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OBJECTIVE To investigate the effects of Angelica sinensis polysaccharide on the apoptosis of cardiomyocytes in diabetic KK-Ay mice. METHODS KK-Ay mice were randomly divided into model group， metformin group （200 mg/kg） and A. sinensis polysaccharide high-dose， medium-dose and low-dose groups （400， 200 and 100 mg/kg）； C57BL/6J mice were included in blank group， with 8 mice in each group. Each group was given relevant medicine intragastrically or normal saline， once a day， for consecutive 4 weeks. After the final administration， the levels of fasting glucose， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C） and insulin （INS） were detected； the protein expressions of B-cell lymphoma 2 （Bcl-2）， cleaved- caspase-3， apoptosis signal-regulated kinase 1 （ASK1）， phosphorylated c-Jun N-terminal kinase （p-JNK）， phosphorylated inositol- requiring enzyme 1α （p-IRE1α） in myocardium， and apoptosis in cardiomyocytes were also detected. RESULTS Compared with model group， the fasting glucose， TC and LDL-C content， apoptotic rate of cardiomyocyte， protein expressions of p-JNK and p- IRE1α， ASK1， cleaved-caspase-3 were significantly lower in the metformin group and A. sinensis polysaccharide medium-dose， high-dose groups； INS level and relative expression of Bcl-2 protein were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS A. sinensis polysaccharide can improve the levels of blood glucose and blood lipid and inhibit cardiomyocyte apoptosis in diabetic KK-Ay mice， and the mechanism may be related to the inhibition of IRE1/ASK1/JNK signaling pathway.
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Objective To investigate the effect of Angelica Sinensis polysaccharide (ASP) on proliferation, differentiation and transplantation of human leukemia stem cells (LSCs) . Methods 1. Effect of angelica sinensis polysaccharides on proliferation of CD34
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OBJECTIVE@#This study aims to evaluate the effect of Angelica sinensis polysaccharide (ASP) on the osteogenic differentiation of the bone marrow mesenchymal stem cells (BMSCs) of rats with high glucose levels.@*METHODS@#Rat BMSCs were isolated and identified by osteogenic and adipogenic differentiation. Then, the BMSCs were divided into three groups as follows: normal control group (5.5 mmol·L⁻¹ glucose), high glucose group (25.5 mmol·L⁻¹ glucose), and ASP+high glucose group (25.5 mmol·L⁻¹ glucose +40 mg·L⁻¹ ASP). The proliferation activities of the BMSCs were detected by CCK8. Alizarin red staining, and alkaline phosphatase activity were used in the examination of osteogenic activity. Quantitative real time-polymerase chain reaction was used to detect the expression levels of the osteogenic genes (Runx2, Osx, OCN, Col-Ⅰ) and the key factors of Wnt/β-catenin signal pathway (CyclinD1, β-catenin). In vivo, a type 2 diabetes rat model was established. The rats were divided into three groups, namely, the normal control group (normal rats), diabetes group (diabetic rats), diabetes+ASP group (diabetic rats, ASP feeding). Then, the tibia bone defect was established. The repair of bone defects in each group was observed through histological examination.@*RESULTS@#The proliferation of BMSCs was higher in the high glucose group and ASP+high glucose group than in the normal control group (P0.05). The number of calcium nodules of BMSCs; alkaline phosphatase activity; and the mRNA expression of Runx2, OCN, Osx, Col-Ⅰ, CyclinD1, β-catenin in the high glucose group were lower than those in the normal control and ASP+high glucose groups (P0.05). The bone mass was significantly lower in the bone defect of the diabetes group than in the bone defect of the normal control or diabetes+ASP group (P0.05).@*CONCLUSIONS@#ASP can promote the osteogenic differentiation of rat BMSCs under high glucose culture and induce bone regeneration in rats with type 2 diabetes. These features may be related to the activation of the Wnt/β-catenin signaling pathway.
