Effects of astragalus polysaccharides on improvement of liver and kidney injury and regulation of intestinal flora structure in cadmium-exposed rats / 中国药理学通报

Aim To investigate the effects of astragalus polysaccharides on the improvement of liver and kidney injury and the regulation of intestinal flora structure in cadmium exposed rats. Methods Rats exposed to cadmium were established by intraperitoneal injection of CdCl2. After continuous intragastric administration of astragalus polysaccharides for five weeks, urine, liver, kidney and feces were collected. The cadmium residues in urine, liver and kidney were detected by Graphite Furnace Atomic absorption spectrometry, the pathological changes of liver and kidney were observed by HE staining, and Illumina PE250 sequencing and bioinformatics software were used to analyze the structure of intestinal flora. Results After intraperitoneal injection of CdCl2, the accumulation of cadmium in urine, liver and kidney increased significantly, some liver and kidney cells showed pathological damage such as swelling, necrosis and inflammatory cell infiltration. Chao, ace and shannon indexes decreased significantly, while simpson index increased significantly. The number of OTU decreased. And the abundance of Ruminococcus, Bacteroides, Flavonifractor, Roseburia and Elusmicrobium decreased significantly, but Lactobacillus, that of Lachnospiracea_incertae_sedis, Parasutterella, Clostridium XlVb, Clostridium XI, Integinimonas and Fusobacterium increased significantly. Compared with the normal control group, the differences was statistically significant(P<0.05 or P<0.01). After intragastric administration of astragalus polysaccharides, cadmium accumulation in urine, liver and kidney decreased significantly, liver and kidney cell damage alleviated, and inflammatory cell infiltration reduced. Chao, ace and shannon index increased markedly, and simpson index decreased significantly. OTU number increased. And the bundance of Prevotella, Bacteroides, Parasutterella, Elismicrobium and Barnesiella raised significantly, that of Ruminococcus, Oscillibacter, Flavonifractor, Clostridium XlVa, Roseburia, Lactobacillus, Ruminococcus2, Lachnospiracea_incertae_sedis, Clostridium IV, Clostridium XlVb, Clostridium XI and integinimonas decreased significantly, which was statistically significant compared with the group exposed to cadmium alone(P<0.05 or P<0.01). Conclusions Astragalus polysaccharides may improve liver and kidney injury by reducing cadmium accumulation and regulating the structure of intestinal flora in cadmium exposed rats.

Anti-obesity and Gut Microbiota Modulation Effect of Astragalus Polysaccharides Combined with Berberine on High-Fat Diet-Fed Obese Mice / 中国结合医学杂志

OBJECTIVE@#To investigate whether astragalus polysaccharides (APS) combined with berberine (BBR) can reduce high-fat diet (HFD)-induced obesity in mice.@*METHODS@#Except for normal mice, 32 HFD-induced obese mice were randomized into HFD, APS (1,000 mg/kg APS), BBR (200 mg/kg BBR), and APS plus BBR (1,000 mg/kg APS plus 200 mg/kg BBR) groups, respectively. After 6-week treatment (once daily by gavage), the obesity phenotype and pharmacodynamic effects were evaluated by histopathological examination of epididymal fat, liver, and colon using hematoxylin-eosin staining and serum biochemical analyses by an automated chemistry analyzer. The feces were collected at the 12 th week, and taxonomic and functional profiles of gut microbiota were analyzed by 16S ribosomal ribonucleic acid (16S rRNA) sequencing.@*RESULTS@#Compared with HFD group, the average body weight of APS plus BBR group was decreased (P<0.01), accompanied with the reduced fat accumulation, enhanced colonic integrity, insulin sensitivity and glucose homeostasis (P<0.05 or P<0.01). Importantly, APS combined with BBR treatment was more effective than APS or BBR alone in improving HFD-induced insulin resistance (P<0.05 or P<0.01). 16S rRNA sequence-based analysis of fecal samples demonstrated that APS combined with BBR treatment exhibited a better impact on HFD-induced gut microbiota dysbiosis, exclusively via the enriched abundances of Bacteroides, which corresponded to the large increase of predicted bacterial genes involved in carbohydrate metabolism.@*CONCLUSION@#APS combined with BBR may synergistically reduce obesity and modulate the gut microbiota in HFD-fed mice.

Research progress on Astragali Radix and prediction of its quality markers (Q-markers) / 中国中药杂志

Astragali Radix, a medicinal herb for invigorating Qi, has anti-aging, anti-tumor, immunoregulatory, blood sugar-and lipid-lowering, anti-fibrosis, anti-radiation and other pharmacological effects. This article reviewed the studies about the chemical components and pharmacological effects of Astragali Radix. According to the theory of quality markers(Q-markers) of Chinese medicinal materials, we predicted the Q-markers of Astragali Radix from traditional efficacy, chemical component validity, measurability, plant phylogeny, and pharmacokinetis. The results showed that total polysaccharides, flavonoids(e.g., calycosin-7-O-β-D-glucoside, formononetin, calycosin, quercetin, and ononin), and saponins(e.g., astragalosides Ⅱ, Ⅲ, and Ⅳ) can be taken as the main Q-markers. This review lays a foundation for regulating the quality research and standard establishment of Astragali Radix, and benefits the control and quality supervision of the production process of Astragali Radix and its related products.

