• Archives of Pharmacal Research
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• v.18 no.2
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• pp.90-99
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• 1995

The effect of protein kinase C inhibitors, sturosporine and 1-(5-isoquinolinyl sulfonyl)-2-methyl piperazine(H7) on in vitro differentiation of erythroid progenitor cells which were isolated from spleens of mice infected with the anemia-inducing strain of Friend virus were examined. Erythropoietin-mediated differentitation of erythroid progenitor cells, as determined by the incorporation of $^{59}Fe$ into protoporphyrin, was inhibited by staurosporine and H7 in a concentration -dependent manner. Scatchard analysis of the $^3H-phorbol-12$, 13-dibutyrate binding to erythroid progenitor cells revealed that at the high affinity sites the dissociation constant was 22nM and the maximum number of $^3H-phorbol-12$, 13-dibutyrate binding to erythroid progenitor cells revealed that at the high affinity sites the dissociation constant was 22nM and the maximum number of $^3H-phorbol-12$, 13-dibutyrate binding sites per cell was approximately $3.7\times10^5$. Cytosonic protein kinase C was isolated from erthroid progenitor cells and then purified by sequential column chromatogrphy. Two isoforms of protein kinase C were found. Photoaffinity labeling of the purified protein kinase C samples with $^3H-phorbol-12$12-myristate 13-acetate followed by analysis of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and autofluorography showed radiolabeled 82-KDa pepticles. Rediolabeling of the 82-KDa peptides with $^3H-phorbol-12$myristate 13-acete was almost completely blocked by excess unlabeled phorbol 12-myristate 13-acetate was almost 12-muristate 13-acetate-promoted phosphorylation with the puyrified protein kinase C samples showed that the phosphorylation of 82-KDa peptides was increased as the concentration of phorbol 12-myristate 13-acetate was increased from $10^{-8}M{\;}to{\;}10^{-4}$M. In light of the findings that erythroid progenitor cells possessed an abundance of protein kinase C and that stauroporine and H7 inhibited erythroid differentiation, it seemed likely that protein kinase C would play a role in the erythroid progenitor cell development.

• The Korean Journal of Pharmacology
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• v.31 no.3
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• pp.365-375
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• 1995

The anemia-inducing strain of Friend virus (FVA) is a murine retrovirus which stimulates the proliferation of erythroid progenitor cells. The progenitor cells synthesized by FVA-stimulation are unable to proceed with differentiation and accumulate in the spleen resulting in splenomegaly in infected mice. Using FVA-inoculated mice as a model, we have investigated the antiretroviral effects of 2',3'-dideoxycytidine (ddC) and recombinant $interferon-{\alpha}-A\;(rIFN-{\alpha}-A)$ on FVA infection. The extent of the infection was determined by measuring the weights of the spleens. Daily intraperitoneal injection of ddC (100 mg/kg body weight), $rIFN-{\alpha}-A$ (10 KU/mose) and the combination of both drugs to FVA inoculated mice for 18 days resulted in suppression of the growth of spleens by 15.1%, 52.7% and 61.6%, respectively. When ddC was dissolved in drinking water (0.1 mg/ml) and administered to a group of FVA inoculated mice ad libitum, and $rIFN-{\alpha}-A$ (10 KU/mouse) was intraperitoneally injected daily to another group of ddC (0.1 mg/ml) drinking mice for 18days, the growth of spleens was suppressed by 38.4% and 83.2%, respectively. These results indicate that administration of ddC via drinking water is more effective in suppressing FVA infection than the daily injection of ddC, and that the combined effects ddC and $rIFN-{\alpha}-A$ are not synergistic but additive. In order to determine whether ddC treatment alters the characteristic of the progenitor cells with respect to $Ca^{++}$ uptake, $Ca^{++}$ uptake in erythroid cells and the effect of cyclohexyladenosine (CHA) on the $Ca^{++}$ uptake were studied. $Ca^{++}$ uptake in the erythroid progenitor cells was about 20-fold greater than in mouse erythrocytes and the inhibition of $Ca^{++}$ uptake by CHA was the greatest in the progenitor cells from FVA infected mice which were treated with ddC. The inhibition was obviated by theophylline. Results of CHA binding studies showed that the erythroid progenitor cells contain both high and low affinity CHA binding sites, whereas mose erythrocytes contain only the low affinity CHA binding sites.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3715-3721
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• 2015

