ABSTRACT
AIM: To investigate the regulatory mechanism of Angelica polysaccharide on hepatic endoplasmic reticulum stress in diabetic KK-Ay mice. MEHTODS: Forty diabetic KK-Ay mice were randomly divided into model group, metformin group, and angelica polysaccharide high, medium, and low dose groups, with 8 mice in each group. 8 C57BL/6J mice were used as blank control group. The mice were gavaged with 400 mg/kg, 200 mg/ kg and 100 mg/kg of angelica polysaccharide in the high, medium and low dose groups, respectively, and 200 mg/kg of metformin hydrochloride in the metformin group, while the normal and model groups were gavaged with equal volume of saline, and fasting blood glucose and body weight were measured weekly. After 4 weeks of gavage, triglyceride (TG), cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels were measured in mice serum; RT-PCR was performed to observe the expression of glucose-regulated protein 78 (GRP78), phosphorylated pancreatic endoplasmic reticulum kinase (p-PERK) and phosphorylated α-subunit eukaryotic initiation factor 2 (p-Eif2α) in liver tissues. mRNA expression; Western blot, immunohistochemistry to detect the protein expression of GRP78, p-PERK, p-Eif2α in mouse liver tissues. HE staining: to observe the histopathological changes in the liver. RESULT: Compared with the blank group, the levels of TC, TG and LDL-C were significantly increased (P<0.01) and the levels of HDL-C were significantly decreased (P<0.01) in the model group; compared with the model group, the levels of TC, TG and LDL-C were significantly decreased (P <0.05, P<0.01) and the levels of HDL-C were significantly increased in the metformin group, angelica polysaccharide high and medium dose groups. Compared with the blank group, the expression of GRP78, p-PERK and p-Eif2α in the model group was significantly upregulated (P<0.01), and the expression of GRP78, p-PERK and p-Eif2α in the angelica polysaccharide high, medium and low dose groups was significantly downregulated (P<0.05, P<0.01), and the high dose group had the best effect compared with the model group. Compared with the model group, the mice in the angelica polysaccharide group showed dense liver tissue, reduced vacuole-like degeneration, reduced liver steatosis, gradually aligned hepatocytes, and clear hepatic sinusoidal structure, and the effect was dose-dependent. CONCLUSION: Angelica polysaccharide significantly improved liver injury in diabetic KK-Ay mice, and its mechanism of action may be related to the inhibition of endoplasmic reticulum stress-related proteins and factors GRP78, p-PERK and p-Eif2α expression by Angelica polysaccharide and improvement of endoplasmic reticulum stress.
ABSTRACT
BACKGROUND: Bone marrow failure is an important pathogenesis of aplastic anemia, and how to reverse bone marrow failure is an issue of concern. OBJECTIVE: To investigate the regulatory effect of angelica polysaccharide on the mitochondrial dysfunction in murine aplastic anemia. METHODS: Seventy-two ICR mice were randomly divided into control, model and treatment groups (n=24 per group). The mice in the latter two groups were used to establish the aplastic anemia models by60Co γ radiation, intraperitoneal injection of cyclophosphamide and intragastric administration of chloramphenicol, and were then given normal saline or 200 mg/kg angelica polysaccharide for 2 weeks, respectively. The control mice received no intervention. At 1, 7 and 14 days after modeling, peripheral blood cells and bone marrow mononuclear cells were counted, and mitochondrial ultrastructure of the bone marrow was observed by transmission electron microscopy. After Lin- Sca-1+ c-Kit+ (LSK) cells were sorted from bone marrow cells, which were hematopoietic stem cells. The mitochondrial membrane potential, levels of reactive oxygen species, and the expression levels of Bcl 2, Bax, cleaved caspase-9, cleaved caspase-3 and p38 in LSK cells were detected. RESULTS AND CONCLUSION: Compared with the control group, in the model and treatment groups, there were obvious abnormalities in the peripheral blood routine test was the number of bone marrow mononuclear cells was decreased significantly, the mitochondrial number, mitochondrial membrane potential and Bcl 2/Bax ratio were decreased, and the level of reactive oxygen species and the expression of cleaved caspase-9, cleaved caspase-3 and p38 in LSK cells were significantly increased (P < 0.05). After angelica polysaccharide treatment, all above results were significantly improved as compared with the model group (P < 0.05). To conclude, angelica polysaccharide might improve hematopoietic function by up-regulating the mitochondrial membrane potential, increasing the mitochondrial membrane stability and reversing the levels of apoptotic factors.