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Animals , Rats , Angelica sinensis , Chemistry , Bone Marrow Cells , Cell Differentiation , Cells, Cultured , Diabetes Mellitus, Experimental , Diabetes Mellitus, Type 2 , Glucose , Mesenchymal Stem Cells , Osteogenesis , Plant Extracts , Pharmacology , Polysaccharides , PharmacologyABSTRACT
Objective To investigate the protective effects of Angelica sinensis and Angelica sinensis polysaccharide against liver and kidney injury induced by irradiation of SD rats.Methods Thirty-two SD rats were randomly divided into four groups:control group,model group,Angelica decoction group and Angelica sinensis polysaccharide group.After 14 days of routine feeding,the rats were treated by intragastric administration once a day for 7 days.From the 8th day,the rats received whole body X-ray irradiation for 2 days.The total absorbed dose was 6Gy,and the dose rate was 92.26cGy/min.The rats were killed and the blood were collected from the femoral artery 3 days after irradiation.The content of superoxide dismutase (SOD),malondialdehyde (MDA),glutathione peroxidase (GSH-Px) and lipid peroxide (LPO) in the liver and kidney tissues was detected by colorimetric analysis.The pathological changes of the liver and kidney were observed by HE staining under microscope.The expression of Nrf2 protein in the liver and kidney of the rats was detected by Western blotting.Results Compared with control group,the weight of the liver and kidney significantly increased,the activities of SOD and GSH-Px significantly decreased,the contents of MDA and LPO significantly increased,and the liver and kidney cells showed obvious edema,and the content of Nrf2 in the liver and kidney increased in the model group.Compared with model group,the weight of the liver and kidney decreased,SOD and GSH-Px activities significantly increased,the contents of MDA and LPO significantly decreased,edema of the liver and kidney significantly reduced,the expression of Nrf2 in the liver and kidney tissues decreased in Angelica decoction group and Angelica sinensis polysaccharides group,while there was no difference between Angelica sinensis polysaccharide group and Angelica decoction group.Conclusion Angelica sinensis and Angelica polysaccharides have a good protective effect against liver and kidney injury induced by radiation in SD rats,which is implemented mainly through influencing the expression of free radicals.The protective effect of Angelica against radiation injury is achieved mainly by Angelica polysaccharide.
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Aim To investigate the protective effects of Angelica sinensis polysaccharides on bone marrow and spleen injury in SD rats induced by X-ray radiation.Methods Forty male SD rats were randomly divided into blank group,model group,low dose group of Angelica polysaccharide (63.5 mg · kg-1),Angelica polysaccharide middle dose group (127 mg · kg-1),and Angelica polysaccharide high dose group (254 mg · kg-1).The medicine group was treated daily with different concentrations of Angelica polysaccharide 2 mL gavage once,blank group and model group with an equal amount of distilled water instead.After a continuous administration of 7 d,X-ray irradiation was performed on rats except the blank group,with continuous exposure of 2 d,and the total radiation was at a dose of 6 Gy,then 3 d post the last radiation,the rats were sacrificed after anesthesia by bloodletting.The general quality of life,bone marrow nucleated cells,blood routine,spleen index,serum interferon gamma (IFN-γ)and interleukin-4 (IL-4) contents in each group of rats were detected.Pathological changes of spleen were observed under microscope.Results Compared with the blank group,the quality of life,bone marrow nucleated cells of rats in the model group,the number of white blood cells,spleen index,INF-γ and IL-4 expression significantly decreased,but the number of erythrocytes and hemoglobin increased significantly,and the difference was statistically significant (P ＜ 0.05).HE staining showed the decrease in splenic corpuscle atrophy,germinal center,white pulp and red pulp structural disorder,and the number of lymphocytes.Compared with the model group,Angelica polysaccharide group's survival quality,bone marrow nucleated cell number,the number of white blood cell,spleen index,INF-γand IL-4 expression markedly increased,but the number of erythrocytes and hemoglobin decreased significantly,and the difference was statistically significant (P ＜ 0.05).HE staining showed that pathological injury of spleen was alleviated,spleen body was visible,germinal center,red pulp and white pulp boundaries were still visible,and the number of lymphocytes increased.Conclusions Angelica sinensis polysaccharide can protect the bone marrow and spleen of SD rats induced by radiation,and the mechanism may be achieved by reducing the damage to hematopoietic cells and immune cells in the bone marrow and spleen,and replenishing blood and body fluid body.
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Objective:To optimize the ultrasonic extraction process of Danggui Baogan capsules. Methods:The ultrasonic extrac-tion process of Danggui Baogan capsules was optimized by L9 (34 ) orthogonal test with the contents of ferulic acid and angelica sinensis polysaccharide as the indices and the amount of 30% ethanol, ultrasonic duration and extraction times as the influencing factors. Re-sults:The best ultrasonic extraction conditions were as follows: adding 12-fold amount of 30% ethanol and extracted four times with 1. 5 hours for each time. Conclusion:The ultrasonic extraction process is simple and reproducible, and suitable for the industrial pro-duction.
