Extraction of Astragalus Polysaccharides and Ganoderma lucidum Mycelia Polysaccharides and Their in Vitro Antioxidative Effects

[Objectives]To explore the extraction and in vitro antioxidant effects of astragalus polysaccharides(APS)and Ganoderma lucidum mycelia polysaccharides(GLMPS).[Methods]By studying the polysaccharides of the herbal medicinal material Astragalus membranaceus and the fungal medicinal material Ganoderma lucidum mycelia,two polysaccharides were mixed according to different proportions and concentrations by using the principle of traditional Chinese medicine compound combination.The effect of polysaccharides on the scavenging ability of hydroxyl radical system was determined by salicylic acid method.[Results]When the compound ratios of GLMPS and APS were 1∶1,1∶4,1∶5,4∶1,and 5∶1,the scavenging effect of compound polysaccharides was better than that of single-component polysaccharides,and with the increase of concentration,the scavenging effect increased.When the ratio of GLMPS and APS was 5∶1,the hydroxyl radical scavenging rate of the compound polysaccharide reached 59.77%,which was 18.72%higher than that of single GLMPS and 28.58%higher than that of single APS.The scavenging effect of compound polysaccharide is closely related to the compound ratio and concentration.[Conclusions]APS and GLMPS can obtain better hydroxyl radical scavenging ability than single-component polysaccharides through compounding in appropriate proportions.In addition,within a suitable concentration range,as the concentration increases,the scavenging ability also increases.

Effect of Astragalus polysaccharides on Erythrocyte Immune Adherence of Chickens Inoculated with Infectious Bursal Disease Virus

Two hundred and forty specific pathogen free leghorn chickens were randomly divided into four groups and reared in isolated pens. The tested chickens were negative to infectious bursal disease virus （IBDV） at 25 d old. Group 1 was treated with saline, whereas Groups 2, 3, and 4 were inoculated with 0.3 mL IBDV suspension intranasally the next day. Groups 3 and 4 were also administered with Astragalus polysaccharides （APS） intramuscularly twice daily at 5 or 10 mg kg-1 BW, respectively, until 31 d old. The erythrocyte-C3b receptor rosette rate （E-C3bRR） and the erythrocyte-C3b immune complex rosette rate （E-ICRR） were measured at 25, 29, 32, 35, and 38 d old. The results showed that IBDV significantly reduced E-C3bRR and E-ICRR when compared with the control group （P 〈 0.05）, while simultaneous administration of APS with 1BDV maintained E-C3bRR at similar levels to the control group （P 〉 0.05） and increased E-ICRR when compared with the control group and the group non-treated with APS （P 〈 0.05）. APS treatment reduced the morbidity and mortality of chickens inoculated with IBDV （P 〈 0.05）. The results suggest that APS may enhance the immune adherence of chickens erythrocytes by affecting the activity and/or the number of complement receptors on the erythrocyte membrane. These findings can be beneficial in providing an understanding of the basic mechanisms required for the rational application of APS in modern medicine.

Synergistic protection of astragalus polysaccharides and matrine against ulcerative colitis and associated lung injury in rats

BACKGROUND Ulcerative colitis(UC)is a main form of inflammatory bowel disease.Due to complicated etiology and a high rate of recurrence,it is quite essential to elucidate the underlying mechanism of and search for effective therapeutic methods for UC.AIM To investigate the effects of astragalus polysaccharides(APS)combined with matrine on UC and associated lung injury.METHODS UC was induced in rats by colon mucosal tissue sensitization combined with trinitro-benzene-sulfonic acid-ethanol.Then,the effects of the treatments of salazopyrine,APS,matrine,and APS combined with matrine on histopathological changes of lung and colon tissues,disease activity index(DAI),colon mucosal damage index(CMDI),serum endotoxin(ET)level,serum diamine oxidase(DAO)activity,the contents of tumor necrosis factor-αand interleukin-1β,and the activities of myeloperoxidase,superoxide dismutase,and malondialdehyde in lung tissues,as well as the protein expression of zonula occludens(ZO)-1,Occludin,and trefoil factor 3(TFF3)were detected in UC rats.RESULTS The treatments of salazopyrine,APS,matrine,and APS combined with matrine reduced DAI scores and improved histopathological changes of colon and lung tissues,as well as decreased CMDI scores,ET levels,and DAO activities in UC rats.Moreover,in lung tissues,inflammatory response and oxidative stress injury were relieved after the treatments of salazopyrine,APS,matrine,and APS combined with matrine in UC rats.Furthermore,the expression of ZO-1,Occludin,and TFF3 in lung and colon tissues was increased after different treatments in UC rats.Notably,APS combined with matrine exerted a better protective effect against UC and lung injury compared with other treatments.CONCLUSION APS combined with matrine exert a synergistic protective effect against UC and lung injury,which might be associated with regulating TFF3 expression.