Desulfovibrio vulgaris, a potent acetic acid-producing bacterium, increased by Astragalus polysaccharides to attenuate nonalcoholic fatty liver disease in mice / 中国药理学与毒理学杂志

OBJECTIVE Although the underlying mechanism is largely unknown, gut dysbiosis has emerged as a central initiator of obesity-related diseases including nonalcoholic fatty liver disease (NAFLD), type 2 diabetes and meta?bolic syndrome. The emerging evidence support the use of prebiotics like herb-derived polysaccharides for treating NAFLD by modulating gut microbiome. So, our study focused on the microbiota-dependent anti-NAFLD effect and the exact mechanisms of Astragalus polysaccharides (APS) extracted from Astragalus mongholicus Bunge in high-fat diet (HFD) fed mice. METHODS Co-housing experiment was used to assess the microbiota dependent anti-NAFLD effect of APS. Then, targeted metabolomics and metagenomics were adopted for determining short-chain fatty acids (SCFAs) and bacteria that were specifically enriched by APS. Further in vitro experiment was carried out to test the capacity of SCFAs-producing of identified bacterium. Finally, the anti-NAFLD efficacy of identified bacterium was tested in HFD fed mice. RESULTS Our results first demonstrated the anti-NAFLD effect of APS in HFD fed mice and the contribution of gut microbiota. Moreover, our results indicated that SCFAs, predominantly acetic acid were elevated in APS-supplemented mice and ex vivo experiment. Metagenomics revealed that D. vulgaris from Desulfovibrio genus was not only enriched by APS, but also a potent generator of acetic acid, which showed significant anti-NAFLD effects in HFD fed mice. In addition, D. vulgaris modulated the hepatic gene expression pattern of lipids metabolism, particularly suppressed hepatic fatty acid synthase (FASN) and CD36 protein expression. CONCLUSION APS enriched D. vulgaris is effective on attenuating hepatic steatosis possibly through producing acetic acid, and modulation on hepatic lipids metabolism in mice. Further studies are warranted to explore the long-term impacts of D. vulgaris on host metabolism and the underly?ing mechanism.

Effect of Astragalus Polysaccharide on Radiosensitivity and Epithelial-mesenchymal Transition of Human Nasopharyngeal Carcinoma CNE-1 Cells / 中国实验方剂学杂志

Objective:Astragalus polysaccharide (APS) was used in combination with ionizing radiation (IR) to investigate the mechanism of APS on the radiosensitivity of human nasopharyngeal narcinoma CNE-1 cells and the epithelial-mesenchymal transition (EMT). Method:Cell counting kit-8 (CCK-8) was used to detect the cytotoxicity of different concentrations of APS (0，6.25，12.5，25，50，100，200 g·L-1) on CNE-1 cells. Colony formation assay was used to calculate the survival fraction (survival fraction, SF) of CNE-1 cells treated with 12.5 g·L-1 APS combined with different radiation doses (0，2，4，6 Gy). The linear quadratic equation mathematical model (LQ) was used to draw the radiosensitivity curve according to SF value. Cell scratch and transwell chamber test were used to detect the migration and invasion ability of cells in each group. The apoptosis of cells in each group was detected by flow cytometry, Western blot was used to detect the expressions of EMT markers, apoptosis markers and protein kinase B/extracellular regulated protein kinases (Akt/ERK) pathway proteins in each group. Result:The results of colony formation assay and radiosensitivity curve showed that the combination of non-toxic dose of 12.5 g·L-1 APS and radiation dose of 4 Gy could significantly increase the radiosensitivity of CNE-1 cells. Compared with blank group and IR group, APS combined with IR could significantly inhibit the migration and invasion of CNE-1 cells (P<0.05), and increase the rate of apoptosis (P<0.05). In addition, compared with the blank group and the IR group, APS combined with IR could significantly down-regulate the expressions of N-cadherin, p-Akt and p-ERK, and significantly up-regulate the expressions of E-cadherin, Bax and Caspase-3 (P<0.05). Conclusion:APS combined with IR can inhibit the migration and invasion of CNE-1 cells, and increase the apoptosis induced by radiotherapy, which may be related to the inhibition of EMT and Akt/ERK pathway.