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Objective: To explore the hereditary basis of secondary metabolic biosynthesis in Angelica Sinensis Radix (the roots of Angelica Sinensis) and identify the gene sequence of active substance in Angelica Sinensis Radix. Methods: The samples of Angelica Sinensis Radix were collected in Min County of Gansu Province and were identified by relevant experts. Total RNA was extracted from the heads and tails of Angelica sinensis. Sequencing library was constructed and qualified. Then, Double-ended sequencing was accomplished in Illumina HiSeq 2000 device. Transcriptome characteristic of Angelica Sinensis Radix was analyzed by relevant bioinformatics. Results: We obtained 66 431 540 primitive sequences of Angelica Sinensis Radix by Illumina HiSeq 2000 high-flux sequencing and annotated 30 432 unigenes by bioinformatics. The important economic crop distributed in Angelica Sinensis Radix sequences are Vitis vinifera, Ricinus communis, Populus trichocarpa, Glycine max, Medicago truncatula, Arabidopsis thaliana, and Nicotiana tabacum through comparing with 939 species sequences in Uniprot protein library. It was suggested that 127, 69, 70, and 94 unigenes expressed in Angelica Sinensis Radix were respectively mapped to the pathway of phenylpropanoid biosynthesis, N-glycan biosynthesis, flavonoid biosynthesis, and folate biosynthesis. These unigenes are related to the biosynthesis of important active substances, for example ferulic acid, polysaccharide, flavonoid, and folic acid in Angelica Sinensis Radix. Conclusion: Unigenes explored in the present paper are involved in the biosynthesis of major pharmaceutical substances, suggesting the genetic basis of Angelica Sinensis Radix.
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Objective: To explore the effects of Angelica sinensis polysaccharide (ASP) on Wnt/β-catenin signaling pathway in hematopoietic stem cell (HSC) of aging model mice Methods: Male Thirty C57BL/6J mice aging 6-8 weeks were randomly divided into normal group, aging model group, and ASP aging model group (ten in each group). After 2 d of finishing the treatment, magnetic activated cell sorting (MACS) were applied to the HSC from mouse bone marrow respectively. The ratio of the SA-β-Gal staining positive HSCs were counted; The capability of colony formation was examined by CFU-Mix cultivation; The distribution of ROS levels was analyzed by flow cytometry (FCM) and laser scanning confocal microscope assess; The content of advanced glycosylation end products (AGEs) was detected by Elisa; The proteins of β-catenin in cytoplasm, GSK-3β in nucleus, Phospho-GSK-3β, and TCF-4 were detected by Western blotting. Results: Compared with the normal group, the percentage of SA-β-Gal, the product of ROS positive cells and AGEs in the aging model group were significantly increased; The expression of β-catenin in cytoplasm, β-catenin in nucleus, Phospho-GSK-3β, and TCF-4 were evidently up-regulated; The colony formation of CFU-Mix was markedly decreased; The expression of GSK-3β was evidently down-regulated. Compared with the aging model group, in ASP aging model group, the percentage of SA-β-Gal, the product of ROS positive cells, and AGEs were significantly decreased; The expression of β-catenin in cytoplasm and nucleus, Phospho-GSK-3β, and TCF-4 were evidently down-regulated; The colony formation of CFU-Mix was markedly increased; The expression of GSK-3β was evidently up-regulated. Conclusion: ASP can protect HSC from aging by antagonizing D-galactose, and the mechanism may be ASP inhibiting the excessive activation of Wnt/β-catenin signaling pathway.