Desulfovibrio vulgaris,a potent acetic acid-producing bacterium,increased by Astragalus polysaccharides to attenuate nonalcoholic fatty liver disease in mice

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 metabolic 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 underlying mechanism.

Functional Metabolomics Reveals that Astragalus Polysaccharides Improve Lipids Metabolism through Microbial Metabolite 2-Hydroxybutyric Acid in Obese Mice

Polysaccharides are widely present in herbs with multiple activities,especially immunity regulation and metabolic benefits for metabolic disorders.However,the underlying mechanisms are not well under-stood.Functional metabolomics is increasingly used to investigate systemic effects on the host by iden-tifying metabolites with particular functions.This study explores the mechanisms underlying the metabolic benefits of Astragalus polysaccharides(APS)by adopting a functional metabolomics strategy.The effects of APS were determined in eight-week high-fat diet(HFD)-fed obese mice.Then,gas chromatography–time-of-flight mass spectrometry(GC–TOFMS)-based untargeted metabolomics was performed for an analysis of serum and liver tissues,and liquid chromatography–tandem mass spectrom-etry(LC–MS/MS)-based targeted metabolomics was performed.The potential functions of the metabo-lites were tested with in vitro and in vivo models of metabolic disorders.Our results first confirmed the metabolic benefits of APS in obese mice.Then,metabolomics analysis revealed that APS supplemen-tation reversed the HFD-induced metabolic changes,and identified 2-hydroxybutyric acid(2-HB)as a potential functional metabolite for APS activity that was significantly decreased by a HFD and reversed by APS.Further study indicated that 2-HB inhibited oleic acid(OA)-induced triglyceride(TG)accumula-tion.It was also found to stimulate the expression of proteins in lipid degradation in hepatocytes and TG lipolysis in 3T3-L1 cells.Moreover,it was found to reduce serum TG and regulate the proteins involved in lipid degradation in high-fat and high-sucrose(HFHS)-fed mice.In conclusion,our study demonstrates that the metabolic benefits of APS are at least partially due to 2-HB generation,which modulated lipid metabolism both in vitro and in vivo.Our results also highlight that functional metabolomics is practical for investigating the mechanism underlying the systemic benefits of plant polysaccharides.

Advances in anti-tumor mechanism and clinical application of Astragalus polysaccharides

Astragalus polysaccharide has the beneficial effect of Qi-deficiency,and it also has several anti-tumor mechanisms,including pharmacological actions and clinical functions.Through literature review,this work concluded anti-tumor mechanism and clinical application of astragalus polysaccharide,which was of considerable significance to expand and optimize its anti-tumor function and clinical application.

The related effects of astragalus polysaccharides on the improvement of bone marrow suppression and hematopoietic stem cells during chemotherapy in elderly patients with lung cancer

Objective:To investigate the effects of astragalus polysaccharides(APS)on bone marrow suppression and hematopoietic stem cells during chemotherapy in elderly patients with lung cancer.Methods:120 elderly patients with lung cancer treated in the first hospital of Xingtai city from January 2019 to early December 2019 were divided into the treatment group and the control group by the random number table method,all of whom received pemetrexed+carboplatin chemotherapy,and the treatment group was treated with APS at the same time.The efficacy was evaluated after 2 cycles of chemotherapy,bone marrow suppression was observed,and levels of TCM symptoms score,peripheral blood T lymphocyte subgroup index,L-selectin(CD62L)and macrophage differentiation antigen-1(Mac-1)were measured before and after 2 cycles of chemotherapy.Results:The response rate(RR)was 56.67%in the treatment group and 45.00%in the control group,with no statistically significant difference(P>0.05);The disease control rate(DCR)in the treatment group was 81.67%,which was significantly higher than 65.00%in the control group(P<0.05);The reduction degree of leukopenia in the treatment group was significantly lower than that in the control group(P<0.05);The treatment group had a platelet reduction of grade 1+2 at a rate of 40.00%,and hemoglobin reduction of grade 1+2 at a rate of 28.33%,which were significantly lower than the control group at 65.00%and 58.33%(P<0.05);Compared with those before chemotherapy,the total score of TCM symptoms,serum CD62L and Mac-1 levels in the two groups all decreased significantly after chemotherapy,and they were significantly lower in the treatment group than in the control group(P<0.05);After chemotherapy,CD3+,CD4+and CD4+/CD8+in the treatment group increased significantly and they were all higher in the treatment group than in the control group,while CD8+decreased significantly and was lower in the treatment group than in the control group(P<0.05).There was no statistically significant difference in T lymphocyte subsets before and after chemotherapy in the control group(P>0.05).Conclusion:Astragalus polysaccharide can improve the chemotherapy effect and improve the bone marrow suppression in elderly patients with lung cancer,which may be related to its obvious enhancement of immune function and decrease of CD62L and Mac-1 levels.

