[Effects of Shenxiong pill on infarct brain volumes and NF-kappaB expression in SD rats with middle cerebral artery occlusion (MCAO)].

OBJECTIVE: To investigate the effects of Shenxiong Pill on the infarct volume and expression of NF-kappaB in brains of rats with middle cerebral artery occlusion. METHODS: 169 SD rats were randomly divided into five groups: normal group, sham group, model group, cyclophosphamide group and Shenxiong Pill group. MCAO rat models were established by string ligation (for model, cyclophosphamide-treated and Shenxiong-treated groups). Rats in the Shenxiong Pill group was further randomly divided into sub-groups, receiving a range of high dose treatment (5 to 20 times of clinical dosage). Brains of the rats were examined 48 h or 72 h after interventions in a random order. Image processing software was used in the calculation of volume of cerebral infarction. Conventional HE staining was used for observation of brain tissue. Immunohistochemical method was used to determine NF-kappaB expression. RESULTS: Compared with the model group, rats treated with Shenxiong Pill and cyclophosphamide had lower infarct brain volumes (P < 0.05). NF-kappaB positive inflammatory cells were not found in the normal and sham groups. But the MCAO model rats had increased numbers of NF-kappaB positive inflammatory cells and higher integral optical density of NF-kappaB over time. Compared with the model group, lower numbers and expression of NF-kappaB positive inflammatory cells were found in those treated with Shenxiong Pill (P<0. 05). Higher dosage of Shenxiong was associated with lower numbers and expression of NF-gB inflammatory cells (P<0. 05). CONCLUSION: Shenxiong Pill can reduce pathological damage to brains as a result of cerebral ischemia, possibly through inhibiting the expression and activation of NF-kappaB.

Effect comparison of different formulation of Dang-Gui-Bu-Xu-Tang on myelosuppression mouse.

OBJECTIVE: To compare effect of different formulation of Dang-Gui-Bu-Xu-Tang(DGBXT) on myelosuppression mouse. METHODS: HPLC was used to measure active ingredients of two DGBXT formulations. Hemopoesic function of bone marrow was measured by hemopoietic progenitor cell culture and peipheral blood count. And hemopoietic factors in bone marrow were tested by ELISA. RESULTS: The content level of astragaloside A in granule formulation was higher than that in decoction and the content ratio of active ingredient was close to 5 : 1. Two DGBXD formulations could significantly improve amount of peripheral blood cells, and bone marrow cells of bone marrow suppression mice (P<0.05). DGBXD granule formulation significantly increased the colony quantity of all progenitor cell lines and amount of G(2)/M and S phase cells (P<0.05). It also significantly decreased amount of G(0)/G(1) phase cells in the bone marrow and was more effective (P<0.05). CONCLUSIONS: DGBXT decoction and the granule formulation all can improve the hematopoietic function of bone marrow suppression mouse. They can improve quantities of peripheral blood and nucleated bone marrow cells, and yield of the hematopoietic stem/progenitor cells in vitro colony; balance the expression of cytokine (EPO, TPO and GM-CSF) in bone marrow microenvirement. They can also facilitate hematopoietic stem/progenitor cells to enter the cell cycle. And the effect of granule formulation is more satisfactory.

[Effect of tanshinone IIA pretreatment on IL-1ß and RelA mRNA expression in rats with focal cerebral ischemia].

OBJECTIVE: To observe the effect of tanshinone IIA (TS IIA) pretreatment on the expression of the inflammatory factor IL-1ß and RelA mRNA in rats with focal cerebral ischemia. METHODS: A total of 100 adult male SD rats were randomly divided into 6 groups, namely the model, ischemic preconditioning (IPC), TSIIA preconditioning, TSIIA treatment, sham-operated, and blank control groups. In the former 4 groups, rat models of focal cerebral ischemia were established with corresponding treatments. The expressions of IL-1ß and RelA mRNA in each group were detected using RT-PCR. RESULTS: All the groups showed expressions of IL-1ß and RelA mRNA with the exception of the blank control group. Compared to the model group, TSIIA preconditioning group, TSIIA treatment group, and IPC group all had significantly reduced expression of IL-1ß and RelA mRNA (P < 0.05). The expressions were lower in IPC group than in TSIIA preconditioning group and TSIIA treatment group(P < 0.05), and no significant difference was found in the expressions between the latter two groups. CONCLUSION: The protective effect of pretreatment with TS IIA against cerebral ischemia is related to the reduction of IL-1ß and RelA mRNA expressions.

Experimental study of the bone marrow protective effect of a traditional Chinese compound preparation.

