Effects of Danggui and Its Component Ferulic Acid On Hematopoiesis and Platelet Production.

Abstract Abstract 3509 Poster Board III-446 Chinese herbs have been used for the treatment of myelosuppression and thrombocytopenia for centuries and some herb mixtures have been shown to have favorable effects in cancer patients with myelosuppression. Our recent publication has shown that Danggui Buxue Tong, a traditional Chinese medicine decoction containing Radix Angelicae Sinensis (Danggui) and Radix Astragali (Huangqi), promotes hematopoiesis and thrombopoiesis in mouse model (Yang et al, J Ethnopharm, 2009). However, reports about the pharmacological effects of each herb ingredient have been scanty. The root of Danggui is an important ingredient of many commonly used prescriptions in Chinese herbs for promoting blood production. The chemical constituents and the pharmacological action of Danggui have been investigated using modern scientific methods at the beginning of this century. The objective of this study was to compare the effect of Danggui and TPO on hematopoiesis and platelet production in radiation-induced hematocytopenic mouse model. Danggui extracts (10 mg/kg/day) and TPO (10 ug/kg/day) were given by injection daily for 21 days starting from the day after radiotherapy. Peripheral blood platelet, WBC and RBC from Danggui, TPO and water control groups were analyzed at different time-points. On day 21, the mouse was sacrificed and bone marrow cells were harvested for CFU-MK and CFU-GM assays using the plasma clot and methylcellulose methods. The results showed that both Danggui and TPO significantly enhanced the recovery of platelet and WBC count, as well as CFU-MK and CFU-GM formation (n=6). Morphological examination of bone marrow also showed that Danggui treatment significantly increased the recovery of hematopoietic progenitor cells and megakaryocytic cell series. In addition, Danggui also increased the recovery of the body weight and organ weight (liver and spleen) in mouse model. We further analyzed the in vitro effect of Ferulic acid (major component of Danggui) and Astragaloside IV (major component of Huangqi) on hematopoiesis using CFU assay. Ferulic acid plus Astragaloside IV significantly promoted mouse CFU-GEMM formation (n=4). Ferulic acid together with Astragaloside IV also significantly reduced serum free-induced apoptosis in K562 cells by annexin V assay using flow cytometry (n=3). This anti-apoptotic effect can be mediated by reducing the activation of Caspase-3 pathway. In summary, our studies showed that Danggui extracts as well as TPO enhances hematopoiesis and thrombopoiesis in a radiation-induced hematocytopenic mouse model. Ferulic acid, in combination with Astragaloside IV, also has promoting effect on hematopoiesis and anti-apoptotic effect on hematopoietic cells. Disclosures: No relevant conflicts of interest to declare.

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Redox Metal Chelation Ameliorates Radiation-Induced Bone Marrow Toxicity in a Mouse Model

Radiation Research ◽

10.1667/0033-7587(2001)156[0205:rmcari]2.0.co;2 ◽

2001 ◽

Vol 156 (2) ◽

pp. 205-209 ◽

Cited By ~ 2

Author(s):

Rafael M. Nagler ◽

Yoav Eichen ◽

Arnon Nagler

Keyword(s):

Bone Marrow ◽

Mouse Model ◽

Bone Marrow Toxicity ◽

Metal Chelation ◽

Radiation Induced

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Adipose-Derived Mesenchymal Stem Cells (ADSCs) with the Potential to Ameliorate Platelet Recovery, Enhance Megakaryopoiesis, and Inhibit Apoptosis of Bone Marrow Cells in a Mouse Model of Radiation-Induced Thrombocytopenia

Cell Transplantation ◽

10.3727/096368915x688155 ◽

2016 ◽

Vol 25 (2) ◽

pp. 261-273 ◽

Cited By ~ 4

Author(s):

Jiamin Zhang ◽

Shiyuan Zhou ◽

Yi Zhou ◽

Feier Feng ◽

Qianming Wang ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Mouse Model ◽

Bone Marrow Cells ◽

Platelet Recovery ◽

Radiation Induced ◽

Marrow Cells

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The Effect of Polygonum Multiflorum Extracts on Hematopoiesis and Platelet Production.

Blood ◽

10.1182/blood.v106.11.2159.2159 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2159-2159

Author(s):

Mo Yang ◽

Wei Zhe Huang ◽

Nga Hin Pong ◽

Ai Guo Liu Liu ◽

Chi Kong Li ◽

...