Effects of astragalus polysaccharides on ventricular remodeling and miRNA-21 in rats with acute myocardial infarction

Objective:To investigate the effect of Astragalus polysaccharides(APS)on myocardial remodeling and expression of miR-21 after myocardial infarction.Methods:Sixty SPF grade healthy male rats were divided into the sham operation group,the model group,astragalus polysaccharide low,medium and high dose groups and atorvastatin group randomly with 10 rats in each group.The left anterior descending coronary artery(LAD)was ligated to establish myocardial infarction model in rats,and the corresponding drug intervention was given for 4 weeks.The changes of myocardial morphology and collagen were observed by HE and Masson staining.The levels of IL-1β,IL-6,TNF-αand IL-10 were detected by ELISA.The mRNA expressions of miR-21,MMP2,TIMP-2,Col-I,and Col-III was detected by RT-PCR.The protein expressions of TLR4,MyD88 and NF-κB p65 were detected by Western blot.Results:Compared with the model group,APS could improve the pathological morphology of myocardial tissue,increase the level of IL-10 in myocardial tissue,reduce the staining area of collagen and the contents of IL-1β,IL-6 and TNF-α(P<0.05).At the same time,APS could decreased the expression of MMP2,Col-I and Col-ⅢmRNA and the ratio of MMP2/TIMP-2,and increased the expression of TIMP-2 mRNA and miR-21 significantly(P<0.05).Furthermore,APS could significantly reduce the expression of TLR4,p-NF-κB p65 and MyD88 protein in myocardial tissue of rats with myocardial infarction,and the differences were statistically significant when compared with the model group(P<0.05).Conclusion:APS can inhibit the activation of TLR4/MyD88/NF-κB signaling pathway by upregulating the expression of miR-21,which plays a therapeutic role in ventricular remodeling after acute myocardial infarction.

Anti-obesity and Gut Microbiota Modulation Effect of Astragalus Polysaccharides Combined with Berberine on High-Fat Diet-Fed Obese Mice

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.

Inhibiting effect of Astragalus polysaccharides on the functions of CD4＋CD25^highTreg cells in the tumor microenvironment of human hepatocellular carcinoma

Background Astragalus polysaccharides （APS）, the main active extract from Astragalus membranaceus （a traditional Chinese medicinal herb）, is associated with a variety of immunomodulatory activities. The purpose of the present study was to examine the effect of APS on the function of Treg cells in the tumor microenvironment of human hepatocellular carcinoma （HCC） and to identify the pharmacologic mechanism of APS responsible for the anti-chemotactic activity in CD4＋CD25^highTreq cells in tumor site of HCC.

Protective effects of emodin and astragalus polysaccharides on chronic hepatic injury in rats