Astragalus polysaccharides suppressed cisplatin-resistance of colorectal cancer TH-29/DDPcells via regulating miR-20a/TGFBR2 axis / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of astragalus polysaccharides (APS) on proliferation, invasion, apoptosis and drugresistance of cisplatin-resistant colorectal cancer (CRC) HT-29/DDP cells through regulating miR-20a/TGFBR2 axis, and to explore the possible mechanism. Methods: Human CRC HT-29 cells and HT-29/DDP cells were used as non-drug resistant and resistant cell models, respectively; HT-29/DDP cells were randomly divided into four groups, including untreated (HT-29/DDP) group, APS treatment group, miR-20a mimics + APS group, and si-TGFBR2 + APS group. qPCR and Western blotting were applied to detect the expressions of miR-20a and TGFBR2 in HT-29/DDP cells treated with different concentrations ofAPS (0, 0.5, 1.0, 1.5 and 2.0 mg/ml). Subsequently, dual luciferase reporter gene assay was used to verify whether TGFBR2 was a target gene of miR-20a. In addition, CCK-8, Transwell andAnnexin V-FITC/PI double staining were applied to examine the effect ofAPS on proliferation, invasion and apoptosis of HT29/DDP cells. Furthermore, subcutaneous HT-29/DDP cell xenograft model was established on nude mice, and the effect ofAPS on the growth of transplanted tumor was observed. Results: APS significantly inhibited the proliferation of HT-29/DDP cells in a dose-dependent manner (P<0.01). Meanwhile, the expression of miR-20a was down-regulated in HT-29/DDP cells treated with APS, while the expression of TGFBR2 was significantly up-regulated (all P<0.01). Additionally, dual luciferase reporter gene assay result showed that TGFBR2 was a direct target of miR-20a in HT-29/DDP cells and its expression was suppressed. Furthermore, APS could enhance the drug sensitivity of HT-29/DDP cells through downregulating the inhibitory effect of miR-20a on TGFBR2 expression, thereby suppressed proliferation and invasion, and induced apoptosis of HT-29/DDP cells in vitro and in vivo. It was also found that this effect was related with the suppression of PCNA and Bcl-2 proteins and promotion of Bax and Caspase-3 proteins. Conclusion: APS reverses the resistance of HT-29/DDPcells to cisplatin by down-regulating the inhibitory effect of miR-20a on TGFBR2 expression.

Effect of Astragalus Polysaccharide on Sweet Taste Receptor Pathway in Intestine of Rat Model Induced by High-sugar and High-fat Diet / 中国实验方剂学杂志

Objective:To observe the effect of astragalus polysaccharide (APS) on taste receptor 1 member 2 (T1R2)/taste receptor 1 member 3 (T1R3) sweet taste receptor pathway in intestine of rat model induced by high-sugar and high-fat diet. Method:SD rats were randomly divided into normal group, high-sugar and high-fat group and astragalus polysaccharide group. Rats in high-sugar and high-fat group and astragalus polysaccharide groups were fed with high-sugar and high-fat diet for 16 weeks, while rats in astragalus polysaccharide group were fed with APS (0.7 g·kg-1, per day) for 8 weeks during this period. Serum samples were collected to determine the levels of fasting blood glucose, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). Intestinum tenue was collected to determine mRNA expressions of T1R2/T1R3, α-gustducin (Gα gust), transient receptor potential cation channel subfamily member 5 (TRPM5) and proglucagon (PG) gene by Real-time PCR, and protein expressions of T1R2, Gα gust and glucagon-like peptide-1 (GLP-1) protein by Western blot. Result:Rats in high-sugar and high-fat group had significantly higher levels of TC, TG and LDL-C, and lower HDL-C level in serum than those in normal group (Pα gust and PG genes in intestine, were significantly down-regulated in high-sugar and high-fat group (PPα gust, TRPM5 and PG genes in intestine were significantly up-regulated in astragalus polysaccharide group (Pα gust and GLP-1 protein expressions was consistent with that of T1R2, Gα gust and GLP-1 mRNA expressions. Protein expressions of T1R2, Gα gust and GLP-1 and mRNA expression of T1R3 were significantly lower in astragalus polysaccharide group than those of control group (PConclusion:APS could improve disturbance of lipid metabolism and impairment of intestinal sweet taste receptor pathway for rat model induced by high-sugar and high-fat diet.