This study investigated the effect of Wei Gan Li on bone marrow haemopoiesis in myelosuppressed anaemic mice induced by (60)Cogamma, cyclophosphamide and chloramphenicol. After treatment with Wei Gan Li, colony forming units of granulocyte macrophages, erythroid cells, burst forming unit-erythroid cells, megakaryocytes and the proliferation of bone marrow stromal cells were measured by in vitro cell culture techniques. Bone marrow haemopoietic stem cells were measured by flow cytometry. The peripheral blood cell count was found to have significantly recovered after Wei Gan Li administration and the proliferation of bone marrow haemopoietic stem/progenitor cells and BMSC were also significantly increased. Wei Gan Li was therefore found to promote the recovery of bone marrow haemopoietic function in myelosuppressed anaemic mice.

Effect of nerve growth factor on erythropoiesis in mice and its underlying mechanism.

This study was aimed to explore the effect of nerve growth factor (NGF) on erythropoiesis and to elucidate the underlying mechanism. Using flow cytometry, colony forming assay, blood cell counter, fluorescent real-time quantitation PCR, and enzyme-linked immunosorbent assay (ELISA), the changes in the bone marrow cells (BMCs) proliferation cycle, CFU-E and BFU-E counts, the peripheral blood erythroid related parameters, kidney EPO, BMC GM-CSF, spleen EPO receptor (EPOR) mRNA expression, and serum EPO, GM-CSF, and IL-1 concentrations were all determined after NGF was injected intramuscularly into the thigh of mice, meanwhile the change of BFU-E and CFU-E counts and its relationship with EPO, IL-3 were investigated. The results indicated that the cell proportion in S+G2/M phase, the CFU-E and BFU-E counts of BMCs and the spleen EPOR mRNA expression in injection of NGF (7.5 microg/kg) for 7 days were significantly higher than that in injection of physiological saline for 13-19 days; red blood cell, hemoglobin, and reticulocyte counts increased as well. In vitro, NGF stimulated a dose-dependent increase of CFU-E colonies formation in the semisolid culture system with or without exogenous EPO; the colony counts in the system with NGF alone were significantly higher than that in the system with exogenous EPO alone. The BFU-E counts in the system with exogenous NGF and IL-3 were significantly higher than that in the system with exogenous EPO and IL-3. It is concluded that the NGF promotes the responsibility of hematopoietic cells to EPO and activates the same signal transduction pathway as EPO in hematopoietic cells, and then accelerates the BMCs into mitosis, the HSCs differentiating into erythroid cells, and CFU-E and BFU-E formation.

[The effect of NGF on hematopoiesis of granulocyte-macrophage in normal and radiation, chemistry injury mice].

OBJECTIVE: To explore the effect of nerve growth factor (NGF) on hematopoiesis of granulocyte-macrophage and to elucidate the underlying mechanism. METHODS: Using flow cytometry, colony forming assay, blood cell count, fluorescent real-time quantitation PCR, and enzyme-linked immunosorbent assay (ELISA), changes in the bone marrow cells (BMCs) proliferation cycle, CFU-GM counts, the peripheral white blood cell (WBC)counts, BMC GM-CSF mRNA quantities, and serum GM-CSF, IL-3 concentrations were observed after NGF was injected intramuscularly into the thigh of normal and combined 60Co gamma ray irradiated and cyclophosphamide peritoneal cavity injected(combined radiation and chemistry injury mice, 60Co-Cy injury) mice. The effect of NGF on CFU-GM formation in vitro cooperation with exogenous GM-CSF or alone was also observed. RESULTS: An injection of NGF [7.5 microg/(kg x d) or 10 microg/(kg x d)] lasted 7 days, the S+G2/M proportions and the CFU-GM counts of BMCs rose greater than injection of physiological saline (PS) in normal and 60Co-Cy injury mice; the serum GM-CSF,IL-3 concentration in 60Co-Cy injury mice rose obviously. In vitro, NGF stimulated a dose-dependent increase of CFU-GM colonies formation in the semisolid culture system with exogenous rmGM-CSF. CONCLUSIONS: NGF cooperates with hematopoietic stimulating factors in culture system to promote the CFU-GM colonies in normal and 60Co-Cy injury mice in vitro. NGF accelerates the BMCs into mitosis, the hematopoietic stem cells differentiation into granulocyte-macrophage hematopoietic cells, the CFU-GM formation and promotes the white blood cells count in normal and 60Co-Cy injury mice in vivo, also NGF stimulates the secreting of GM-CSF and IL-3 in 60Co-Cy injury mice,this may be one of the reasons why NGF stimulate the granulocyte-macrophage hematopoiesis repair after combined radiation and chemistry injury.