Background Chinese medicine plays an important role in hepatoprotective treatment. This study was conducted to investigate the protective effects of emodin and astragalus polysaccharides （APS） in a rat model of chronic hepatic injury. Methods Chronic hepatic injury was induced by hypodermic injection of an olive oil solution containing 40% carbon tetrachlodde （CCI4） twice a week, in addition to a diet of 79.5% maizena, 20% fat, 0.5% cholesterol, and 10% alcohol in the drinking water ad libitum for 12 weeks. Meanwhile, the rats were exposed to different concentrations of emodin （40 mg·kg^-1·d^-1）, APS （200 mg·kg^-1·d^-1）, combination drug （emodin 40 mg·kg^-1·d^-1 combined with APS 200 mg·kg^-1·d^-1） and colchicine （0.1 mg·kg^-1·d^-1） in parallel by oral gavage （once a day for 12 weeks）. At the end of 12 weeks, blood serum and liver tissue were taken. Serum was collected to determine the levels of total bilirubin （TBIL）, alanine transaminase （ALT）, aspartate transaminose （AST）, and albumin （ALB）. Liver and spleen indexes were assayed, followed by the measurements of the liver associated enzyme superoxide dismutase （SOD） and malondialdehyde （MDA）. Histopathological changes were studied using optical microscopy. Results Splenohepatomegalia was alleviated and serum levels of TBIL and ALT were reduced in the groups treated with emodin and APS when compared to the control group. In addition, the ALB level in the APS and combination groups was higher. Similarly, the SOD activity of liver homogenates was significantly higher in the groups treated with emodin and APS, while administration of the herbal derivatives prevented the elevation in MDA levels. Histological analysis showed that the APS and combination groups significantly ameliorated the hepatic injury. Conclusions Co-administration of emodin and APS demonstrated a synergistic action in reducing ALT and restoring ALB in the serum from a rat model of chronic hepatic injury. Emodin and APS may ameliorate the CCI4-induced hepatic injury in rats by elevating antioxidant-enzyme activities and reducing lipid peroxidation.

Influence of Different Extraction and Purification Methods on Astragalus Polysaccharides and Pharmacological Evaluation

Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides(APS) as complementary therapeutic agents prepared by different extraction and purification techniques.Methods Components of APS prepared by different extraction and purification techniques were analyzed,and these APS were used for synergy and attenuation of chemotherapy,radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice,and also used for the regulation of immune function to immunosuppression mice.Results Experimental data were analyzed by means of statistical method to get pharmaco-result:A3(extracted by microwave assistance and purified by membrane separation)>A4(extracted by refluxing and purified by membrane separation)>A1(extracted by refluxing and no purification)≈A2(extracted by microwave assistance and no purification).There were no significant differences on pharmacodynamic action between A1 and A2.However,compared with A1 and A2,it was worth noting that A3 and A4 exhibited good pharmacodynamic action.Then A3-in and A4-in,the samples in dialyzer after dialysis,were separated and purified to get homogeneous APS,which were the principal constituents of APS in dialyzer,with the molecular weight(Mw) of 7669 and 14142 determined by HPGPC,respectively.The average Mw of APS outside of the dialyzer,A3-out was 3102 and A4-out 3256,which were the main compositions of A3 and A4,accounted for 79.63% and 53.92%,respectively.Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity.Refluxing,microwave assistance extractions,and membrane enrichment techniques bring different cases on Mw distribution,components and pharmacodynamic action,and obviously exhibit relationship among component,Mw distribution,and pharmacological action.

Therapeutic and Immunoregulatory Effect of GATA-Binding Protein-3/T-Box Expressed in T-Cells Ratio of Astragalus Polysaccharides on 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis in Rats

Objective： To analyze the immunological characteristics of 2,4,6-trinitrobenzene sulfonic acid（TNBS）-induced colitis model and examine the therapeutic effects and mechanisms of Astragalus polysaccharides（APS） treatment. Methods： Thirty-two male specific pathogen free Spragne-Dawley rats were randomly equally assigned to four groups： control, TNBS, APS and prednisone groups. Experimental colitis was induced by enema administration of TNBS. Then rats were treated with APS（0.5 g·kg^（-1）·day^（-1）, once daily） or prednisone（1.0 mg·kg^（-1）·day^（-1）, once daily） by gavage for 14 days. Macroscopic lesion and histological damage were determined, and activity of myeloperoxidase（MPO） was measured in the colonic tissues. Expressions of T-box expressed in T-cells（T-bet） and GATA-binding protein-3（GATA-3） were determined by immunohistochemistry analysis and western blot. Results： Both macroscopic lesion and histological colonic damage induced by TNBS were reduced by APS and prednisone treatment. These were accompanied by significant attenuation of MPO activity（P=0.03）. TNBS intervention enhanced the expression of both GATA-3 and T-bet, but the expression of T-bet was significantly enhanced than that of GATA-3, resulting in significant reduction of GATA-3/T-bet ratio（P=0.025）. APS administration enhanced the expression of T-bet（P=0.04） and GATA-3（P=0.019） in comparison to TNBS group, and resulting in an up-regulated GATA-3/T-bet ratio. Prednisone treatment inhibited both expressions; however it also resulted in up-regulation of the GATA-3/T-bet ratio. Conclusions： These results demonstrated that APS exerted a beneficial immune regulatory effect on experimental colitis. It promoted the expression of T helper cell 1（Th1） and T helper cell 2（Th2） specific transcription factors but ultimately favor a shift toward Th2 phenotype, suggesting that APS possessed therapeutic potential in experimental colitis.