Inhibitory effect of astragalus polysaccharides on pulmonary inflammation in asthma model mice and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the inhibitory effects of astragalus polysaccharides (A P S) with different molecular weights on the pulmonary inflammaton in the asthmatic mice, and to elucidate their mechanisms. Methods: A total of 30 female BALB/c mice were selected. They were randomly divided into normal control group, asthma model group (model group), low molecular weight APS (APS-low) group, midde molecular wieght APS (APS-middle) and high molecular weight APS (APS-high) group, with 6 mice in each group. The mouse models of asthma in model group and APS groups were established by injecting and inhaling ovalbumin (OVA). The mice in APS-low, APS-middle and APS- high groups were separately given 0. 1 mL of 4 500, 15 000, 30 000 molecular weight APS for intrapeitoneal injecton before inhaled OVA The mice in normal control group were treated with the same amount of normal saline instead of OVA injected and inhalled. The behavioral changes of the mice were observed during the atomization with OVA, and the total number of WBC and the counts of inflammatory cells in the bronchoalveolar lavage fluid (BALF) were observed with light microscope; the pathological changes of lung tissue was observed by HE staining. Cytometric Bead Array (CBA) method was used to detect the levels of IL-4 and IFN -y in the serum as well as BALF. The spleen CD4 T cells were extracted and cultured, and the ratios of T h l, Th2, Thl7 and Treg cells were detected by flow cytometry. CBA method was used to detect the levels of IL-4, IFN-y, IL-17, and IL-10 in the culture supernatant. Results: Compared with normal control group, the mice in model group showed asthma-like symptoms such as sneezing, snouting nose and shortness of breath, inflammatory infiltration of lung tissue, airway mucosal edema, smooth muscle thickening; the IL-4 levels in serum and BALF were increased (P < 0 . 05), and the IFN-y levels in serum and BALF were significantly decreased (P < 0 . 05); IL-4 and IL-17 levels in cell culture supernatant were significantly increased (P < 0 . 05), and the IFN-y and IL-10 levels were significantly decreased (P < 0 . 05); the ratios of Th2 and Thl7 cells were significantly increased (P < 0 . 05), and the ratios of T h l and Treg cells were significantly decreased (P < 0 . 05). Compared with model group, the asthmatic symptoms of the mice in APS groups, for example, scratching nose and mouth, shortness of breath, and irritability, were relieved to varying degrees; the inflammatory cell infiltration and wall thickening were significantly improved; the levels of IL-4 in serum and BALF were significantly reduced (P < 0 . 05), and the IFN -y levels were increased (P < 0 . 05); the levels of IL-4 and IL-17 in the culture supernatant were significantly reduced (P < 0 . 05) and the levels of IFN-y and IL-10 were increased (P < 0 . 05); the ratios of Th2 and Thl7 cells were significantly decreased (P < 0 . 05), and the ratos of T h l and Treg cells were increased (P < 0 . 05). Compared between three APS groups, the asthmatic symptoms and lung tissue inflammatory infiltration were lightened obviously in APS-low group, the IL-4 levels and the ratios of Th2 and Thl7 cells in serum and BALF were significantly decreased (P < 0. 05), and the IFN-y levels in serum and BALF and the ratios of Th2 and Thl7 cells were significantly increased (P < 0 . 05). Conclusion: APS can significantly improve the situaton of airway inflammaton infiltraton and the symptoms of the asthmatic mice. APS plays a therapeutic role for asthma by regulating the Th 1 / Th 2 and Th l7 / Treg cell balance, decreasing the levels of IL-4 and IL-17 and increasing the levels of IFN -y and IL-10 level; the low molecular weight APS shows an obvious effect.

Research approach on quality control of Astragalus polysaccharides based on polysaccharide receptor theory / 中草药

Astragali Radix is a commonly used staple Chinese herbal medicine with high medicinal and edible value. Its chemical composition mainly contains saponins, flavonoids, polysaccharides, and amino acids. Among them, Astragalus polysaccharides (APS), as the main active component of Astragali Radix, failed to be used as a quality control index to evaluate germplasm resources due to its non-specificity and weaker controllability. The proposal and application of polysaccharide receptor theory provides a new way to solve the international problem of quality control of polysaccharide drugs. In this paper, several related research literatures on the quality control of polysaccharides from Astragali Radix and polysaccharide receptor theory in recent years were reviewed, and the research ideas of quality control of Astragalus oligosaccharide active center were proposed.

Relative molecular weight determination of astragalus polysaccharides for injection / 药学学报

To determine relative molecular weight of astragalus polysaccharides (APs), we used Shodex GS620 gel permeation chromatographic column and differential refraction detector (GPC-RI) with dextran as a reference. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and GPC combined with multi-angle laser light scattering detection (GPC-MALLS) were also used．GPC-RI measure showed four peaks of APs, with the Mw of 1 380 000, 231 000, 18 000, and 480, respectively. Three main peaks were found by GPC-RI-MALLS with the Mw as 1 148 000, 180 000, and 43 000, respectively. Strong signals in 155 000 and 18 000 were detected by MALDI-TOF-MS, which also indicated the sugar moieties of the APs as hexoses. From our study, we found that the GPC-RI method with universal correction is most suitable for Mw determination of the APs. Nevertheless, the three methods should be combined and contrasted with each other to obtain accurate information in research of polysaccharides from Chinese medicine.

Astragalus polysaccharides decrease proliferation, migration, and invasion but increase apoptosis of human osteosarcoma cells by up-regulation of microRNA-133a

Osteosarcoma (OS) has a high incidence, malignity, and frequency of recurrence and metastasis. In this study, we aimed to explore the potential anti-cancer effects of Astragalus polysaccharides (APS) on human OS MG63 cells as well as underlying mechanisms. Viability of MG63 cells was assessed by CCK-8 assay to determine the adequate concentration of APS. Then, effects of APS on MG63 cell proliferation, cell cycle distribution, apoptosis, and migration and invasion were analyzed by BrdU incorporation, PI staining, flow cytometry, and transwell assays, respectively. The expression levels of proteins involved in these physiological processes were assessed by western blot analysis. Afterwards, miR-133a level in APS-treated cells was determined by qRT-PCR, and whether APS affected MG63 cells through regulation of miR-133a was determined. Finally, the activation of c-Jun N-terminal protein kinase (JNK) pathway was detected. We found that APS treatment suppressed the viability, proliferation, migration, and invasion of MG63 cells, as well as induced cell apoptosis. Moreover, APS enhanced the expression of miR-133a in MG63 cells. Knockdown of miR-133a reversed the APS treatment-induced MG63 cell proliferation, migration and invasion inhibition, as well as cell apoptosis. Furthermore, APS inactivated JNK pathway in MG63 cells. Knockdown of miR-133a reversed the APS treatment-induced inactivation of JNK pathway in MG63 cells. To conclude, APS repressed proliferation, migration, and invasion while induced apoptosis of OS MG63 cells by up-regulating miR-133a and then inactivating JNK pathway.