Keyword(s):

Bone Marrow ◽

Monoamine Oxidase ◽

Stromal Cells ◽

Blood Cells ◽

Bone Marrow Stromal Cells ◽

Marrow Stromal Cells ◽

Polygonum Multiflorum ◽

Platelet Production ◽

Vitro Effect

Abstract To date, there is no ideal treatment for thrombocytopenia. We have proposed a possible mechanism of serotonin (5-HT) on megakaryocyte (MK) differentiation and platelet formation (Yang et al, Blood, 2003 suppl). The root of polygonum multiflorum thunb (Heshouwu) is an important ingredient of many commonly used prescriptions in Chinese medicine for promoting blood production. Polygonum multiflorum extracts (PME) also inhibit monoamine oxidase (MAO) and increase the level of 5-HT. Therefore, we hypothesize that polygonum multiflorum may have a promoting effect on thrombopoiesis via inhibition of monoamine oxidase (MAO) to increase 5-HT levels. The objective of this study was to investigate the hematopoietic role of PME in irradiated mice. PME (125 mg/kg/day) and TPO (12.5 ug/kg/day) were given by intra-peritoneal injection daily for 21 days starting from the day after irradiation (4 Gy). Peripheral blood platelets, white blood cells (WBC), and red blood cells (RBC) were analyzed from PME, TPO, and vehicle control groups on day 0, 7, 14 and 21. On day 21, the mice were sacrificed and bone marrow cells were harvested for CFU-MK, CFU-GM, BFU-E, CFU-GEMM and CFU-F (fibroblastoid) assays (n=8). We also investigated the in vitro effect of PME on CFU-F formation. Our results showed that PME enhanced the recovery of platelets, WBC, and RBC counts. Moreover, PME also promoted CFU-MK (30 ± 8 vs 15 ± 3 colonies/2 x 105 cells, p<0.01), CFU-GM (38 ± 7 vs 28 ± 7 colonies/2 x 105 cells, p<0.05), BFU-E (19 ± 3 vs 12 ± 4 colonies/2 x 105 cells, p<0.05), and CFU-F formation (36 ± 11 vs 23 ± 7 colonies/2 x 106 cells, p<0.01). Similar results were obtained in TPO-treated group. In in-vitro study, we further analyzed the effect of PME (0–500 ug/ml) on mouse CFU-F formation. The results showed that PME at 100–500 ug/ml significantly enhanced CFU-F formation (p<0.05, n=6). Our studies showed that PME enhances thrombopoiesis in vivo and the growth of bone marrow stromal cells in vitro. Therefore, we speculate that the thrombopoietic activity of PME may be mediated via promoting the bone marrow stromal cells. Although TPO has been effective as an agent for the recovery of platelet production after the onset of thrombocytopenia, long-term clinical usage of TPO may induce potential side effects such as thrombosis. Here we reported that the effect of PME is comparable with that of TPO on hematopoiesis and the production of platelets.

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Recombinant PDGF-BB Enhances Platelet Recovery in Mice with Radiation Induced Thromobocytopenia.

Blood ◽

10.1182/blood.v112.11.1232.1232 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1232-1232

Author(s):

