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OBJECTIVE To study the tocolysis effects of Angelica sinensis polysaccharides on threatened abortion model rats and their impacts on Th1/Th2 balance by regulating the phosphoinositide 3-kinase （PI3K）/protein kinase B （AKT） signaling pathway. METHODS Pregnant rats were randomly grouped into the control group， model group， A. sinensis polysaccharide group （200 mg/kg）， PI3K/AKT signaling pathway inhibitor LY294002 group （5 mg/kg）， and A. sinensis polysaccharide+LY294002 group （200 mg/kg A. sinensis polysaccharide+5 mg/kg LY294002）， with 10 rats in each group. Except for the control group， rats in all other groups were given mifepristone （8.3 mg/kg） and misoprostol （100 μg/kg） intragastrically to establish a threatened abortion model， and intragastric or intraperitoneal injection of corresponding drugs. The serum levels of estrogen， progesterone， interferon-γ （IFN-γ）， tumor necrosis factor-α （TNF-α）， and interleukin-4 （IL-4） in each group of rats were detected， and the uterine ovarian index and embryonic mortality rate of rats in each group were measured； the morphology of uterine tissue in rats was observed in each group； Th1/Th2 balance in peripheral blood of rats as well as the expression of PI3K/AKT signaling pathway-related proteins in the uterine tissues of rats in each group were detected. RESULTS Compared with the control group， the uterine tissue of rats in the model group showed pathological damage； the serum levels of estrogen， progesterone and IL-4， uterine ovarian index， peripheral blood Th2 cell ratio， and the ratios of phosphorylated PI3K （p-PI3K）/PI3K and phosphorylated AKT （p-AKT）/AKT in uterine tissue were all decreased （P＜0.05）； the embryo mortality rate， Th1 cell ratio， Th1/Th2 ratio， and serum levels of IFN-γ and TNF-α were increased （P＜0.05）. Compared with the model group， the pathological damage of uterine tissue in the A. sinensis polysaccharide group was reduced， and the above indexes were all improved significantly （P＜0.05）； LY294002 could weaken the effect of A. sinensis polysaccharide on model rats （P＜0.05）. CONCLUSIONS A. sinensis polysaccharides can improve Th1/Th2 imbalance in threatened abortion model rats by activating the PI3K/AKT signaling pathway， thereby inhibiting immune inflammation， and promoting embryo survival.
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Aim To investigate the effects of angelica sinensis polysaccharide (ASP) antagonizing 5-fluorou-raeil (5-FU) on spleen stress erythropoiesis in mice and its related mechanism. Methods C57BL/6J mice aged 6-8 weeks were randomly divided into control group, ASP group, 5-FU group and ASP + 5-FU group. The mouse body weight during the modeling pe-riod was recorded, and peripheral blood routine and the number of mononuclear cells in the bone marrow of femur were measured. Histopathology of spleen was de-tected, also the index and cellularity of spleen were analyzed. BFU-E of spleen mononuclear cells was counted. The number of F4/80
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Objective To study the protective role of Angelica sinensis polysaccharides (ASP) on liver injury of SD rat caused by X-ray radiation. Methods Forty SD rats after 14 d of conventional breeding were randomly divided into control group, model group, ASP high-, medium-, and low-dose (254, 127, and 63.5 mg/kg) groups. The intervention groups were given different concentration of ASP lavage, and the control and model groups were given the same amount of distilled water instead, once daily, 2 mL/time, for continuous 7 d. On day 8, except the control group, the rest rats were given X-ray irradiation of total body after 30 min of lavage, for 2 d continuous exposure, and total radiation dose was 6 Gy. After 72 h of irradiation, femoral artery blood was collected and rats were killed. The activities of serum AST, ALT and the contents of SOD, MDA, GSH-Px, and LPO in liver were detected. HE staining was used to observe morphological changes of liver tissue, Western blotting was used to detect the Nrf2 protein expression in rat liver tissue. Results Compared with the blank group, in the model group the activities of AST and ALT in serum of rats, MDA and LPO content increased significantly, SOD and GSH-Px content significantly decreased, and Nrf2 protein expression increased, with statistically significant difference (P < 0.05). HE staining showed that hepatic nuclei reduces and partly disappears, and liver cells were degenerated. Compared with model group, serum AST and ALT activities of rats in high-, medium-, and low-dose groups decreased, the contents of MDA and LPO decreased, the contents of SOD and GSH-Px in liver tissue increased, and the Nrf2 protein expression decreased, with statistically significant differences (P < 0.05). HE staining showed hepatic shrinking nucleus and cytoplasm vacuoles degeneration. Conclusion The ASP have protective effect on the liver injury of SD rats caused by X-ray radiation, its mechanism is probably though reducing the formation of free radicals in the liver tissue and improving the liver’s ability to remove free radicals.
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Objective:To establish an HPLC-FD method for determining the content of Angelica sinensis polysaccharide (ASP1) to lay foundation for its pharmacokinetic study in rats. Methods: Purified ASP1 was labeled with FITC by the method of Belder and Granath to obtain ASP1-FITC. The tissue samples were treated with 30% trichloroacetic acid and 11% NaOH before injection. The samples were determined by HPLC-FD. A PL aquagel-OH MIXED column was used,and the mobile phase was phosphate buffer( dis-solve NaH2PO32. 34g , Na2HPO34. 33 g and NaCl 11. 70 g into 1000 ml water) with pH of 7. 0. The flow rate was 0. 5 ml·min-1. The excitation and emission wavelengths was set at 495 nm and 520 nm, respectively. Results:The linear calibration curve was within the concentration range of 0. 25-40. 00 μg·ml-1(r=0. 9996)with the lower limit of quantification of 0. 20μg·ml-1 in tissue sam-ples. The extraction recovery of ASP1 was determined at low, medium and high concentration with the recovery of 91. 98%-114. 20%. The intra and inter-day RSDs were lower than 8. 31% and 2. 94%, respectively. Conclusion:The method to determine the content of ASP1 in rats by HPLC-FD with pre-column derivatization has been esablished. It is simple, accurate and reliable, which can be suc-cessfully applied in the study of pharmacokinetics and tissue distribution of ASP1 in rats.