Effect of astragalus polysaccharides on apoptosis,transdifferentiation and ROS content in renal tubular epithelial cells of diabetic nephropathy / 中国免疫学杂志

Objective:To investigate the effect of Astragalus polysaccharides on apoptosis,transdifferentiation and ROS content in renal tubular epithelial cells of diabetic nephropathy.Methods:HK-2 cells were divided into low glucose group,high glucose group and astragalus polysaccharide+high glucose group,cell proliferation was detected by CCK-8 assay after 48 h;cell apoptosis and ROS content was detected by flow cytometry;the expression of E-cadherin,α-SMA,STAT1,STAT3,p-STAT1,p-STAT3 protein were detected by Western blot.Results:The survival rate of cells in high glucose group was significantly lower than low sugar group (P<0.01),cell apoptosis rate,ROS content and E-cadherin,α-SMA,p-STAT1 and p-STAT3 protein expression was significantly higher than low sugar group (P<0.01),the cell survival rate in high glucose+astragalus polysaccharide group was significantly higher than high glucose group,cell apoptosis rate,ROS content and E-cadherin,α-SMA,p-STAT1 and p-STAT3 protein expression was significantly lower than high glucose group (P<0.01).Conclusion:Astragalus polysaccharides can promote the proliferation of renal tubular epithelial cells induced by high glucose,inhibit apoptosis and transdifferentiation,and the mechanism is related to down regulation of JAK/STAT signaling pathway.

Experimental study on effects of astragalus polysaccharide on treating EAE in mice and regulating activation of BV-2 microglial cell line / 中国免疫学杂志

Objective:To investigate the therapeutic effects of astragalus polysaccharide (APS) on experimental autoimmune encephalomyelitis(EAE) and to explore the regulating effects on microglia activation that is associated with the pathogenesis of EAE and its possible mechanisms.Methods:Animal experiments:EAE model was induced by MOG35-55in C57BL/6 mice.APS was given by gavage.EAE was scored according to a 0-5 scale to observe the therapeutic effects of APS.Cell experiments:The effects of lipopolysac-charide (LPS) on cell viability of BV-2 microglial cell line were investigated by MTT assay and then the appropriate concentration of LPS to activate the BV-2 microglial cell line was selected.The microglia activation model was established.The changes in BV-2 microglial cell line morphology were observed with an inverted microscope.The cytokines of TNF-α and IFN-γ in the cell culture supernatant of BV-2 microglial cell line were detected by ELISA.The activated BV-2 microglial cells were treated with APS in different concentrations.The regulatory roles of the APS on the BV-2 microglial cell activation were observed.Western blot and Real-time PCR method were used to measure the protein and mRNA level of the PD-L1 on the cell surface of BV-2 microglial cells treated with APS.Results:APS could effectively ameliorate the symptoms in EAE mice and could suppress neuroinflammation of EAE significantly.The microglia activation model in vitro induced by LPS was successful.APS in certain concentration could inhibit the activation of microglia,increase the viabilily of the active microglia.Meanwhile,it could downregulated the level of the cytokines including IFN-γ and TNF-α and upregulated the expression of protein and mRNA of PD-L1 on activated microglia.Conclusion:APS can effectively inhibit the autoimmune reaction of EAE and effectively suppress the microglia activation induced by LPS,reduce the pro-duction of IFN-γ and TNF-α.APS plays a crucial role in reducing the inflammation induced by microglia activation.The potential mech-anisms might be related to the upregualtion of the PD-1/PD-L1 pathway.

Astragalus polysaccharide attenuates liver injury in rats with chronic cadmium exposure / 基础医学与临床

Objective To explore the effects of Astragalus polysaccharides on liver injury induced by cadmium chloride. Methods The cadmium chloride solution was administrated i.v. to develope rat model of liver injury. Rats were randomized divided into three groups;control group, model group, Astragalus polysaccharides intervention group. After the 5 weeks, the rats were sacrificed and the livers were weighted, alanine aminotrans ferase(ALT), aspartate aminotransferase (AST) and lactate dehydrogenase(LDH), the contents of cadmium(Cd) was checked by plasma mass spectrometerthe(ICP-MS), the contents of interleukin-2(IL-2) and transforming growth factor-β1(TGF-β1) was measured by ELISA, the the protein expression of Bcl-2 and Bax was observed by immunohisto-chemistry staining method. Results The liver weight was increased in model group, the level of ALT, AST and LDH were also ascended in model group, the contents of Cd, TGF-β1 and the protein expression of Bax all increased in mode group(P＜0.05 or P＜0.01). The content of IL-2 and the protein expression of Bcl-2 were decreased in mode group (P＜0.05 or P＜0.01). Compared with the model group, the liver index was decreased in Astragalus polysaccharides intervention group, the levels of ALT, AST and LDH were also reduced in Astragalus poly-saccharides intervention group, the contents of Cd, TGF-β1 and the protein expression of Bax were all dropped in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01).The content of IL-2 and the protein expression of Bcl-2 increased in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01). Conclusions Astragalus polysaccharides may reduce the oxidative stress injury of rats liver cells indued by cadmium chloride, and its mechanism may be explained by regulation of Bcl- 2/Bax protein expression.