Jie Yu Ye ◽

Godfrey ChiFung Chan ◽

Shing Chan ◽

Jue Cong Luo ◽

Mo Yang

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Imatinib Mesylate ◽

Pdgf Receptor ◽

Hematopoietic Stem ◽

Platelet Production ◽

Apoptotic Effect ◽

Radiation Induced

Abstract Platelet-derived growth factor (PDGF) involves in the regulation of hematopoietic stem cells. Imatinib mesylate (Gleevec), a PDGF receptor inhibitor, induces thrombocytopenia in 50% of patients with chronic myeloid leukaemia (CML). Our previous studies showed that PDGF enhances megakaryocytopoiesis in vitro via its receptors directly. However, the in vivo effect of recombinant PDGF in thrombocytopenic models has not been reported. In this study, we investigated the in vivo effect of PDGF as well as thrombopoietin (TPO) on hematopoietic stem cells and platelet production in a radiation treated mouse model. We also explored its potential molecular mechanisms on thrombopoiesis. A radiation induced myelosuppression with thrombocytopenic model was established using 4-Gy-irradiated mice. PDGF-BB (1 mg/kg/day) and TPO (1 mg/kg/day) were injected separately after radiotherapy into the mice. Peripheral blood platelets, WBC and RBC from PDGF-BB, TPO and control groups were collected and analyzed on day 0, 7, 14 and 21 respectively. The mice were sacrificed on day 21 and their bone marrows were harvested for CFU assays and histology analysis. PDGF-BB, like TPO, showed a significant promoting effect in platelet production (n=5 p= 0.0078) as well as CFU-MK formation (n=5 p=0.0019) compared with the untreated control group. Histology images also indicated that PDGF-BB increased the number of the megakaryocytic cells and its progenitors in bone marrow of irradiated mice. In order to find out the underlying mechanism, we further studied the in vitro effect of PDGF using megakaryocytic cell line M-07e. Our results demonstrated that PDGF alone (100–200ng/ml) significantly activated the PI-3K/AKT signaling pathway (n=9 p=0.014). PDGF plus PI-3K inhibitor (wortmannin, 100nM) was shown to attenuate the expression of AKT (n=9, p=0.0112). Consistently, imatinib mesylate (Gleevec, 1uM) negatively interfered the expression of AKT when combined with PDGF-BB ((n=4, p= 0.0288). PDGF-BB was found to have a similar anti-apoptotic effect as TPO on M07e cells as showed by Annexin V (n=8, p=0.0281), Mitochondrial Membrane Potential (JC-1) (n=7, p=0.0042), and Caspase-3 (n=5, p=0.0080) assays respectively. In summary, our findings showed that PDGF-BB had an in vivo thromobopoietic effect on the radiation induced myelosuppressive mice models. This effect was likely mediated via PDGF receptor with subsequent activation of PI-3K/AKT pathway, which leads to anti-apoptotic effect on megakaryocytes.

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A Novel p53-Based Strategy for Protection of Bone Marrow and Gastrointestinal (GI) Track From Total Body Irradiation (TBI)-Induced Damage.

Blood ◽

10.1182/blood.v116.21.1454.1454 ◽

2010 ◽

Vol 116 (21) ◽

pp. 1454-1454

Author(s):

Chul S. Ha ◽

Suthakar Ganapathy ◽

Zhi Min Yuan

Keyword(s):

Bone Marrow ◽

Body Weight ◽

Mouse Model ◽

Lung Carcinoma ◽

Low Dose ◽

Tumor Volume ◽

Carcinoma Cells ◽

Mouse Xenograft Model ◽

Radiation Induced ◽

P53 Activation

Abstract Abstract 1454 Purpose: Recognizing p53 activation as the primary mechanism that underlies radiation-induced damage to bone marrow and GI track, we aim to explore the use of low dose arsenic-induced transient p53 inhibition as a novel strategy for radiotherapy protection. Methods: Expanding from our recent finding that low-dose of arsenic can suppress radiation-induced p53 activation and subsequent apoptosis, we used a mouse model to test the potential of arsenic in protection of bone marrow and GI track against TBI induced damage. Arsenic pretreatment was carried out by feeding sex-matched mice (4-6 weeks of age) with water with or without 1.0 mg/L sodium arsenic for three days. Mice were randomized into following groups; control; arsenic only; irradiation only; arsenic and irradiation. Mice were then irradiated with 2Gy TBI daily for 7 days. Radiation-induced bone marrow and GI track damages were analyzed histologically with H&E staining 4 weeks after last treatment. Animal body weight was monitored as a measure of toxicity. To test the prediction that arsenic would not provide protection for malignant cells because of their defect in p53, a lung carcinoma cell line NCI-H358 was used to generate a mouse xenograft model. Treatments were initiated three weeks after implantation when the lung carcinoma cells developed into readily visible tumor with an average volume of approximately 100 mm3. Tumor volumes were measured periodically. Tumor volume was calculated using the equation: (volume = length × width × depth × 0.5236 mm3). Two independent experiments were done and the tumor volumes are means ± SE from total of 10 mice per group. Results: Consistent with published results, mice treated with irradiation significantly lost their body weight. TBI-induced body weight loss was effectively prevented by arsenic pretreatment (Fig. 1A). In line with the data of body weight change, radiation treatments were associated with severe damages to small intestine and bone marrow cells, and remarkably, such damages were significantly reduced by low-dose arsenic pretreatment (Fig. 1B). The results together demonstrate the potential of low-dose arsenic to effectively protect bone marrow and GI track to radiation-induced damage. In tumor xenograft models, the tumor volume of the control group continued to increase with time. Arsenic treatment did not have any detectable effect on growth of the implanted tumors. As expected, irradiation with 2Gy of TBI daily for 7 days resulted in marked tumor regression. Arsenic pretreatment showed little effect on irradiation-induced tumor suppression (Fig. 2), indicating that low-dose arsenic pretreatment does not detectably affect the efficacy of radiation, at least in the human lung carcinoma xenograft mouse model. Collectively, the results demonstrate that a brief treatment with low-dose arsenic is associated with a marked protection of bone marrow and GI track without compromising the ability of irradiation to kill carcinoma cells. Conclusion: The animal studies have provided proof-of-principle for the use of low-dose arsenic in radiotherapy protection of bone marrow and GI track, serving as strong rationales to initiate clinical trials. Disclosures: No relevant conflicts of interest to declare.