Improving Effects of Astragalus Polysaccharides on Cardiac Function via Keap1/Nrf2-ARE Signal Pathway in Adjuvant Arthritis Rats

Objective To observe the effect of Astragalus polysaccharides（APS） on cardiac function and expression of Keap1/Nrf2-ARE signal pathway in adjuvant arthritis（AA） rats. Methods Forty-eight rats were randomly divided into normal control（NC） group, model control（MC） group, leflunomide（LEF） group, and APS group. The AA model rats were induced with Freund＇s complete adjuvant, then administration began from day 19 after modeling for 30 d. The paw swelling and arthritic index（AI）, the cardiac function indexes, the expression of Keap1, maf, Nrf2 m RNAs, and HO-1, r-GCS proteins in cardiac tissues were observed. Oxidation-related substances（SOD, MDA, ROS, and TAC） and cytokines（IL-10 and TNF-ɑ） were also determined. Results Compared with NC group, the paw swelling and AI were increased, and the body weight was decreased in MC group. HR, HI, LVSP, LVEDP, and levels of MDA, TAC, ROS, RNS, and TNF-ɑ in MC group were increased, while dp/dtmax and levels of GSH, TRX, and IL-10 were decreased. Compared with MC group, LEF group showed higher HR and ± dp/dtmax, and lower LVEDP. In APS group, the HR, LVSP, and LVEDP were decreased and ±dp/dtmax was increased. TNF-ɑ was decreased, TAC, ROS, MDA, and IL-10 were increased in both LEF and APS groups. APS and LEF groups also showed less MDA, TAC, ROS, and SOD, and the differences were statistically significant. Spearman correlation analysis showed that the cardiac function parameters were positively correlated with Keap1, anti-oxidant indicators, and anti-inflammation cytokines, and negatively correlated with Nrf2, oxidation indictors, pro-inflammation cytokines, and inflammation indictors.Conclusion APS can adjust the expression of Keap1/Nrf2-ARE signal pathway in AA rats, and can also improve their cardiac function. The mechanism may be involved in increasing myocardial anti-oxidant capacity, reducing oxidative stress, and inhibiting inflammation.

Effects of Astragalus polysaccharide on immune function in B16 melanoma mice

Objective:To examine the effects of Astragalus polysaccharide on immune function in B16 melanoma mice.Method:Forty male C57BL/6 mice were divided equally into a control group,model group,Astragalus polysaccharide low-dose group,and Astragalus polysaccharide high-dose group,with 10 mice per group.B16 cells were used to develop a mouse model of melanoma.After B16 engraftment,40 mg/kg and 80 mg/kg Astragalus polysaccharide was administered by gavage every day to the Astragalus polysaccharide low-dose group and Astragalus polysaccharide high-dose group,respectively.Splenic index,thymic index,tumor growth curves,and tumor inhibition rates were measured.Flow cytometry was used to measure proportions of peripheral blood T lymphocyte subsets.Hematoxylin and eosin staining was used to examine histopathological changes in tumors.Immunofluorescence double staining was used to identify myeloid-derived suppressor cells in tumor tissues.Results:In the Astragalus polysaccharide high-dose group,splenic and thymic indices were significantly increased and tumor growth was inhibited in melanoma mice.Flow cytometry demonstrated increased CD4^(+)and CD4^(+)/CD8^(+)T-cell ratios in the high-dose Astragalus polysaccharide group.HE staining demonstrated significantly decreased numbers of tumor cells among mice with melanoma in the high-dose Astragalus polysaccharide group.Immunofluorescence double staining demonstrated significantly decreased numbers of myeloid-derived suppressor cells in tumor tissues in the high-dose Astragalus polysaccharide group.Conclusion:Astragalus polysaccharide inhibits tumor growth,increases splenic and thymic indices,increases CD4^(+)and CD4^(+)/CD8^(+)T-cell ratios,and decreases myeloid-derived suppressor cell numbers in melanoma mice.Our results indicate Astragalus polysaccharide may enhance immune function resulting in inhibition of tumorigenesis and tumor progression.