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Objective: To explore the effects and the possible mechanism of combined injection of Angelica sinensis polysaccharide (ASP) and cytarabine (Ara-C) on the bone marrow mononuclear cells (BMMCs) of the transplanted human leukemia mouse model. Methods: K562 cells (2×107)were transplanted into the tail vein of mice to establish the transplanted human leukemia NOD/SCID mouse model. Then the leukemia mice were randomly divided into model, ASP, Ara-C, and ASP + Ara-C groups. After the 30 d transplantation, the mice were ip injected with ASP (200 mg/kg/d), Ara-C (2.5 mg/kg/d), and ASP (200 mg/kg/d) + Ara-C (2.5 mg/kg/d), respectively for 14 d, and the mice in the model group were injected with saline (equal volume and time). The next day after the treatment, the eyeball blood was collected to detect the amount and classification of white blood cells (WBC). The femurs were taken to count BMMCs of each femur. The proliferation of BMMCs was detected by CCK-8; The distribution of cell cycles was analyzed by flow cytometry (FCM); The capability of colony forming was examined by CFU-Mix cultivation; The ratio of the SA-β-Gal staining positive BMMCs was counted; The aging related proteins of P16, Rb, CDK4, and CyclinD1 were detected by Western blotting. Results: Compared with the model group, ASP or Ara-C injected alone and their combined injection can obviously reduce the amount of the peripheral blood WBC, the percentage of neutrophiles, and the number of femur BMMCs; effectively inhibit the proliferation of BMMCs, CFU-Mix forming, and the ratio of S stages; markedly raise the percentage of lymphocytes, ratio of G1 stages, and the percentage of SA-β-Gal positive cells; down-regulate the expression of the aging related proteins of CDK4 and CyclinD1; and up-regulate the expression of P16 and Rb protein. The effects of ASP + Ara-C group were much better than those in the other groups. Conclusion: The aging mechanism of BMMCs for the transplanted human leukemia mice induced by ASP and Ara-C may be ascribed to the regulation of the expression of the aging related proteins of P16, Rb, CDK4, and CyclinD1. Our research provides a new idea to treat leukemia in clinic.
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Objective To investigate to the role of Angelica sinensis polysaccharides (ASP) against hepatic encephalopathy and explore the probable mechanism. Methods Sprague-Dawley rats were established into carbon tetrachloride (CCl4)-induced liver cirrhosis,then treated with distilled water (negative control),lactulose (positive control),ASP solution (10,20 and 40 g/L respectively,intragastrically at dose of 10 ml.kg-1.d-1) for 14 days. On 15th day,all the survival rats were intraperitoneally injected with endotoxin (3 mg/kg) once,then in the following 12 h,the rats that fall into coma were counted and the time period of coma was recorded. The blood plasma concentrations of ammonia,serum glutamate-pyruvate transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) were determined. The apoptotic cerebral cells were dyed in situ by TUNEL. Glial fibrillary acidic protein (GFAP) in astrocytes was determined by immunohistochemical method. Then we counted the apoptotic cerebral cells and GFAP positive cells under light microscope. Results The survival rats in distilled water group were highest in the concentrations of ammonia,GPT and GOT,the amount of apoptotic cerebral cells,and coma rate than the rats in other groups. The rats in 40 g/L ASP group had the higher ammonia level,GOT concentration,coma rate than those in lactulose group. The rats in 3 ASP groups all had less apoptotic cerebral cells than those in lactulose group. As to GFAP,distilled water group had the lowest immunological activity,and the GFAP immunological positive cells of 3 ASP groups were all more than those in lactulose group. Conclusion ASP,even more effective than lactulose,can protect rat brain function and delay the occurrence of hepatic encephalopathy,whose effects may be related to the decrease of blood ammonia,the amelioration of liver function as well as the protection of astrocyte function and neuronic activity.
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AIM: To explore the transport and delivery of active drug from dexamethasone-angelica sinensis polysaccharides prodrug in the gastrointestinal tract of rats. METHODS: Dexamethasone and the prodrug were orally administered to rats at the dose of 1.96 mg?kg~ -1 (calculated by carried dexamethasone). The drugs in the plasma and contents of different parts of the rats' gastrointestinal tract were determined by high performance liquid chromatography (HPLC). RESULTS: Dexamethasone carried by the prodrug was mainly released in the contents and mucosa of cecum and colon after oral administration of the prodrug. The absorption of released dexamethasone was reduced significantly. The peak time, peak concentration and AUC were 7.2 h , 42 ?g?L~ -1 and 334 ?g?h?L~ -1 , respectively. However, free dexamethasone was found mainly in the contents and mucosa of the stomach, proximal and distal small intestine after oral administration. The peak time, peak concentration and AUC were 2.2 h, 2 120 ?g?L~ -1 and 11 875 ?g?h?L~ -1 , respectively. CONCLUSION: Dexamethasone can be specifically delivered to the cecum and colon by using dexamethasone- angelica sinensis polysaccharides prodrug. The absorption of dexamethasone was reduced significantly and the drug concentration in colon was increased significantly. The prodrug has a potential in the treatment of colitis.