Astragalus polysaccharides downregulates apoptosis in HepG2 cell through Wnt/β-catenin signaling pathway / 中草药

Objective To investigate the effects of Astragalus polysaccharides (APS) on HepG2 cell proliferation and apoptosis through Wnt/β-catenin signaling pathway. Methods Cell proliferation ability and cell viability was measured by MTT assay. Annexin V-FITC/PI stain and Caspase-3 activity assay were used to detect cell apoptosis. The changes of Wnt/β-catenin signaling pathway activity after treated by APS (100 and 200 mg/L) was measured by luciferase assay. The expression levels of β-catenin, c-myc, and Cyclin D1 were detected by qRT-PCR and Western blotting. Results As the concentration of APS increases, the cell viability of HepG2 cells decreased significantly (P < 0.05). Compared with control group, the cells apoptosis rates of in APS 100 and 200 mg/L groups were increased remarkably (P < 0.05); The relative activities of Caspase-3 in APS 100 and 200 mg/L groups increased significantly (P < 0.05); Meanwhile, the activity of luciferase in APS 100 and 200 mg/L groups decreased significantly (P < 0.05). The expression level of cleaved Caspase-3 was increased significantly. The mRNA 1evel of β-catenin, Cyclin D1, and c-myc in APS 100 and 200 mg/L groups were decreased significantly (P < 0.05), and the expression levels of Bcl-2, β-catenin, c-myc, and Cyclin D1 protein in APS 100 and 200 mg/L groups were decreased significantly (P < 0.05, 0.01). Conclusion APS promote the apoptosis of HepG2 cells through the down-regulation of Wnt/β-catenin signaling pathway, which leads to the inhibition of apoptosis-related gene Bcl-2 expression.

Study on standardization method of Astragalus polysaccharide reference substance / 中草药

Objective A variety of techniques were adopted to establish a standardization method of Astragalus polysaccharides (APs) reference substance. This reference substance was used for quality control of APs for injection. Methods The total sugar content was quantified by UV spectrophotometry ration. The content of monosaccharide was determined by HPLC at detective wavelength of 250 nm after derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP). The high performance size exclusion chromatography method was used for the determination of molecular weight and distribution. TLC was applied to check the free monosaccharide. Then the glycosidic linkage position and configuration were detected by using gas chromatography tandem mass spectrometry and nuclear magnetic resonance spectroscopy (1H-NMR). A number of experimental parameters were applied to control the quality of APs reference substance. Results The APs was composed of rhamnose (Rha), galactose acid (GalA), glucose (Glc), galactose (Gal), and arabinose (Arb), and Glc was the main component and free monosaccharide were not detected in APs. The result of molecular weight test showed that there were four chromatographic peaks in APs, of which the main peak (peak 3) MW was 12 531 and the peak area ratio was 83%. The amount of sugar was 94% by UV test in which the glc was calculated as reference substance. There were eight main types of glycosidic bonds detected by GC-MS. Among them, the 1,4-glucose linkage was the main one. The glycosidic bond confirmed by NMR was α configuration. Conclusion The standardization method of APs reference substance has been established in this study. The identification method and determination of sugar amount are accurate and reliable. In the further research, APs reference substance will be used in quality control of APs products.

Effect of Astragali Radix and its components on rats model of harmful fluid retention in upper jiao / 中草药

Objective: To study the effect of Astragali Radix and its components, which were astragalus polysaccharides, astragaloside IV and astragalus calycosin, on the rats model of harmful fluid retention in the upper warmer. Methods: Rats were randomized into the blank group (0.5% CMC Na), the model group (0.5% CMC Na), the water decoction of Astragali Radix group (5.40 g/kg), the astragalus polysaccharides group (1.41 g/kg), the astragaloside IV group (50 mg/kg), the astragalus calycosin group (30 mg/kg), and the positive control group with Qili Qiangxin Capsule (1 g/kg), ten rats in each group. In addition to the control group, the harmful fluid retention in the upper warmer rats model were established in other groups which were induced by combined intervention factors of SC isoproterenol in the scapular region and tracheal intubation. After two weeks of gavage administration, the changes of body weight (BW), heart weight index (HWI), left ventricular mass index (LVWI), lung weight index (LWI), left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), plasma creatine kinase (CK), the lung permeability index, the pulmonary alveolus pumpback amount, and the pulmonary dry wet ratio (W/D) in rats model were detected. Results: Compared with model group, the water decoction of Astragali Radix group and its components groups can improve the general condition of rats in varying degrees, BW increased significantly (P < 0.05) and the levels of HWI, LVWI and CK were all decreased in different degrees (P < 0.05 or < 0.01). In the astragalus polysaccharides group, astragaloside IV group and astragalus calycosin group, the levels of LVEF, LVFS and the pulmonary alveolus pumpback amount in rats were significantly increased (P < 0.05), and LWI, the lung permeability index and W/D were significantly decreased (P < 0.05). Conclusion: Astragali Radix and its components can improve the function of heart and lung in response to injury in rats model of harmful fluid retention in the upper warmer.