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Objective: To investigate the mechanisms on the regulation of telomere and telomerase in the process of senescence induction of human-derived CD34+CD38- leukemia stem cells (LSC) subpopulation by Angelica sinensis polysaccharide (ASP). Methods: The human-derived CD34+CD38- LSC subpopulation was isolated from acute myelogenous leukemia bone marrow mononuclear cells by magnetic activated cell sorting. The inhibition of ASP on CD34+CD38- LSC subpopulation proliferation was detected by CCK-8 assay. The percentage of senescent cells was detected by SA-β-Gal staining. The colony-formed ability was detected by Colony-forming Assay. The levels of telomerase activities and telomerase reverse transcriptase (TERT) gene expression were performed by TRAP-PCR and quantitative RT-PCR, respectively. The changes of telomere length were tested by Southern blotting assay. Results: The CD34+CD38- LSC subpopulation could be effectively isolated by MACS. The purity of CD34+CD38- LSC population is up to (91.15 ± 2.41)%; The cells showed the features of well-stacked morphology, high transparency, well refraction under inverted phase contrast microscope. ASP had a remarkable dose-dependent inhibition on CD34+CD38- LSC proliferation in vitro culture (P < 0.05). The number of SA-β-Gal staining positive cells had been increased compared to the cells in control group (P < 0.01), a decrease in colony-forming abilities (P < 0.01), a decrease level on TERT gene and telomerase activities (P < 0.05), and a shorter length on telomere of CD34+CD38- LSC after 40 μg/mL ASP co-culture for 48 h (P < 0.05). Conclusion: ASP could induce the senescence of human derived leukemia bone marrow CD34+CD38- LSC via regulating the cell telomere system in vitro co-culture.
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Objective To isolate and purify polysaccharide from Angelica sinensis by ultrafiltration technology. Methods Extract of polysaccharide of Angelica sinensis was ultrafiltrated with different aperture membrane, the separation parameters on membrane separation efficiency of polysaccharide of Angelica sinensis were optimized by and orthogonal test. Results Polysaccharide of Angelica sinensis was separated with 200, 100, 50, 20 kD membrane. The contents of polysaccharide was 30.47% before ultrafiltration. The main component of Angelica sinensis polysaccharide was the fraction with 200 kD (65.42%). The optimum condition of ultrafiltration was obtained as follow:one times of extract volume, 35 ℃, 0.3 MPa. Conclusion The technology was simple and available, and the content of Angelica sinensis polysaccharide was higher.
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AIM:To explore the therapeutic effect of dexamethasone Angelica sinensis polysaccharide prodrug(DEX-AP) on trinitrobenzene sulfonic acid(TNBS) induced ulcerative colitis(UC) in rats and its side effects.METHODS: The experimental UC rats were induced by clusis of the solution of TNBS in 45% alcoho1(50(mg?ml~(-1))).The UC rats were orally administrated with(0.25)(?mol?kg~(-1)?d~(-1)) DEX and(0.05),(0.25),(1.25)(?mol?kg~(-1)?d~(-1)) DEX-AP(calculated by carried DEX in DEX-AP) for 7 days,respectively.The rats were killed after the amount of peripheral blood lymphocyte was counted,then the spleen,thymus and colon were separated and weighted.After the ulcerative area of colon was calculated,the colonic myeloperoxidase(MPO) activity was determined and parts of colon were paraffin sectioned and examined under light microscope by HE stain.RESULTS: After the UC rats were administrated with different doses of DEX-AP for 7 days,the ulcerative area,the weight and the MPO activity of colon reduced significantly.The reduction of MPO activity was correlated to the dose of DEX-AP and the MPO activity with DEX-AP at the doses of(0.25),(1.25)(?mol?kg~(-1)?d~(-1)) reduced more significantly than that with DEX at the the dose of(0.25)(?mol?kg~(-1)?d~(-1)).The number of peripheral blood lymphocyte,spleen weight and thymus weight of UC rats reduced significantly at the dose of(0.25)(?mol?kg~(-1)?d~(-1)) DEX(P
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AIM: To investigate the immunoloregulation effect of Angelica polysaccharide extracted from Angelica Sinensis (Oliv.) Diels. METHODS: Index of immune organs were weighed and calculated. Phagocytosis of mononuclear macrophage (M) were determined with carbon particle clearance test. Spectrophotography was used to estimate levels of serum hemolysis IgG, IgM. The cell proliferation was measured by MTT assay. RESULTS: In certain doses (10-100 mg/kg), Angelica polysaccharide significantly increased phagocytic function and spleen index in immunosuppression mice caused by hydrocortisone, while showed no obviouse influnence on thymus index. It also remarkble decreased levels of serum hemolysis IgG, IgM. In vitro experiments, Angelica polysaccharide could increase the proliferation of mouse spleen cell and macrophage. CONCLUSION: These results suggest that Angelica polysaccharide has potent enhancement on non specific immunity, however, it could suppress humoral immunity.