Antitumor effect of combination treatment of Astragalus polysaccharide and PD-L1 antibody on mice with Lewis lung carcinoma

Objective:To investigate the antitumor effect of combination treatment of Astragalus polysaccharide and PD-L1 antibody on mice with Lewis lung carcinoma.Methods:Forty male C57BL/6 mice were selected and divided equally into Model group,PD-L1 group,APSgroup,and APS+PD-L1 group.Lewis lung carcinoma cells were used to establish the lung carcinoma mouse model.After successful modeling,the PD-L1 group was injected with 200μg of PD-L1 antibody intraperitoneally on day 0/4/8/12;the APS group was gavaged with 80 mg/kg of APS daily for 14 days;the APS+PD-L1 group was gavaged with 80 mg/kg of APS daily for 14 days,and in addition,200μg of PD-L1 antibody was injected intraperitoneally on day 0/4/8/12.The spleen and thymus indices of each group of mice were observed,to plot the tumor growth curve and calculate the tumor suppression rate.The ratio of T-lymphocyte subsets in peripheral blood was measured by flow cytometry;the level of T cell-related cytokines in peripheral blood was detected by ELISA;MTT assay was used to detect the tumor-killing function of spleen lymphocytes in vitro.Results:PD-L1,APS,and APS+PD-L1 groups significantly increased spleen and thymus indices and inhibited tumor growth in lung carcinoma mice;flow cytometry results showed that PD-L1,APS,and APS+PD-L1 groups increased CD4^(+)T-cell ratio and CD4^(+)/CD8^(+)T-cell ratio;ELISA results showed that PD-L1,APS,and APS+PD-L1 groups significantly increased T cell-associated cytokine levels;MTT results showed that PD-L1,APS,and APS+PD-L1 groups enhanced the tumor-killing function of splenic lymphocytes in vitro.Conclusions:Astragalus polysaccharide can inhibit tumor growth,increase spleen and thymus indices,increase CD4^(+)T-cell ratio and CD4^(+)/CD8^(+)T-cell ratio,aswell as improve T-cell-related cytokine levels and splenic lymphocyte tumor-killing function in vitro in a mouse model of lung carcinoma,essentially inhibiting tumorigenesis and progression.

Self-adjuvant Astragalus polysaccharide-based nanovaccines for enhanced tumor immunotherapy:a novel delivery system candidate for tumor vaccines

The study of tumor nanovaccines(NVs)has gained interest because they specifically recognize and eliminate tumor cells.However,the poor recognition and internalization by dendritic cells(DCs)and insufficient immunogenicity restricted the vaccine efficacy.Herein,we extracted two molecular-weight Astragalus polysaccharides(APS,12.19 k D;APSHMw,135.67 k D)from Radix Astragali and made them self-assemble with OVA257–264directly forming OVA/APS integrated nanocomplexes through the microfluidic method.The nanocomplexes were wrapped with a sheddable calcium phosphate layer to improve stability.APS in the formed nanocomplexes served as drug carriers and immune adjuvants for potent tumor immunotherapy.The optimal APS-NVs were approximately 160 nm with uniform size distribution and could remain stable in physiological saline solution.The FITC-OVA in APS-NVs could be effectively taken up by DCs,and APS-NVs could stimulate the maturation of DCs,improving the antigen cross-presentation efficiency in vitro.The possible mechanism was that APS can induce DC activation via multiple receptors such as dectin-1 and Toll-like receptors 2 and 4.Enhanced accumulation of APS-NVs both in draining and distal lymph nodes were observed following s.c.injection.Smaller APS-NVs could easily access the lymph nodes.Furthermore,APS-NVs could markedly promote antigen delivery efficiency to DCs and activate cytotoxic T cells.In addition,APS-NVs achieve a better antitumor effect in established B16-OVA melanoma tumors compared with the OVA+Alum treatment group.The antitumor mechanism correlated with the increase in cytotoxic T cells in the tumor region.Subsequently,the poor tumor inhibitory effect of APS-NVs on the nude mouse model of melanoma also confirmed the participation of antitumor adaptive immune response induced by NVs.Therefore,this study developed a promising APS-based tumor NV that is an efficient tumor immunotherapy without systemic side effects.