ABSTRACT
Objective To investigate effects of angelica polysaccharide on learning and memory abilities, Ach, ChAT, AChE, SOD, MDA in serum, APP and Aβ1-42 in hippocampus in model rats with Alzheimer disease (AD); To explore the mechanism of angelica polysaccharide for the treatment of AD. Methods Seventy SPF Wistar rats were selected for learning and memory ability by water maze. 10 rats were randomly selected (half female and half male) as sham-operation group, and the others were injected with Aβ25-35 by stereotatic techniques, copying AD model rats. 50 rats for learning and memory ability by water maze were successfully divided into model group, positive group, angelica polysaccharide low-, medium-, and high-dose groups, with 10 rats in each group. Rats in model group and sham-operation group were given normal saline for gavage, while rats in medication groups were given relevant medicine for gavage, 2 mL/(100 g?d), for 28 d. The learning and memory ability of rats in each group was tested by Morris water maze during 25-28 days, and the contents of Ach, ChAT, AChE, SOD, MDA in serum and APP and Aβ1-42 in hippocampus were determined. Results Compared with the sham-operation group, the escape latent period of model group was significantly prolonged in place navigation experiment; the target quadrant time was shortened; the latent time for the first time to reach the original escape platform was longer in spatial probe test; the residence time of crossing the original platform position and the target quadrant was shorter; the levels of Ach, the activity of ChAT and SOD in serum decreased; the levels of MDA, the activity of AChE in serum increased; the levels of APP and Aβ1-42 in hippocampus increased, with statistical significance (P<0.05, P<0.01). Compared with model group, the escape latent period of each medication group was shortened in different degrees after the intervention treatment; the residence time of target quadrant was prolonged; the latent time for the first time to reach the original escape platform was shortened; the number of cross platform increased; the levels of Ach, the activity of ChAT and SOD in serum increased; the levels of MDA and the activity of AChE in serum decreased; the levels of APP and Aβ1-42 in hippocampus significantly decreased, with statistical significance (P<0.05, P<0.01). Conclusion Angelica polysaccharide may effectively improve the learning and memory of ability of AD model rats to improve anti-free radical oxidation and promote Aβ metabolism and promote learning and memory ability of AD model rats, which have some preventive and therapeutic effects on AD.
ABSTRACT
Objective:To investigate the protective effect of Angelica polysaccharide on the glaucoma-induced retinal neuronal injury,and to elucidate its mechanism.Methods:Eighty-four SD rats were randomly divided into control group,model group,positive drug group,low,middle and high doses of Angelica polysaccharide groups.The model of rat glaucoma was established by sclera sclerotherapy.The intraocular pressures (IOP) of rats in each group were measured.The superoxide dismutase (SOD) activities,malondialdehyde (MDA) contents and nitric oxide (NO) levels in retina tissue of the rats were measured by colorimetric assay.The expression levels of Caspase-3 mRNA and protein in retina tissue of the rats were observed by RT-PCR and Western blotting method.Results:Compared with control group,the IOP of the rats in model group at each time point were significantly increased (P＜0.05);compared with model group,the IOP of the rats in different doses of Angelia polysaccharide groups were significantly decreased (P＜0.05);compared with low dose of Angelica polysaccharide group,theIOP of the rats in high dose of Angelica polysaccharide group was significant decteased (P＜0.05).The levels of MDA and NO in the retina tissue of the rats in model group were higher than those in control group (P＜0.05),but the SOD activities were decreased (P＜0.05);compared with model group,the levels of MDA and NO in retinal tissue of the rats in different doses of Angelica polysaccharide group were significantly decreased (P＜0.05),and the SOD activities were significantly increased (P＜0.05);compared with low dose of Angelica polysaccharide group,the above indexed had significant differences in high dose of Angelica polysaccharide group (P＜0.05).Compared with model group,the expression levels of Caspase-3 mRNA and protein in different doses of Angelica polysaccharide groups were significantly decreased (P＜0.05);compared with low dose of Angelica polysaccharide group,the expression levels of Caspase-3 mRNA and protein were significantly decreased (P ＜ 0.05).Conclusion:Angelica polysaccharide has protective effect on the retinal tissue of glaucoma model rats,which can decrease the levels of MDA and NO in retina tissue and increase the activity of SOD and decrease the expression levels of Caspase-3 mRNA and protein in retina tissue.It may be one of the mechanisms of polysaccharide protecting in retinal tissue nerve cells.
