proliferation effect on MIHA cell by activation of phospho-Akt. However, MIHA cells were cultured in the presence of iron (0.3 mM) with TPO (50 ng/mL, 72 hrs). The cell viability was not significantly increased with the treatment of TPO (n=5). Dot-plot analysis of annexin V/PI staining did not demonstrated that TPO reduced the population of apoptotic cells induced by iron-overload (n=5). Also, incubation with TPO did not decrease the iron-induced caspase-3 expression in these cells (n=5). Our findings suggest that iron-overload induces apoptosis in cardiomyocytes and hepatocytes via mitochondrial/caspase-3 pathways and that TPO might exert a protective effect on iron-overload induced apoptosis via the activation of Akt and Erk1/2 pathways in cardiomyocytes.

RNA Interference Against Bcl-2 mRNA Increases Radiosensitivity of Raji Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4601-4601
Author(s):  
Dongmei He ◽  
Yuan Zhang ◽  
Gexiu Liu
Keyword(s):  
Caspase 3 ◽  
Radiation Treatment ◽  
Annexin V ◽  
Treated Group ◽  
Parp Cleavage ◽  
Control Group ◽  
Raji Cells ◽  
Protein Levels ◽  
Significant Difference ◽  
Induced Apoptosis

Abstract Bcl-2 is the prominent member of a family of proteins responsible for dysregulation of apoptosis, and resistance to chemotherapy. It has been shown that reduction in Bcl-2 protein levels could ultimately induce a lower apoptotic threshold and restore chemosensitivity in a variety of malignancies. Short interfering RNA (siRNA) has been evaluated as an attractive and effective tool for suppressing a target protein by specifically digesting its mRNA. In our lab, we have identified a siRNA targeting against Bcl-2 could effectively down-regulate Bcl-2 protein. In this study, we investigated the effect of gamma radiation combined with the siRNA targeting Bcl-2 on proliferation and apoptosis in B-lymphoma Raji cells. The siRNA was introduced into cells using Oligofectamine transfection. Cells were treated with Bcl-2 siRNA alone or with 2–8 Gy dose of (60)Co gamma rays. Expression of Bcl-2 mRNA and protein was assayed by quantitative reverse transcriptase-polymerase chain reaction and Western blotting analysis. Radiosensitivity was determined by clonogenic cell survival assay. Apoptosis was determined by Giemsa staining, Annexin-V binding assay and flow cytomertry. Furthermore caspase-3 activity and poly (ADP-ribose) polymerase (PARP) cleavage were evaluated. Transfection of Raji cells with 100 nmol/L siRNA targeted against Bcl-2 resulted in reduction of Bcl-2 mRNA by 75% compared with control-siRNA treated group and the vehicle control group(p<0.05). The levels of Bcl-2 protein were significantly reduced by 70% compared with the two control groups (p<0.05). There was significant difference in the radiosensitivity of Raji cells in which Bcl-2 was silenced compared with the cells transfected vehicle or control siRNA.Apoptosis index of the Raji cells treated with Bcl-2 siRNA combined with radiation was significantly increased (p<0.05), compared with either control siRNA / radiation combination or radiation-treatment cells alone, or Bcl-2 siRNA-treatment cells alone.Raji cells treated with Bcl-2 siRNA combined with radiation revealed enhanced caspase-3 and PARP cleavage as compared to Bcl-2 siRNA treated cells alone or only irradiated cells. These findings show that Bcl-2 siRNA synergistically enhances radiation-induced apoptosis through the expression of proteins involved in the programmed cell death.

scholarly journals Anti-Apoptotic Effect of Angelica Polysaccharide (APS) on Cryopreservation of Platelets

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3250-3250
Author(s):  
Mo Yang ◽  
Weiqing Su ◽  
Liuming Yang ◽  
Huimin Kong ◽  
Huiling Wei ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Conflicts Of Interest ◽  
Annexin V ◽  
Control Group ◽  
Apoptotic Effect ◽  
Vitro Effect