Leukemia is a clonal disorder with blocked normal differentiation and cell death of hematopoietic progenitor cells. Traditional modalities with most used radiation and chemotherapy are nonspecific and toxic which cause adverse effects on normal cells. Differentiation inducing therapy forcing malignant cells to undergo terminal differentiation has been proven to be a promising strategy. However, there is still scarce of potent differentiation inducing agents. We show here that Angelica sinensis polysaccharide (ASP), a major active component in Dong quai (Chinese Angelica sinensis), has potential differentiation inducing activity in human chronic erythro-megakaryoblastic leukemia K562 cells. MTT assays and flow cytometric analysis demonstrated that ASP inhibited K562 cell proliferation and arrested the cell cycle at the G0/G1 phase. ASP also triggered K562 cells to undergo erythroid differentiaton as revealed by morphological changes, intensive benzidine staining and hemoglobin colorimetric reaction, as well as increased expression of glycophorin A (GPA) protein. ASP induced redistribution of STAT5 protein from the cytoplasm to the nucleus. Western blotting analysis further identified that ASP markedly sensitized K562 cells to exogenous erythropoietin (EPO) by activating EPO-induced JAK2/STAT5 tyrosine phosphorylation, thus augmenting the EPO-mediated JAK2/STAT5 signaling pathway. On the basis of these findings, we propose that ASP might be developed as a potential candidate for chronic myelogenous leukemia inducing differentiation treatment.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.278-278
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• 1994

HIV와 유사하게 retro virus의 일종인 Friend virus of Anemia strain (FVA)을 BALB/c mice 에게 감염시 reverse transcriptase 의 활성에 의해 비장의 erythroid progenitor cell 에서 증식되므로서 비장의 비대 및 빈혈을 초래하게 된다. 이를 지표로 하여 reverse transcriptase 억제작용을 지닌 항 virus 약물을 천연성분으로 부터 검색하고자 하였다. 우선, 대조약물로 사용한 기존의 항 AIDS 약물인 zidovudine (AZT) 을 FVA 가 감염된 BAB/c mice 에게 18일간 투여시 (약 100 mg/kg/day, p.o. ) 대조군에 비해 비장의 비대 및 reverse transcriptase 활성이 90% 이상 억제되었으며, 이들의 혈청을 정상 BALB/c mice 에게 재투여시에도 유사한 결과를 나타내어 reverse transcriptase 를 억제하므로서 항 virus 작용을 나타낸다는 것을 입증하였다. 그러나, 혈액중 Hemoglobin등 빈혈의 지수는 대조군과 유사한 수치를 나타내므로서 정상으로 회복되지를 못하였다. 이것은 zidovudine 자체가 지니는 골수억제에 의한 부작용 때문인 것으로 고려된다.

• Clinical and Experimental Pediatrics
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• v.46 no.11
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• pp.1139-1142
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• 2003

Human parvovirus(HPV) B19 infection causes erythema infectiosum in children, sometimes red cell aplastic crisis with hemolytic anemia and chronic bone marrow failure in immunocompromised hosts. HPV B19 is directly cytotoxic for erythroid progenitor cells and inhibits erythropoiesis. Infrequently, HPV B19 inhibits hematopoiesis of three cell lineages and causes transient pancytopenia in patients with hemolytic disorders. We report three patients with hereditary spherocytosis who developed transient aplastic crisis. A HPV B19 infection was confirmed by IgM anti-B19 parvovirus titers and characteristic findings of bone marrow examination as the causative agent associated with severe pancytopenia. Three patients recovered spontaneously after a short period of supportive care with red cell transfusions and intravenous immunoglobulin.