Effects of Compound Herbal Medicine on Serum Biochemical Indexes and Egg Nutritional Quality in Laying Hens

[ Objective] To investigate the effects of traditional Chinese medicine compound preparation on serum biochemical indexes and egg nutri- tional quality of laying hens. [Method] A total of 1 200 chickens at the age of 44 weeks were selected and randomly divided into five groups. After administration with different dosages of astragalus polysaccharides and berberine for 9 weeks, serum biochemical indexes were determined and nu- trient contents of egg were detected. [ ltesultl The contents of total protein, globulin, high-density lipoprotein cholesterol （HDL-C） and alkaline phosphatase （ALP） in all treatment groups were significantly （ P 〈 0.05 or 0.01 ） higher than those in the control group. The cholesterol contents of all treatment groups were significantly （ P 〈 0.05 or 0.01 ） lower than those of the control group. E Conclusion~ The optimal concentrations of as- tragalus polysacchaddes and berberine in basic diets are 200 and 30 mg/kg, respectively.

Therapeutic Efficacy of Chinese Herbal Compound Preparation on Viral Diseases in Poultry

[Objective] To develop a new antiviral agent with high and long efficacy, wide spectrum, low toxicity and low cost from traditional Chinese herbal medicine and their compound. [ Method] The ＂Quxie＂ （ i. e. eliminating pathogenic factors） drugs including Hedyotis diffusa and Loni- cera japonica and ＂Fuzheng＂ （ i. e. strengthening body resistance） drugs including Radix astragali and Glycyrrhiza uralensis were taken to form an antiviral herbal compound preparation. Taking the astragalus polysaccharide monomer as control, the experiment was constructed by using the agent to treat some naturally occurring viral diseases, including Newcastle disease （ND）, infectious bursal disease （IBD）, duck viral hepatitis （DVH）, avian influenza （Al） H9 subtype and avian swollen head syndrome. [Ressult] The effective rates of the agent for treatment of the above- mentioned diseases were 87%, 87%, 69%, 100% and 100%, respectively, which were higher than that of astragalus polysaccharide monomer. The cure rates for these diseases were 33%, 37%, 13%, 69% and 75%, respectively, which were significantly higher than that of astragalus polysacchadde monomer （ P 〈 0.01 ）. The rates of poor therapeutic efficacy were 15%, 21%, 24%, 6% and 7%, respectively, which were significantly lower than that of astragalus polysaccharide monomer （P 〈 0.01 ）. [ Condusion] The Chinese herbal compound preparation shows good therapeutic effects on some avian viral diseases, which can decrease mortality rate and improve production performance of poultry more greatly than astragalus polysaccharide monomer.

Modulating effects of Astragalus polysaccharide on immune disorders via gut microbiota and the TLR4/NF-κB pathway in rats with syndrome of dampness stagnancy due to spleen deficiency

The syndrome of dampness stagnancy due to spleen deficiency(DSSD)is relatively common globally.Although the pathogenesis of DSSD remains unclear,evidence has suggested that the gut microbiota might play a significant role.Radix Astragali,used as both medicine and food,exerts the effects of tonifying spleen and qi.Astragalus polysaccharide(APS)comprises a macromolecule substance extracted from the dried root of Radix Astragali,which has many pharmacological functions.However,whether APS mitigates the immune disorders underlying the DSSD syndrome via regulating gut microbiota and the relevant mechanism remains unknown.Here,we used DSSD rats induced by high-fat and low-protein(HFLP)diet plus exhaustive swimming,and found that APS of moderate molecular weight increased the body weight gain and immune organ indexes,decreased the levels of interleukin-1β(IL-1β),IL-6,and endotoxin,and suppressed the Toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)pathway.Moreover,a total of 27 critical genera were significantly enriched according to the linear discriminant analysis effect size(LEfSe).APS increased the diversity of the gut microbiota and changed its composition,such as reducing the relative abundance of Pseudoflavonifractor and Paraprevotella,and increasing that of Parasutterella,Parabacteroides,Clostridium XIVb,Oscillibacter,Butyricicoccus,and Dorea.APS also elevated the contents of short-chain fatty acids(SCFAs).Furthermore,the correlation analysis indicated that 12 critical bacteria were related to the body weight gain and immune organ indexes.In general,our study demonstrated that APS ameliorated the immune disorders in DSSD rats via modulating their gut microbiota,especially for some bacteria involving immune and inflammatory response and SCFA production,as well as the TLR4/NF-κB pathway.This study provides an insight into the function of APS as a unique potential prebiotic through exerting systemic activities in treating DSSD.