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Multiplacenta derived stem cell/cytokine treatment increases survival time in a mouse model with radiation-induced bone marrow damage

Cytotechnology ◽

10.1007/s10616-016-9993-y ◽

2016 ◽

Vol 68 (6) ◽

pp. 2677-2686

Author(s):

Jun Li ◽

Yunfang Wei ◽

Lei Yan ◽

Rui Wang ◽

Ying Zhang ◽

...

Keyword(s):

Bone Marrow ◽

Stem Cell ◽

Mouse Model ◽

Survival Time ◽

Cytokine Treatment ◽

Bone Marrow Damage ◽

Radiation Induced

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Comparison of Notoginsenoside R1 and TPO on Thrombopoiesis in Irradiated Mice.

Blood ◽

10.1182/blood.v104.11.3023.3023 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3023-3023

Author(s):

Mo Yang ◽

T.L. Lam ◽

Karen Kwai Har Li ◽

L.K. Leung ◽

KM Lau ◽

...

Keyword(s):

Bone Marrow ◽

Stromal Cells ◽

Blood Cells ◽

Bone Marrow Stromal Cells ◽

Blood Platelets ◽

Marrow Stromal Cells ◽

Chinese Herbs ◽

Promoting Effect ◽

Platelet Production

Abstract The use of combination of Chinese herbs as a treatment for thrombocytopenia has been reported to be effective and safe. We have reported that Angelica Polysaccharide (extracts of Angelica Sinensis) has promoting effect on blood stem cells and megakaryocytopoiesis (Yang et al, Blood, 2002, 100 (11); 53a). Sanqi, Radix Notoginseng, is the dried roots of Panax notoginseng (Burk.) F. H. Chen (Araliaceae). It has been used for treatment of trauma and bleeding due to internal and external injuries. Its main constituents are ginsenosides (a kind of saponins), as well as notoginsenosides (only rich in Notoginseng species). Although Sanqi is a well-known haemostatic drug, its effects and mechanisms on megakaryocyte/platelet production have not been studied. The objective of this study was to compare the effect of a purified notoginsenoside R1 (NR1) and thrombopoietin (TPO) on thrombopoiesis in irradiated mice. NR1 (2.5 mg) and TPO (0.25 ug) were dissolved in distilled water and given by intra-peritoneal injection daily for 14 days starting from the day after radiotherapy. Peripheral blood platelets, white blood cells (WBC), and red blood cells (RBC) were analyzed from NR1, TPO, and vehicle control groups on day 0, 7 and 14. On day 14, the mice were sacrificed and bone marrow cells were harvested for CFU-MK, CFU-GM, BFU-E and CFU-F (fibroblastoid) assays (n=5). Our results showed that NR1 enhanced the recovery of platelets, WBC, and RBC count. Moreover, NR1 also promoted the CFU-F (12 ± 0.7 vs 19 ± 0.38 colonies/2 x 106 cells, p=0.0034), CFU-MK (22 ± 1.9 vs 26 ± 3.8 colonies/2 x 105 cells, p=0.025), CFU-GM (26 ± 5.2 vs 37 ± 4.3 colonies/2 x 105 cells, p=0.002), and BFU-E (13 ± 2.9 vs 18 ± 1.9 colonies/2 x 105 cells, p=0.003) formation. Similar results were obtained in TPO-treated group. In in-vitro study, we further analyzed the effect of NR1 (0–50mM) on mouse CFU-MK formation using a plasma clot colony assay. The results showed that NR1 (20 mM) enhanced TPO (50 ng/ml)-induced CFU-MK formation (19 ± 2.2 vs 30 ± 6.8 colonies/2 x 105 cells, p=0.02, n=5). Furthermore, the effect of NR1 (5–50 mM) on the growth of bone marrow stromal cells was also investigated using CFU-F assay. NR1 (50mM) had a promoting effect on CFU-F growth (18 ± 3.7 vs 24 ± 1.8 colonies/2 x 106 cells, p=0.043, n=5). Our studies showed that NR1 enhances thrombopoiesis in vivo and the growth of bone marrow stromal cells as well as megakaryocytes in vitro. Therefore, we speculate that the thrombopoietic activity of NR1 may be mediated via promoting the progenitor of platelet, megakaryocytes, and bone marrow stromal cells. Although TPO has been used as an agent for the recovery of platelet production after the onset of thrombocytopenia, long-term clinical usage of TPO may induce potential side effects such as thrombosis. Here we reported that the effect of NR1 is comparable with TPO on the production of platelets in irradiated mice.