Effects of astragalus polysaccharide on cardiac dysfunction in rabbits with severe scald injury / 中华烧伤杂志

Objective@#To investigate the effects of astragalus polysaccharide (AP) on cardiac dysfunction in rabbits with severe scald injury.@*Methods@#Sixty-four New Zealand white rabbits were divided into pure scald group and AP group according to the random number table, with 32 rabbits in each group. Rabbits in the two groups were all inflicted with 30% total body surface area full-thickness scald on the back. Immediately after injury, rabbits in two groups were intraperitoneally injected with lactated Ringer′s solution once for antishock. Rabbits in AP group were intraperitoneally injected with 10 mL AP solution with the dosage of 200 mg/kg 10 min after injury and the following 6 days respectively, once a day. Rabbits in pure scald group were injected with 10 mL normal saline instead. Eight rabbits of each group were respectively selected before injury hour (BIH) 1 and on post injury day(PID) 1, 3, 7, and 14 to collect blood samples from ear marginal vein, and then sacrificed immediately to collect hearts at each time point post injury. The morphology of myocardium was observed after HE staining. The serum content of cardiac troponin I (cTnI) was detected by enzyme-linked immunosorbent assay (ELISA). The serum content of aspartate transaminase (AST), creatine kinase (CK), CK isoenzyme-MB (CK-MB), lactate dehydrogenase (LDH) was detected by fully automatic chemistry analyzer. The content of angiotensin Ⅱ (Ang Ⅱ) in serum and myocardium was detected with radioimmunoassay and the content of endothelin 1 (ET-1) in serum and myocardium was detected by ELISA. Another 8 normal rabbits were sacrificed to detect the content of Ang Ⅱ and ET-1 in the myocardium as the value of the two groups of scalded rabbits at BIH 1. The serum content of superoxide dismutase (SOD) and malondialdehyde (MDA) was detected by ELISA. The values of whole blood viscosity (ηb), reductive viscosity of whole blood (ηr), plasma viscosity (ηp), hematocrit (HCT), erythrocyte rigidity index (TK), erythrocyte aggregation index (EAI), and erythrocyte sedimentation rate (ESR) were detected by fully automatic hematology analyzer. Data were processed with analysis of variance of factorial design, independent sample t test, and Dunnett test.@*Results@#(1) Compared with those in pure scald group, the degrees of cardiomyocyte swelling, steatosis, necrosis and rupture of muscle fiber were significantly alleviated in rabbits of AP group on PID 1 and 3. There was no obvious increase in cell size, no breakage of muscle fiber or infiltration of inflammatory cells in myocardial interstitium on PID 7. The myocardial tissue structure and muscle fiber arrangement returned to normal condition on PID 14, with no interstitial fibroblast hyperplasia or excessive extra cellular matrix deposition. (2) Serum content of cTnI, CK, and LDH of rabbits in AP group was significantly lower than that in pure scald group on PID 1, 3, and 7 (with t values from 2.69 to 13.99, P<0.05 or P<0.01), while there was no significant difference between the two groups on PID 14 (with t values from -0.32 to 0.68, P values above 0.05). Serum content of AST and CK-MB of rabbits in AP group was significantly lower than that in pure scald group on PID 1 and 3 (with t values from 2.21 to 12.65, P<0.05 or P<0.01), while there was no significant difference between the two groups on PID 7 and 14 (with t values from 0.03 to 1.67, P values above 0.05). (3) Serum content of Ang Ⅱ of rabbits in AP group was significantly lower than that in pure scald group from PID 1 to 14 (with t values from 3.38 to 32.58, P values below 0.01). Serum content of ET-1 of rabbits in AP group was significantly lower than that in pure scald group from PID 3 to 14 (with t values from 3.54 to 11.02, P values below 0.01), while there was no obvious difference on PID 1 (t=0.39, P>0.05). Content of Ang Ⅱ and ET-1 in myocardial tissue of rabbits in AP group was significantly lower than that in pure scald group from PID 1 to 7 (with t values from 1.27 to 13.79, P values below 0.01), while there was no obvious difference on PID 14 (with t values respectively 0.07 and 0.81, P values above 0.05). (4) Serum content of SOD of rabbits in AP group was respectively (15.65±2.64), (14.67±0.74), and (8.43±0.56) ng/mL on PID 1, 3, and 7, which was significantly higher than (6.35±0.83), (2.62±0.75), and (2.84±0.41) ng/mL in pure scald group (with t values from -29.79 to -9.10, P values below 0.01); while there was no obvious difference on PID 14 [with (4.02±0.26) ng/mL in pure scald group and (4.11±0.52) ng/mL in AP group, t=-0.01, P>0.05]. Serum content of MDA of rabbits in AP group was respectively (1.31±0.61), (1.72±0.64), and (0.65±0.42) μmol /mL on PID 1, 3, and 7, which was significantly lower than (1.68±0.57), (2.34±0.79), and (1.06±0.32) μmol/mL in pure scald group (with t values from 1.63 to 3.16, P<0.05 or P<0.01), while there was no obvious difference on PID 14 [with (0.31±0.09) μmol/mL in pure scald group and (0.24±0.08) μmol/mL in AP group, t=2.11, P>0.05]. (5) Values of ηb1 and EAI of rabbits in AP group were significantly lower than those in pure scald group from PID 1 to 7 (with t values from 2.718 to 11.170, P<0.05 or P<0.01), while there were no obvious differences on PID 14 (with t values respectively 2.078 and -1.423, P values above 0.05). Values of ηb2 and ηr2 of rabbits in AP group were significantly lower than those in pure scald group on PID 3 and 7 (with t values from 2.178 to 19.205, P<0.05 or P<0.01), while there were no obvious differences on PID 1 and 14 (with t values from -0.730 to 1.320, P values above 0.05 ). Values of ηr1 and ESR of rabbits in AP group were significantly lower than those in pure scald group on PID 3, 7, and 14 (with t values from 3.021 to 8.058, P values below 0.01), while there were no obvious differences on PID 1 (with t values respectively 1.200 and 1.263, P values above 0.05 ). Value of ηp of rabbits in AP group was significantly lower than that in pure scald group on PID 1 (t=2.430, P<0.05), while there were no obvious differences on PID 3, 7, and 14 (with t values from 0.002 to 1.446, P values above 0.05 ). Value of HCT of rabbits in AP group was close to that in pure scald group on PID 1 (t=1.079, P>0.05), and the values were significantly lower than those in pure scald group on PID 3 and 14 (with t values respectively 3.849 and 4.208, P values below 0.01), while the value was significantly higher than that in pure scald group on PID 7 (t=-4.925, P<0.01). Value of TK of rabbits in AP group was lower than that in pure scald group on PID 7 (t=2.847, P<0.05), while there were no obvious differences on PID 1, 3, and 14 (with t values from -1.102 to 0.875, P values above 0.05).@*Conclusions@#AP can alleviate the damage of myocardium of rabbits with severe scald by reducing the production of vasoactive substances Ang Ⅱ and ET-1, decreasing oxidative stress injury by increasing the content of SOD and decreasing the production of MDA, improving blood flow performance and microcirculation perfusion.