ABSTRACT
Objective: To investigate the protective effect of Angelica polysaccharide on the glaucoma-induced retinal neuronal injury, and to elucidate its mechanism. Methods: Eighty-four SD rats were randomly divided into control group, model group, positive drug group, low, middle and high doses of Angelica polysaccharide groups. The model of rat glaucoma was established by sclera sclerotherapy. The intraocular pressures (IOP) of rats in each group were measured. The superoxide dismutase (SOD) activities, malondialdehyde (MDA) contents and nitric oxide (NO) levels in retina tissue of the rats were measured by colorimetric assay. The expression levels of Caspase-3 mRNA and protein in retina tissue of the rats were observed by RT-PCR and Western blotting method. Results: Compared with control group, the IOP of the rats in model group at each time point were significantly increased (P<0. 05); compared with model group, the IOP of the rats in different doses of Angelia polysaccharide groups were significantly decreased (P<0. 05); compared with low dose of Angelica polysaccharide group, the IOP of the rats in high dose of Angelica polysaccharide group was significant decteased (P<0. 05). The levels of MDA and NO in the retina tissue of the rats in model group were higher than those in control group (P<0. 05), but the SOD activities were decreased (P<0. 05); compared with model group, the levels of MDA and NO in retinal tissue of the rats in different doses of Angelica polysaccharide group were significantly decreased (P<0. 05), and the SOD activities were significantly increased (P<0. 05); compared with low dose of Angelica polysaccharide group, the above indexed had significant differences in high dose of Angelica polysaccharide group (P<0. 05). Compared with model group, the expression levels of Caspase-3 mRNA and protein in different doses of Angelica polysaccharide groups were significantly decreased (P<0. 05); compared with low dose of Angelica polysaccharide group, the expression levels of Caspase-3 mRNA and protein were significantly decreased (P < 0. 05). Conclusion: Angelica polysaccharide has protective effect on the retinal tissue of glaucoma model rats, which can decrease the levels of MDA and NO in retina tissue and increase the activity of SOD and decrease the expression levels of Caspase-3 mRNA and protein in retina tissue. It may be one of the mechanisms of polysaccharide protecting in retinal tissue nerve cells.
ABSTRACT
Objective: To investigate the protection of angelica polysaccharide (APS) on hippocampal neuron in rats with cerebral ischemia-reperfusion (I/R) injury. Methods: The model of cerebral I/R injury was established by suture method in rats. A total of 50 rats were randomly divided into five groups: Sham-operation, cerebral I/R injury, high-dose APS (200 mg/kg), mid-dose APS (100 mg/kg), and low-dose APS (50 mg/kg) groups. APS was ig administrated 3 d before operation. At 24 h after reperfusion, learning and memory function was detected by step down test, the apoptosis of hippocampal neuron was observed by terminal deoxylnucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the expression of cleaved caspase-3, bcl-2, and bax in the hippocampus of rats was measured by enzyme-linked immunosorbent assay (ELISA). Results: Compared with those in the Sham-operation group, the learning and memory function was notably impaired in the I/R injury group, the number of errors increased. The apoptosis of hippocampal neuron increased and the expression of cleaved caspase-3, bcl-2, and bax in the hippocampus remarkably increased in the I/R injury group. The APS could significantly improve the learning and memory function of rats with the cerebral I/R injury and remarkably delay the decrease of the number of errors and the decrease of the apoptosis rate in the hippocampus of rats with the cerebral I/R injury. And the APS could also cause a significant down-regulation of cleaved caspase-3 and bax expression, while up-regulation of bcl-2 expression in hippocampus of rats with the cerebral I/R injury. Conclusion: APS has a neuroprotection on rats with the cerebral I/R injury. The neuroprotective mechanism of APS may involve in the inhibition of the neuronal apoptosis.
ABSTRACT
Objective To explore the relative signal transduction pathway of angelica polysaccharide(APS) inducing K562 cells toward erythroid differentiation. Methods K562 cells were divided into control group and ASP group, the control group cells were routinely cultured and APS group cells were treated with APS(final concentration is 100-500mg/L). By using of benzidine staining and spectrophotometry, the characteristics of erythroid differentiation of K562 cell induced by APS were detected;By using of laser confocal microscopy, the distribution of JAK_2 and STAT_5 in K562 cells was observed;By using of Western blotting, the expression of JAK_2 and STAT_5 in nucleus and cytoplasm of K562 cells was detected, By using of imm～uno～pre～cipi～ta～tion, the phosphorylation change of JAK_2 in cytoplasm was tested. Results After being induced by APS, the benzidine staining positive rate of K562 was increased.With increasing the concentration of APS, hemoglobin synthesis in K562 cells was promoted accordingly. After being cultured for 12 hours, 24 hours and 48 hour, the expression of STAT_5 in nucleus of K562 cells induced by APS was significantly higher than that of control group, however, expression of STAT_5 in cytoplasm of K562 cell induced by APS was obviously lower. The expression of JAK_2 in K562 cells was not different between APS group and control group, but the JAK_2 phosphorylation level of APS group was much higher than that of control group.Conclusion APS can induce erythroid differentiation of K562 cells, and the mechanisms may be that APS can promote the phosphorylationthe of JAK_2, then stimulating nuclear translocation of STAT_5.