[Effect of weiganli on level of FL in bone marrow and serum of myelosuppressed anemic mice].

OBJECTIVE: To study the effect of Weiganli on the level of FL in bone marrow and serum of myelosuppressed anemic mice, and to explore it's function on hematopoietic regulation. METHOD: Models of myelosuppressed anemic mice were induced by radiation and chemotherapeutic drug, and the mice were randomly divided into normal group, myelosuppressed anemic group, and Weiganli group (high dose 100 g x L(-1), medium dose 50 g x L(-1), low dose 25 g x L(-1)). Effect of Weiganli on the number of the peripheral blood cells and bone marrow nucleated cells (BMC) were evaluated. Effect of Weiganli on the level of FL (Flt3 ligand) was investigated by ELISA technique. RESULT: High dose of Weiganli could significantly increase granulocytes, erythrocytes, Hb and BMC, while both the medium dose and the low dose had more significant action in increase of platelet. The level of FL in bone marrow and serum were lower in Weiganli group than that in myelosuppressed anemic group, especially in high and medium dose group. CONCLUSION: The myelosuppresion of mice which induced by radiation and chemotherapeutic drug could be significantly relieved by Weiganli.

Effects of weiganli on the hemopoietic function of the myelosuppressed anemic mice.

OBJECTIVE: To study the effect of weiganli ([Chinese characters: see text]) on bone marrow hemopoiesis. METHODS: The effects of weiganli on the peripheral blood picture and the number of bone marrow nucleated cells (BMCs) were observed in myelosuppressed anemic model mice, and the effects of weiganli on the growth of colony forming unit-granulocyte macrophage (CFU-GM), colony forming unit-erythroid (CFU-E), burst forming unit-erythroid (BFU-E), colony forming unit-megkaryocyte (CFU-Meg) were investigated by in vitro cell culture technique. The hemopoietic stem cells (HSCs, c-kit+) in bone marrow were double stained with fluorescent antibody PE-C-Kit and FITC-CD45, and the HSCs (c-kit+) were counted by flow cytometer with CD45/SSC (side scatter) gating. RESULTS: Peripheral blood cell counts and the number of BMCs were significantly improved after weiganli administration; and bone marrow hemopoietic stem/progenitor cells were significantly increased. CONCLUSION: Weiganli can effectively promote the recovery of hemopoietic function in the myelosuppressed anemic mice.

[Effect of the recombinant staphylokinase on pancreatic ischemia in severe acute pancreatitis of rats].

OBJECTIVE: To investigate the changes in plasma endothelin-1 (ET-1) , von Willebrand factor (vWF), serum 6-keto-prostaglandin(1alpha) (PGF(1alpha)) , thromboxane B2 (TXB2), platelet aggregation rate maximum (PAGm) and pancreatic blood flow after reproduction of severe acute pancreatitis (SAP) in rat, and the effect of recombinant staphylokinase (r-Sak) on SAP. METHODS: Eighty-one SD rats were divided randomly into the sham-operated group (n=27), the SAP model group (n=27), and the r-Sak treatment group (n=27). SAP was produced by administration of 5% sodium taurocholate into the pancreatic duct. The abdomen of rats was opened at 6, 12 and 18 hours after reproduction of SAP for determining the pancreatic blood flow. Blood was obtained at 6, 12 and 18 hours after reproduction of SAP for determining the concentration of plasma vWF with enzyme-labeled immunosorbent assay (ELISA). The concentration of plasma ET-1 and serum 6-keto-PGF(1alpha), and TXB2 were detected by radioimmunoassay. The PAGm induced by collagen and eicosanoids was assessed. RESULTS: Pancreatic blood flow in the SAP group appeared to have a decreasing trend at 6,12 and 18 hours after operation and were significantly decreased at all time points after reproduction of the model, compared with those of the sham-operated group (all P<0.05). The PAGm, content of plasma ET-1, vWF, and TXB2 were significantly increased at all time points after reproduction of the model, while 6-keto-PGF(1alpha) was significantly decreased, compared with those of the sham-operated group (all P < 0.05). Compared with SAP model group, PAGm, the content of plasma ET-1, vWF, and serum TXB2 in the r-Sak group were decreased at all time points, however, the content of serum 6-keto-PGF(1alpha) was increased (all P<0.05). CONCLUSION: The r-Sak can improve pancreatic microcirculation and enhance pancreatic blood flow in rats with SAP, and may be beneficial in the treatment of SAP.

Mechanisms by which Astragalus membranaceus injection regulates hematopoiesis in myelosuppressed mice.