Effect of astragalus polysaccharides on autophagy in nasopharyngeal carcinoma cell line CNE-2 / 实用医学杂志

Objective To investigate the effect of astragalus polysaccharides injection (APS) on autophagy of human nasopharyngeal cancer CNE-2 cells.Methods The inhibitory effect of APS on proliferation of CNE-2 cells was measured by CCK8 assay.Morphological changes of autophagy of APS was detected by acridine orange (AO) staining.Transmission electron microscopy was perform to observe the morphological alterations in the autophagic cells.The expression of Beclin-1 protein was detected by Western blot assay.Results APS markedly inhibited cell growth in a dose-and time-dependent manner.Increases of the number of large vacuoles and double layered membrane structure were observed by transmission electron microscopy.Results of AO staining revealed more bright red cytoplasm or nucleus in the ceradime group,which proved the existence of acidic vesicular organelles.Western blotting showed that Beclin-1 protein level was up-regulated.Conclusion APS may induce autophagy of human nasopharyngeal cancer CNE-2 cells via up-regulating the expression of apoptosis-related gene of beclin1 and inhibit the proliferation of CNE-2 cells.

Preparation and characterization of thermosensitive chitosan hydrogels containing astragalus polysaccharides/chitosan microshperes / 中国生化药物杂志

Objective To prepare thermosensitive chitosan (CTS) hydrogels containing astragalus polysaccharides (APS)/CTS microshperes (MS), and evaluate its physicochemical properties. Methods The APS/CTS MS (APS-MS) were prepared by spray drying method, and characterized by Scanning Electron Microscopy (SEM) and Laser Granularity Analyzer. Depending on the gelation temperature and gelating time, thermosensitive CTS hydrogels (HG) containing APS-MS (APS-MS-HG) were optimized by signal factor experiments, and the morphological characteristics were observed by SEM. In vitro release behaviors of APS-MS, hydrogels containing APS (APS-HG) and APS-MS-HG in pH 6.8 phosphate buffer were evaluated by dialysis tube method. Results The APS-MS were well dispersed with nearly spherical shapes and slightly wrinkled surfaces. The surface weighted mean D[3,2] of APS-MS was 8.078μm. The optimal APS-MS-HG, APS-MS-HG J, contained 3.012% APS-MS which were agitated with a magnetic stirrer for 3h. Observed by SEM, APS-MS were stayed spherical and dispersed unevenly in HG J, but the porous structure of HG J was disappeared in APS-MS-HG J. The release of APS from APS-MS-HG J was without initial burst release, and the cumulative amount of APS was about 74.75% after 36h. Conclusion Suppressing the phenomenon of sudden release at the first stage of delivery, APS-MS-HG J holds great promise for topical applications as a sustained-release nasal delivery system.