Abstract Background: Angelica Polysaccharide (APS) is from the root of Radix Angelicae Sinensis (Danggui). Danggui has been used for centuries to treat blood-deficiency related diseases. The hematopoietic effect of Danggui may be related to its constituent, polysaccharide. The effects of angelica polysaccharide on cryopreservation of platelets and megakaryocytes have not been well studied. This study focused on anti-apoptotic effect of APS and TPO on cryopreservation of platelets and megakaryocytes and provided new methods for prolonging the preservation time of platelets in vitro. Methods: The expression of platelet membrane glycoprotein CD41 and CD61, as well as the platelet apoptotic rate, Caspase 3 expression and mitochondrial membrane potential (MMP) were detected by flow cytometry; the anti-apoptotic mechanism of APS by PI3K /AKT signaling pathway was analyzed by Western blot assay. CFU assays were used to determine the effects of APS on megakaryocytic progenitor cells. Analyses of Annexin V, Caspase-3, and Mitochondrial Membrane Potential were conducted in megakaryocytic cell line M-07e. The effects of APS on cells treated with Ly294002, PI3K inhibitor and the effect of APS on the p-AKT were also studied. Results: The platelets were divided into 4 group: control group (4 ℃ stored platelets), APS group (APS-treated platelets stored at 4 ℃), LY294002 group (LY294002-treated platelets stored at 4 ℃) and LY294002+APS group (LY294002+APS treated platelets stored at 4 ℃). The apoptotic rate of platelets in LY294002 group was obviously increased. Compared with control group, the expression of CD41 and CD61 gradually decreased along with the enhancement of LY294002 concentrations (r=-0.953). The apoptotic rate of platelets in LY294002 group was enhanced significantly (P&lt;0.05). While in LY294002+APS group, the apoptotic rate of platelets was significantly reduced (P&lt;0.05) as compare with LY294002 group, which suggest that APS has an anti-apoptotic effect on the cryopreserved platelets. APS decreased the expression of Caspase-3 and inhibited the reduction of mitochondrial membrane potential induced by LY294002. Moreover, APS increased the activation of PI3K /AKT pathway in Platelets . We further analyzed the in vitro effect of APS on CFU-MK formation. APS (50 ug/ml) enhanced TPO (50 ng/ml) -induced CFU-MK formation (p=0.06, n=4). APS also significantly enhanced PDGF, bFGF and VEGF-induced CFU-MK formation (n=4). Moreover, the anti-apoptotic effect of APS in M-07e cells was also demonstrated by Annexin-V, Caspase-3, and JC-1 assays. Adding LY294002 alone increased the percentage of cells undergoing apoptosis. However, additional of APS to LY294002-treated cells reversed the percentage of cells undergoing apoptosis. Furthermore, addition of APS significantly increased the p-AKT. Conclusion: APS, like TPO, has an anti-apoptotic effect on the cryopreserved platelets and megakaryocytes through activating PI3K/AKT, decreasing the expression of Caspase-3 and inhibiting the reduction of MMP. Disclosures No relevant conflicts of interest to declare.

scholarly journals Protective effect of agmatine against cisplatin-induced apoptosis in an auditory cell line

2020 ◽  
Author(s):  
Euyhyun Park ◽  
Se Hee Lee ◽  
Hoyoung Lee ◽  
Young-Chan Kim ◽  
Hak Hyun Jung ◽  
...  
Keyword(s):  
Protective Effect ◽  
Cell Line ◽  
Mitochondrial Function ◽  
Caspase 3 ◽  
Annexin V ◽  
Oxygen Species ◽  
Protective Effects ◽  
Cellular Apoptosis ◽  
Induced Apoptosis ◽  
Significant Protective Effect