ABSTRACT
Objective To investigate the effect of angelica polysaccharide (APS) on bioactivity of bone marrow macrophage (BMM?) and its relationship to hematopoietic regulation, for clarifying the hematonic mechanism of Angelica sinensis. Methods The techniques of hematopoietic progenitor cell culture and BMM? culture in vitro, biological assay of hematopoietic growth factor (HGF) in culture media of BMM?, immunocytochemistry, and nucleic acid in situ hybridization were used. Results The culture supernatant of BMM? induced by APS can enhance the CFU-Mix, CFU-E, CFU-GM; the expression of erythropoietin (EPO), GM-CSF, IL-3, IL-6 protein in BMM? induced by APS was much stronger than that in the control group at different levels; the expression of EPO mRNA and GM-CSF mRNA in BMM? induced by APS were intensified. Conclusion APS may directly and/or indirectly stimulate the BMM? in hematopoietic inductive microenvironment to accelerate the synthesis and secretion of hematopoietic regulation factors on the basis of gene and protein level, such as EPO, GM-CSF, IL-3, IL-6, which in turn to promote the proliferation and differentiation of CFU-Mix, CFU-E, and CFU-GM.
ABSTRACT
Object To investigate the effect of Angelica polysaccharide (APS) on the induction of chronic myelocytic leukemia cells into chronic myelocytic leukaemia dendritic cells (CML-DCs). Methods Bone marrow monocytes from CML patients were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4 combined with APS in each concentration (50, 100, 200 mg/L), respectively. The morphotype of CML-DCs was identified by optical microscope or electron microscope, CML-DCs viability was calculated by Trypan Blue exclusion. The phenotype of CML-DCs (CD 80, CD 86, and CD 83) was identified by flow cytometry. The capability of stimulating auto-lymphocyte or allo-lymphocyte proliferation was tested with mixed leukocyte reaction (MLR). Results Bone marrow monocytes from CML patients, which were cultured in GM-CSF/IL-4 or in GM-CSF/IL-4/APS showed typical morphotype and expressed the high level phenotype of CML-DCs. The capability of proliferation and the survival rate of CML-DCs were enhanced markedly and the expression of CD 83, CD 80, and CD 86 on CML-DCs were significantly increased when CML-DCs were cultured in GM-CSF/IL-4/APS. The capability of stimulating lymphocyte proliferation was more competent in 100 mg/L APS group. Conclusion The expression of CD 83, CD 80, and CD 86 on CML-DCs cultured in GM-CSF/IL-4/APS is significantly higher than those in GM-CSF/IL-4. The capability of CML-DCs of stimulating lymphocyte proliferation is more potential in GM-CSF/IL-4/APS than in GM-CSF/IL-4. APS can promote the induction and mature of CML-DCs cultured in IL-4 and GM-CSF.
ABSTRACT
Object To investigate the effects of Angelica polysaccharide (AP 0) and its sulfates (APS) on blood coagulation and platelet aggregation. Methods Infrared turbidimetric method was used to estimate platelet aggregation, prothrombin time (PT) and thrombin time (TT), actiated partial thromboplastin time (APTT); bleeding time (BT) was measured by cutting the mouse's tail and coagulate time (CT) was measured by glass method sheet; hemorheology was measured with heparin as anticoagulant.Results The AP 0 and APS prominently enhanced the platelet aggregation at 5 min (PAG ), while showed less effects on the maximum platelet aggregation (PAG ); they, especially APS 1, also markedly prolonged CT but shortened BT, both AP 0 and APS 1 significantly prolonged TT and APTT in all experimental dosage, while only APS 1 in high dosage (8.46 mg/kg) significantly prolonged PT; AP 0 increased the aggregation index (AI) of RBC and the whole blood viscosity (? b) at shear rate of 5 s -1 at a dose of 2.00 mg/kg, however, unlike AP 0, APS 1 decreased AI and ? b at shear rate of 5 s -1 and 230 s -1 . Conclusion These results suggest that AP 0 and APSs have potent anticoagulant and haemostasis effects. The haemostasis effect is related to promoted platelet aggregation. Ameliorative effect of Angelica polysaccharide sulfate on hemorheology is better than that of Angelica polysaccharide itself.
ABSTRACT
Objective:To study modern biological mechanism for hematonic and antitumor of Angelica Sinensis (oliv) Diels.Methods:By using the techniques of hematopoietic cells culture in vitro,morphological observation,flow cytometry and immunocytochemistry,the effect of Angelica polysaccharide (APS) on proliferation and differentiation of human CFU-GM and promyelocytic leukemia cell line (HL-60) was studied.Results:APS could markedly promote the colony formation of CFU-GM and could significantly inhibit the proliferation of HL-60 in vitro.The inhibitory effect is concerned with the concentration of APS.APS may prevent HL-60 from the Go phase entering the S/G2+M phase,thus prohibit the synthesis of DNA.APS may also promote HL-60 entering the process of apoptosis.Conclusion:APS may not only promote normal hematopoiesis,but also inhibit the proliferation of leukemia cell and it can be a natural inducer for therapy of malignant tumor.