The aim of this study was to investigate the mechanism underlying the effects of Astragalus membranaceus injection (AMI) on myelopoiesis in myelosuppressed mice. At 72 h after cyclophosphamide injection (250 mg/kg), the mice were administered AMI (500 mg/kg) intraperitoneally for 6 consecutive days or an equivalent volume of saline as a control. Murine colony-forming unit-fibroblast (CFU-F) formation, production of IL-6 and GM-CSF by bone marrow stromal cells (BMSC), and bcl-2 protein and mRNA expression in BMSC were measured by CFU-F assay, ELISA, immunohistochemistry and in situ hybridization. The results indicated that AMI improved the hematopoietic microenvironment by enhancing the BMSC survival and proliferation of CFU-F, production of IL-6 as well as GM-CSF by BMSC and bcl-2 protein and mRNA expression in BMSC, which promoted myelopoiesis. The data may provide a mechanistic basis for applying this ancient Chinese herb to promote hematopoiesis as an efficacious adjuvant therapy against myelosuppression induced by anti-cancer therapy.

[Effect of weiganli on apoptosis and expression of Bcl-2, bax protein in bone marrow nucleated cells of anemic mice].

OBJECTIVE: To study the effect of weiganli on apoptosis and expression of Bcl-2 Bax protein in bone marrow nucleated cells ( BMNCs) of anemic mice and investigate its mechanism, so as to find its new plinic use. METHODS: Effects of WeiGanLi on the amount of the peripheral blood cells and BMNCs were determined on the model of myelosuppressed anemic mice. The cell cycle of BMNCs was analyzed by flow cytometer. The apoptosis of BMNCs was detected by TUNEL, and the expression of Bcl-2 and Bax protein were detected by immunohistochemical method. RESULTS: Weiganli could significantly increase the number of RBC, WBC, HGb, Phit as well as the number of BNMCs, significantly active BNMCs from stasis into cell cycling and Markedly decrease the apoptosis of BNMCs after 2.0 Gy 60Co gamma irradition while the expression of Bcl-2 increased and Bax decreased. CONCLUSION: Weiganli can significantly inhibit the apoptosis of BMNCs probably due to its enhancing effects on Bcl-2 expression, and inhibitory effect on Bax expression. In this way, it prevents the blood cells from injury, and contributes to the rehabilitation of the organic blood production.

[Effects of salidroside on bone marrow matrix metalloproteinases of bone marrow depressed anemic mice].

To examine the effect of salidroside on the expression and activities of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in bone marrow (BM) of BM depressed anemic mice by immunohistochemistry and gelatin zymography respectively, and to explore its roles in hematopoietic regulation. Immunohistochemistry showed that the expression of MMP-2 and MMP-9 of bone marrow cells (BMCs) was found in each group. Compared with control group, the expression of MMP-2 and MMP-9 was obviously increased in the model group, low-dose, middle-dose and high-dose salidroside. At day 4 after treatment of radiation and chemotherapy, the peak of the expression of MMP-2 and MMP-9 was found in middle-dose salidroside . At day 8 after treatment of radiation and chemotherapy, the peak of the expression of MMP-2 and MMP-9 was found in low-dose and middle-dose salidroside respectively. Gelatin zymography revealed that 66 kD proMMP-2, 62 kD MMP-2, 86 kD MMP-9 and 94 kD proMMP-9 were detected in control group, and the activity of MMP-9 was stronger among them. After treatment of radiation and chemotherapy, the activity of gelatinases of hemopoietic microenviroment (HM) was obviously decreased, but low-dose, middle-dose and high-dose salidroside could significantly increase the activities of proMMP-9 and MMP-9, attenuate the activity of proMMP-2. These results suggest that salidroside could promote the recovery of hematopoietic function of BM depressed anemic mice by increasing the expression and activity of MMPs, releasing the cytokines from ECM or cell membrane, repairing impaired microvessels of HM and promotion proliferation, migration and differentiation of HSCs.

[Effect of salidroside on bone marrow cell cycle and expression of apoptosis-related proteins in bone marrow cells of bone marrow depressed anemia mice].