AbstractObjectivesAgmatine, an endogenous metabolite of arginine, is known to have antioxidant activity, protect mitochondrial function, and confer resistance to cellular apoptosis. The aim of this study was to evaluate the protective effects of agmatine against cisplatin-induced cellular apoptosis in an auditory cell line.MethodsHEI-OC1 cells were co-treated with agmatine at different concentrations and 15 µM cisplatin for 48 h. Cell viability was measured and annexin V-FITC/propidium iodide (PI) staining was performed to analyze apoptosis. The levels of intracellular reactive oxygen species (ROS) were measured using flow cytometry. The expression of BAX (Bcl2-associated X protein) and the enzymatic activity of caspase-3 were measured to examine the pathway of apoptosis induction.ResultsCo-treatment with 8 mM agmatine protected HEI-OC1 cells against cisplatin-induced cell apoptosis. Agmatine exerted a significant protective effect against 15 µM cisplatin when applied for 48 h and reduced the proportion of necrotic and late apoptotic cells. Agmatine did not reduce the cisplatin-induced increase in ROS but decreased the expression of BAX and the activity of caspase-3.ConclusionsAgmatine protected against cisplatin-induced cellular apoptosis in an auditory cell line. These effects were mediated by the protection of mitochondrial function and inhibition of apoptosis.

Lobaplatin induces pyroptosis in cervical cancer cells via caspase-3/GSDME pathway

2021 ◽  
Vol 21 ◽  
Author(s):  
Jie Chen ◽  
Lili Ge ◽  
Xiaoyan Shi ◽  
Juan Liu ◽  
Hongjie Ruan ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Flow Cytometry ◽  
Cancer Cells ◽  
Caspase 3 ◽  
Specific Inhibitor ◽  
Annexin V ◽  
Cell Counting ◽  
Control Group ◽  
Double Positive ◽  
Cervical Cancer Cells

Background: Increasing evidence shows that GSDME is involved in tumor chemotherapy. Lobaplatin is an important chemotherapy drug for the treatment of cervical cancer. However, the exact mechanism of lobaplatin in the treatment of cervical cancer remains unclear. Objective: In this study, we study whether GSDME is a new mechanism of lobaplatin in the treatment of cervical cancer. Methods: Cell pyroptosis was measured by Cell Counting Kit-8 and flow cytometry analyses. Western blot analysis was used to check proteins expression. Results: The cell viability was significantly decreased by lobaplatin treatment. Compared with the control group, the percentage of pyroptosis (PI and Annexin-V double-positive cells) increased after lobaplatin treatment. In addition, lobaplatin induced caspase-3 activation and GSDME cleavage. z-DEVD, a specific inhibitor of caspase-3, reduced lobaplatin-mediated GSDME cleavage and concurrently inhibited pyroptosis. More importantly, GSDME deficiency obviously reduced lobaplatin-induced pyroptosis. Conclusion: These data demonstrated that caspase-3/GSDME axis contributed to the lobaplatin-mediated pyroptosis in cervical cancer cells. This finding indicates that GSDME-mediated pyroptosis is a new mechanism for lobaplatin to kill tumor cells and suggest that caspase-3/GSDME pathway offer new insights into cancer chemotherapy.

scholarly journals The Protective Effect of Aspirin Eugenol Ester on Oxidative Stress to PC12 Cells Stimulated with H2O2 through Regulating PI3K/Akt Signal Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Zhen-Dong Zhang ◽  
Ya-Jun Yang ◽  
Xi-Wang Liu ◽  
Zhe Qin ◽  
Shi-Hong Li ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Protective Effect ◽  
Pc12 Cells ◽  
Caspase 3 ◽  
Signal Pathway ◽  
Control Group ◽  
Cell Viability Assay ◽  
Viability Assay ◽  
Induced Apoptosis ◽  
Aspirin Eugenol Ester