• YAKHAK HOEJI
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• v.39 no.6
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• pp.622-630
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• 1995

Inoculation of Friend anemia virus, which was a Kind of retro virus such as HIV, results splenomegaly, anemia, the increase of WBC counts and reverse transcriptase activity in serum. These results were due to the inhibition of the differentiation of erythroid progenitor cell by the FVA at the spleen. Using these as index of antiviral effects, we pursued the establishment of in vivo screening method for the new anti-ADS drugs. Among zidovudine, didanosine and zalcitabine, which were already approved as anti-AIDS drugs, treatment of zidovudine for 18 days in BALB/c mice inoculated with Friend anemia virus resulted the most potent inhibitory effects on the splenomegaly, the increase of WBC counts and reverse transcriptase activity, but did not recovered the anemia due to the tomcity of zidovudhie itself on the bone marrow. The antiviral effects of zidovudine was reduced in case of zidovudine treatment 7 days after Friend anemia virus inoculation. These results suggested that the sooner treatment of zidovudine would be better improved when the virus was inoculated. Human recombinant interferon itself .alpha. did not showed the antiviral activity against Friend anemia virus and did not also affected the antiviral activity of zidovudine. These results suggested that Friend anemia virus would be used as a tool in vivo screening method for the Lobster of reverse transcriptase.

• Biomedical Science Letters
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• v.15 no.2
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• pp.161-166
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• 2009

Human erythropoietin(EPO) is a glycoprotein that enhances red blood cell production by stimulating proliferation and differentiation of erythroid progenitor cells in the bone marrow. Patients with chronic kidney disease(CKD) suffer from anemia caused by reduced production of EPO in the kidney. Recombinant human EPO protein has been used successfully for the treatment of anemia associated with CKD. Recently, attention has been paid to the development of side effect of EPO, pure red cell aplasia(PRCA), in some patients with CKD. PRCA is a rare disorder of erythropoiesis that leads to a severe anemia due to an almost complete cessation of red blood cell production. EPO-related PRCA is caused by the production of EPO-neutralizing antibodies(Abs) that eliminate the biological activity of EPO as well as endogenous EPO in patients undergoing therapy. Since 1988, almost 200 cases worldwide have been reported with Ab-positive PRCA after receiving EPO therapeutics. The underlying mechanisms of the breaking of immune tolerance to self-EPO have been investigated. Modification of formulation, organic compounds of container closures, and route of administration has been suggested for the possible mechanism of increased immunogenicity of EPO. A number of assays have been used to detect Abs specific to EPO. These assays are generally grouped into two major categories: binding Ab assays and neutralizing Ab assays(bioassays). There are several types of binding Ab assays, including radioimmunoprecipitation assay, enzyme-linked immunosorbent assay, and the BIAcore biosensor assay. In vitro cell-based bioassays have been utilized for the detection of neutralizing Abs. Finally, the recent experience with anti-EPO Abs may have considerable implications for the future development and approval of EPO preparations. Also, considering that millions of patients are being treated with EPO, clinicians need to be aware of signs and consequences of this rare but severe clinical case.

• Journal of Ginseng Research
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• v.44 no.2
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• pp.291-299
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• 2020

Background: Ginseng (G) and Ligustrum lucidum Ait (LLA) are core traditional Chinese medicines in treating myelosuppression formula. The present study was designed to profile effect of G and LLA herb pair (G-LLA) on myelosuppressed mice. Methods: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (Cy). Hematopoietic function of bone marrow was measured by hemopoietic progenitor cell culture and peripheral blood count, and serum hemopoietic factors were tested by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. HPLC was used to measure 20 potential chemical components related to myelosuppression, including ginsenoside Rg₁, Re, Rb₁, Rc, Rb₂, Rb₃, Rd, Rk₃, Rh₄, 20 (S)-Rg₃, 20 (R)-Rg₃, Rk₁, Rg₅, salidroside, and so on. Results: G, LLA, and G-LLA improved the amount of peripheral blood cells and bone marrow cells of myelosuppressed mice (P < 0.01). They significantly increased the colony quantity of colony-forming unit-granulocyte macrophage, burst-forming unit-erythroid, colony-forming unit-erythroid, and colony-forming unit-megakaryocyte and amount of G₂/M and S phase cells (P < 0.01). They also significantly decreased the amount of hematopoiesis-related cytokines (P < 0.01). The content of chemical components in G-LLA changed, and the change of rare saponin was the most obvious. Conclusion: These results show that G-LLA herb pair might produce synergistic or complementary compatibility effects on bone marrow suppression after chemotherapy. It suggests that the substance basis of G-LLA for treating bone marrow suppression may be effective chemical components.