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Angelica Polysaccharides Promotes Thrombopoiesis through the Phosphatidylinositol 3-Kinase/Akt Pathway.

Blood ◽

10.1182/blood.v112.11.1240.1240 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1240-1240

Author(s):

Mo Yang ◽

Godfrey Chifung Chan ◽

Jie Yu Ye ◽

Chang Liu

Keyword(s):

Bone Marrow ◽

Mouse Model ◽

Progenitor Cells ◽

Caspase 3 ◽

Annexin V ◽

The Body ◽

Akt Pathway ◽

Hematopoietic Stem ◽

Vitro Effect ◽

Phosphatidylinositol 3

Abstract Angelica Polysaccharide is from the root of Radix Angelicae Sinensis (Chinese 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. However, the effects of angelica polysaccharide on hematopoietic stem/progenitor cells and megakaryocyte/platelet lineage have not been well studied. In this study, we specifically investigate the thrombopoietic effect of Angelica polysaccharide (APS) in a mouse model and its molecular mechanism. A myelosuppression mouse model (4-Gy-irradiated) was treated with APS (10 mg/kg/day) and thrombopoietin (TPO) (1 mg/kg/day). Peripheral blood cells from APS, TPO and vehicle-treated samples were counted on days 0, 7, 14 and 21. Then CFU assays were used to determine the effects of APS on the megakaryocytic progenitor cells and other lineages. 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, a Phosphatidylinositol 3-Kinse inhibitor and the effect of APS on the phosphorylation of AKT were also studied. APS as well as TPO significantly enhanced the recovery of platelet and WBC count, and bone marrow CFU-MK and CFU-GM formation (n=6). Morphological examination of bone marrows showed that APS treatment significantly increased the recovery of the megakaryocytic series. APS also increased the recovery of the body weight and organ weight (liver and spleen). However, APS didn’t have a up-regulating effect on TPO production by a ELISA assay. We further analyzed the in vitro effect of APS on CFU-MK and CFU-GM formation. APS (50 ug/ml) enhanced TPO (50 ng/ml) -induced CFU-MK (p=0.06, n=4), and CFU-GM formation (p=0.032, n=6). However, APS alone (00 ug/ml) did not show significant effect on CFU-MK and CFU-GM proliferation (n=6). The effect of APS (1–500 ug/ml) on the growth of bone marrow stromal cells was further investigated using CFU-F (fibroblastoid) assay. APS (50 ug/ml) alone showed a promoting effect on CFU-F growth (p=0.049, n=3). APS (50 ug/ml) also significantly enhanced PDGF, bFGF and VEGF-induced CFU-F formation (n=4). Moreover, the anti-apoptotic effect of APS in M-07e cells was also demonstrated by using Annexin-V, Caspase-3, and JC-1 assays. Addition of 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 phosphorylation of AKT. APS promotes thrombopoiesis in a mouse model. This effect is likely to be mediated by the PI3K/AKT pathway.

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