Aspirin eugenol ester (AEE) is a new pharmaceutical compound esterified by aspirin and eugenol, which has anti-inflammatory, antioxidant, and other pharmacological activities. This study is aimed at identifying the protective effect of AEE against H2O2-induced apoptosis in rat adrenal pheochromocytoma PC12 cells and the possible mechanisms. The results of cell viability assay showed that AEE could increase the viability of PC12 cells stimulated by H2O2, while AEE alone had no significant effect on the viability of PC12 cells. Compared with the control group, the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were significantly decreased, and the content of malondialdehyde (MDA) was significantly increased in the H2O2 group. By AEE pretreatment, the level of MDA was reduced and the levels of SOD, CAT, and GSH-Px were increased in H2O2-stimulated PC12 cells. In addition, AEE could reduce the apoptosis of PC12 cells induced by H2O2 via reducing superoxide anion, intracellular ROS, and mitochondrial ROS (mtROS) and increasing the levels of mitochondrial membrane potential (ΔΨm). Furthermore, the results of western blotting showed that compared with the control group, the expression of p-PI3K, p-Akt, and Bcl-2 was significantly decreased, while the expression of Caspase-3 and Bax was significantly increased in the H2O2 group. In the AEE group, AEE pretreatment could upregulate the expression of p-PI3K, p-Akt, and Bcl-2 and downregulate the expression of Caspase-3 and Bax in PC12 cells stimulated with H2O2. The silencing of PI3K with shRNA and its inhibitor-LY294002 could abrogate the protective effect of AEE in PC12 cells. Therefore, AEE has a protective effect on H2O2-induced PC12 cells by regulating the PI3K/Akt signal pathway to inhibit oxidative stress.

scholarly journals Platelet and Immune Characteristics of Patients with Immune Thrombocytopaenia Non Responders to Therapeutic Treatments

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 1089-1089
Author(s):  
Elena Monzón Manzano ◽  
Raul Justo Sanz ◽  
Diana Hernández ◽  
Teresa Álvarez Roman ◽  
Ihosvany Fernandez-Bello ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Response ◽  
Sialic Acid ◽  
Research Funding ◽  
Caspase 3 ◽  
Mononuclear Cells ◽  
Ricinus Communis ◽  
The Other ◽  
Control Group ◽  
Platelet Surface