OBJECTIVE: To examine the effect of salidroside on bone marrow cell cycle and expression of apoptosis-related proteins in bone marrow cells (BMCs) of bone marrow depressed anemia mice, and to explore its mechanism for hematopoietic regulation. METHODS: The effect of salidroside on peripheral blood cells, BMCs, and bone marrow cell cycle in bone marrow depressed anemia mice was detected by automatic blood cell analysator, white blood count and flow cytometry (FCM)respectively,and the expression of Bcl-2 and Bax of BMCs was detected by immunohistochemistry method simultaneously. RESULTS: It was found that low-dose and high-dose salidroside obviously elevated white blood cells and BMCs, that low-dose salidroside significantly increased platelets and promoted G0/G1-S phase and S-G2/M phase transition of BMCs, that high-dose salidroside markedly promoted S-G2/M phase transition of BMCs, and that both low-dose and high-dose salidroside obviously elevated the proliferation index and the ratio of G2/M phase cells. Additionally, the expression of Bcl-2 in BMCs was increased in low-dose and high-dose salidroside groups, especially the increase was significant in the low-dose salidroside group; moreover, the expression of Bax in BMCs was reduced significantly in both low-dose and high-dose salidroside groups. CONCLUSION: These data suggest that salidroside may promote the recovery of hematopoietic function of the bone marrow depressed anemia in mice by ending off G0/G1-phase arrest, accelerating G0/G1-S phase and S-G2/M phase transition, up-regulating Bcl-2 expression, down-regulating Bax expression, and inhibiting BMCs apoptosis.

[Study on effect of Astragalus membranaceus injection on hematopoiesis in anemic mice with myelosuppression].

OBJECTIVE: To investigate the effect of AMI on hematopoiesis in anemic mice and explore the possible mechanisms. METHODS: Anemic mice model resulted from myelosuppression by irradiation and cytotoxic chemotherapeutic compounds were randomly divided into three groups: treated group I, treated group II and anemic control group. Intraperitoneal doses of AMI(500 mg/kg, 1000 mg/kg) were given to the treated group, and equal doses of physiological saline were given to the anemic control group. On 7 days after treatment, the count of whole blood cells and bone marrow cells were determined by blood auto-analyzer. The numbers of CFU-GM (granulocyte and macrophage colony forming unit), CFU-E (colony forming unit-erythroid), BFU-E (burst forming unit-erythroid), CFU-Meg(colony forming unit-megakaryocyte) were determined by using technique of hematopoietic progenitor cells culture in vitro. Expression of anti-apoptosis protein BcL-XL and BcL-2 in BMC were determined by immunohistochemistry. RESULTS: RBC, HB, PLT (P < 0.05) and BMC (P < 0.01) in treated group I were significantly higher than that of anemic control. The number of CFU-E, BFU-E and CFU-Meg as well as expression of anti-apoptosis protein BcL-XL of BMC in treated group I also were significantly higher than that of anemic control (P <0. 05 or P <0. 01), while numbers of CFU-E as well as expression of anti-apoptosis protenin BcL-XL of BMC in treated group II were higher than that of anemic control (P < 0.05). CONCLUSION: AMI can lesson apoptosis of bone marrow cells and promote hematopoietic progenitor cells to differentiate along the erythroid and megakaryocytoid line by up-regulating expression of antiapoptosis protein BcL-XL of BMC.

[Experimental study on preventive effect of Radix Paeoniae Rubra to restenosis after carotid balloon injury in high fat-diet rabbits].

OBJECTIVE: To observe the preventive effect of Radix Paeoniae Rubra (RPR) to restenosis after carotid balloon injury in rabbits. METHODS: The rabbit model of carotid balloon injury was established adopting Clowes method, and treated with extract of RPR. Component of new genesic intima and expression of proliferating cell nuclear antigen (PCNA) and macrophage was determined by immunochemical stain. The collagen of type I was detected by special staining for blood vessels and the area of new genesic intima was measured by image assay system. RESULTS: RPR could remarkably decreased the PCNA positive expression and inhibit the proliferation of collagen type I and reduce the generating of new intima. CONCLUSION: RPR has significant preventive effect on the restenosis after carotid ballon injury in high fat-diet induced atherosclerotic rabbits.

[Experimental research of PDE5 activity in corpora cavernosa of ganyu qizhi rat model].

OBJECTIVES: To study the PDE5 activity in corpora cavernosa of the Ganyu Qizhi model penis and the effect of the Chinese herbal medicine Shugan Liqi Huoxue (SLH) ointment on it. METHODS: Non-injury stress stimulus method similar to human spirit stress was used to extablish the Ganyu Qizhi animal(rat) model, and the PDE5 activity in corpora cavernosa of the rat penis was measured by the method of immunohistochemistry and computer image analysis. RESULTS: The PDE5 activity in corpora cavernosa of the high-dosage SLH group was significantly different from that of the model group (P < 0.01). CONCLUSION: Ganyu Qizhi may increase the PDE5 activity in corpora cavernosa of the penis while SLH can reduce such activity.