Introduction: Mechanisms leading to diminished platelet counts in immune thrombocytopaenia (ITP) appear to be multifactorial: autoantibodies, autoreactive CD8+ cytotoxic T cells, enhanced apoptosis and loss of sialic acid which mediates platelet clearance through the Ashwell-Morell receptors present in hepatocytes. Differential involvement of each of them might condition the ability of patients with ITP to respond to treatments. We aimed to examine platelet features and the immunological state of patients with ITP who do not respond to any treatment to detect the unique characteristics of this group. Methods: This was an observational, prospective and transversal study. Patients with chronic primary ITP were included: 28 ITP patients without treatment for at least 6 months (UT-ITP); 36 responders to agonists of thrombopoietin receptors (TPO-RA); and 14 ITP patients who did not respond to first- and second-line treatments (NR-ITP). A healthy control group (n=104) was also included in the study. Active caspase-3, -7, -8 or -9 were determined by flow cytometry using CaspaTag kits (Millipore, Madrid, Spain) in PRP diluted with HEPES-buffer containing 2 mM Ca2+ and 2 mM Gly-Pro-Arg-Pro (Sigma-Aldrich, Madrid, Spain) to prevent fibrin formation . Platelet surface glycan exposure was analysed by determining the binding of lectins by flow cytometry. To do so, washed platelets were incubated with 1 μg/ml Alexa fluor 488-conjugated wheat germ agglutinin lectin (WGA, Invitrogen, Spain) or with 1 μg/ml FITC-conjugated Ricinus communis agglutinin (RCA, Vector Labs, UK). WGA binds to sialic acid and N-acetylglucosaminyl residues, and RCA is a galactose-specific legume lectin which binding serves as an indirect measurement of the loss of sialic acid. Peripheral blood mononuclear cells (PBMCs) subsets were analysed by flow cytometry using specific antibodies. Experimental data was analysed using SPSS 9.0 software (SPSS Inc., Chicago, IL). Results: Platelets from TPO-RA treated and from NR-ITP patients had increased caspase-3, -7, -8 and -9 activities (Figure 1A). Platelets from NR-ITP patients exposed less sialic acid and more N-acetylglucosaminyl residues than the other groups (Figure 1B). Binding of WGA and RCA correlated with caspase activities (Table 1). Distribution of lymphocytes, monocytes and natural killer cells is shown in Table 1. NR-ITP patients had an increased proportion of B lymphocyte (LB), maybe due to a significant rise in the fraction of naive LB cells, and a diminution in LTreg subset. Whereas classical monocytes was increased, nonclassical monocyte fraction was decreased in the UT-ITP and NR-ITP groups. NR-ITP patients also presented an increased CD16+CD56bright cells fraction and a diminished NK CD16+CD56dim subset. TPO-RA-treated patients seemed to recover an immune homeostasis similar to healthy controls (monocyte and NK cells subset distribution and LTreg count similar to control group). It is of interest to note the relationship between loss of sialic acid from platelet surface glycans and Tregs count: the most reduced surface exposure of sialic acid, the less Treg count (Figure 2). Conclusions: Platelets from NR-ITP patients had more signs of apoptosis and a different composition of surface glycans, accompanied by a diminished LTreg population, a higher LB naïve percentage, and an increased CD16+CD56bright cells fraction in circulation, indicating a severe deregulation of the immune system. Since an inverse correlation was observed between loss of sialic acid and LTreg count, a potential relationship between glycan composition on the platelet surface and immune response is suggested, positing terminal sugar moieties of the glycan chains as aetiopathogenic agents in ITP. On the other hand, TPO-RA appears to have a beneficial effect on immune response. Nevertheless, one of the limitations of our study was that patients were recruited once the response to TPO-RA was achieved; therefore, a longitudinal study would provide more information regarding TPO-RA effects. This work was supported by grants from the FIS-FONDOS FEDER (PI15/01457, NB). NVB holds a Miguel Servet tenure track grant from FIS-FONDOS FEDER (CP14/00024). Disclosures Álvarez Roman: Roche: Consultancy, Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; Bayer: Consultancy, Speakers Bureau; Novartis: Consultancy, Speakers Bureau; Amgen: Consultancy, Speakers Bureau; Takeda: Research Funding; NovoNordisk: Consultancy, Speakers Bureau; CSL Behring: Consultancy, Speakers Bureau; Sobi: Consultancy, Speakers Bureau. Fernandez-Bello:Novartis, Pfizer, ROCHE, Stago: Speakers Bureau. Martín:SOBI: Research Funding; Novartis, Pfizer, ROCHE, Novo Nordisk: Speakers Bureau. Rivas Pollmar:Novartis, Pfizer, ROCHE, Novo Nordisk: Speakers Bureau; SOBI: Research Funding. Canales:Novartis: Honoraria; Takeda: Speakers Bureau; iQone: Honoraria; Sandoz: Honoraria; Celgene: Honoraria; SOBI: Research Funding; Karyopharm: Honoraria; F. Hoffmann-La Roche Ltd: Honoraria, Speakers Bureau; Gilead: Honoraria; Janssen: Honoraria, Speakers Bureau. Jimenez-Yuste:Bayer, CSL Behring, Grifols, Novo Nordisk, Octapharma, Pfizer, Roche, Sobi, Shire: Consultancy, Honoraria, Other: reimbursement for attending symposia/congresses , Research Funding, Speakers Bureau. Butta:Novartis: Consultancy; Roche, Pfizer: Speakers Bureau.

scholarly journals A Higher Frequency Administration of the Nontoxic Cycloartane-Type Triterpene Argentatin A Improved Its Anti-Tumor Activity

Molecules ◽  
2020 ◽  
Vol 25 (8) ◽  
pp. 1780 ◽  
Author(s):  
Zaira Tavarez-Santamaría ◽  
Nadia J. Jacobo-Herrera ◽  
Leticia Rocha-Zavaleta ◽  
Alejandro Zentella-Dehesa ◽  
Beatriz del Carmen Couder-García ◽  
...  
Keyword(s):  
Waste Product ◽  
Industrial Process ◽  
Annexin V ◽  
Control Group ◽  
Healthy Control ◽  
Hct 116 ◽  
Hct 116 Cells ◽  
Induced Apoptosis

Parthenium argentatum (Gray), commonly known as guayule, has been used to obtain natural rubber since the beginning of the 20th century. Additionally, the so called “resin” is a waste product derived from the industrial process. The cycloartane-type triterpene Argentatin A (AA) is one of the main constituents of the industrial waste resin. In this study we evaluated the AA anticancer activity both in vitro and in vivo in the HCT116 colon cancer cells. The apoptosis promotion of AA was assessed by the annexin V/propidium iodide (PI) assay. The senescence was evaluated for SA-β-galactosidase, and PCNA was used as a marker of proliferation. Its antitumor activity was evaluated using a xenograft mouse model. The results indicated that AA-induced apoptosis in HCT-116 cells and was positively stained for SA-β-galactosidase. In the xenografted mice test, the administration of AA at the dose of 250 mg/kg three times a week for 21 days reduced tumor growth by 78.1%. A comparable tumor reduction was achieved with cisplatin at the dose of 2 mg/kg administered three times a week for 21 days. However, nude mice treated with AA did not lose weight, as they did remarkably when treated with cisplatin. Furthermore, the animals treated with AA showed similar blood profiles as the healthy control group. These data indicate the low toxicity of AA compared to that shown by cisplatin.

Multiple Signal Pathways Involved in Crocetin-Induced Apoptosis in KYSE-150 Cells

Pharmacology ◽  
2019 ◽  
Vol 103 (5-6) ◽  
pp. 263-272 ◽  
Author(s):  
Sheng Li ◽  
Yuhua Qu ◽  
Xiu-Yin Shen ◽  
Ting Ouyang ◽  
Wen-Bin Fu ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Squamous Carcinoma ◽  
Annexin V ◽  
Signal Pathways ◽  
Dependent Manner ◽  
Apoptosis Pathway ◽  
Anticancer Properties ◽  
Multiple Signal ◽  
Underlying Mechanisms ◽  
Induced Apoptosis

Background: Crocetin is a carotenoid extracted from the traditional Chinese medical herb saffron. Previous studies have demonstrated that crocetin possesses anticancer properties that are effective against various cancers. As an extension of our earlier study, the present study explored the underlying mechanisms in crocetin’s anticancer effect on KYSE-150 cells. The phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT), Mitogen-activated protein kinases (MAPK), and p53/p21 signal pathways play an important role in carcinogenesis, progression, and metastasis of carcinoma cells. Thus, we investigated crocetin’s effects on the PI3K/AKT, MAPK, and p53/p21 pathways in esophageal squamous carcinoma cell line KYSE-150 cells. Methods: KYSE-150 cells were treated with various concentrations of crocetin. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltertrazolium bromide assay, Annexin V/PI stain as well as Rh123 stain were used to evaluate the cell viability, apoptosis, and MMP. Western blot was used to detect the expression of PI3K, AKT, ERK1/2, p38, c-Jun NH-terminal kinase (JNK), P53, P21, Bcl-2, Bax, and cleaved caspase-3, which were associated with cell proliferation and apoptosis. Results: Our results showed that crocetin significantly inhibited the proliferation of KYSE-150 cells in a dose- and time-dependent manner. Crocetin also markedly induced cell apoptosis. Furthermore, we have found that crocetin not only inhibited the activation of PI3K/AKT, extracellular signal–regulated kinase-1/2 (ERK1/2), and p38 but also upregulated the p53/p21 level. These regulations ultimately triggered the mitochondrial-mediated apoptosis pathway with an eventual disruption of MMP, increased levels of Bax and cleaved caspase-3, and decreased levels of Bcl-2. Conclusions: These findings suggested that crocetin interfered with multiple signal pathways in KYSE-150 cells. Therefore, this study suggested that crocetin could potentially be used as a therapeutic candidate for the treatment of esophageal cancer.

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