Astragalus membranaceus(Fisch.)Bge.administered by dissolving microneedles achieves systemic therapeutic effects at low doses Author links open overlay panel

Objective To determine the main components of Astragalus membranaceus(Fisch.)Bge(A.membranaceus,Huang Qi),Astragaloside IV(AIV)and Astragalus polysaccharides(AP),to characterize their properties,evaluate their in vivo efficacy,and to analyze drug diffusion using dissolving microneedle(DMN)technology in vivo.Methods Respectively,AIV-and AP-loaded DMNs comprising chitosan(CTS)and polyvinyl alcohol(PVA)were prepared via dual-mold forming.Their morphology,mechanical properties,in vivo solubility,and skin irritation characteristics were tested.In vivo efficacy was assessed in cyclophosphamide-induced immunosuppressed mice,in vivo diffusion of AIV and AP by DMNs and conventional methods was compared,and the rheological properties of AIV-CTS-PVA and AP-CTS-PVA mixtures were measured.Results Subcutaneous dissolution and absorption of AIV-CTS-PVA and AP-CTS-PVA microneedles(MNs)at low doses(50%–17%of intraperitoneal AIV injection and 12%–4%of intravenous AP injection)reduced the spleen index and acid phosphatase activity in immunosuppressed mouse models,increased the thymus index,and achieved equivalent or better systemic therapeutic effects.Compared with injections,AIV and AP achieved controllable solid-liquid conversion through delivery with CTS-PVA MNs,resulting in highly localized aggregation within 48 h,reducing the initial explosive effect of the drug,and achieving stable and slow drug release.Conclusion The present study enhances our understanding of the efficacy and remote effects of drug-loaded DMNs from a traditional Chinese medicine(TCM)perspective,thereby promoting the development of precise and efficient delivery of TCM and further expanding the drug-loading range and application scenarios for DMNs.

Recent efficacy and long-term survival of Astragalus polysaccharide combined with gemcitabine and S-1 in pancreatic cancer

BACKGROUND Pancreatic cancer is a highly malignant tumor with a rapid progression rate and a high susceptibility to infiltration and metastasis.Astragalus polysaccharide(APS),a pure Chinese medicine preparation primarily made from the traditional Chinese herb Astragalus,plays a positive role in the treatment of many malignant tumors.AIM To explore the recent efficacy of APS combined with gemcitabine plus tegafur gimeracil oteracil potassium capsule(S-1)(GS)regimen in the treatment of pancreatic cancer and assess its effect on the immune function and long-term survival of patients.METHODS A total of 97 patients who were diagnosed with pancreatic cancer and received GS chemotherapy at The First Affiliated Hospital of Zhejiang Chinese Medical University(Zhejiang Provincial Hospital of Chinese Medicine)from March 2021 to December 2021 were included in the retrospective analysis.Among them,41 patients received APS combined with GS chemotherapy,and 56 patients received GS chemotherapy only.The recent efficacy,immune function,adverse reactions,and long-term survival were compared among these patients.RESULTS After 4 cycles of treatment,the objective response rate of patients receiving the combined therapy of APS and GS was 51.22%,and the disease control rate(DCR)was 56.10%,higher than those of patients receiving the monotherapy with GS alone(30.36%and 35.71%,respectively).Besides,the percentages of CD3+T cells(50.18%±9.57%)and CD4+T cells(31.52%±5.33%)in the peripheral blood of patients receiving the combined therapy of APS and GS were higher compared with those treated with GS regimen alone[(44.06%±8.55%)and(26.01%±7.83%),respectively].Additionally,the incidences of leukopenia,thrombocytopenia,and fatigue in patients receiving the combined therapy of APS and GS were significantly lower than those in patients receiving the monotherapy of GS alone(17.07%,9.76%,31.71%vs 37.50%,28.57%,60.71%).Moreover,the median survival time of patients receiving the combined therapy of APS and GS was 394 days,significantly longer than that of patients receiving the mono-therapy of GS alone(339 days)(hazard ratio:0.66;95%CI:0.45-0.99;P=0.036).All these differences were statistically si-gnificant(P<0.05).CONCLUSION The combined therapy of APS and GS improved the recent efficacy and long-term survival of patients with pancreatic cancer and alleviated chemotherapy-induced immune suppression and adverse reactions.

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.

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.

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.

Astragalus polysaccharides can regulate cytokine and P-glycoprotein expression in H22 tumor-bearing mice

AIM:To investigate the adjunct anticancer effect of Astragalus polysaccharides in H22 tumor-bearing mice.METHODS:To establish a solid tumor model,5.0 × 10 6 /mL H22 hepatoma cells were inoculated subcutaneously into the right armpit region of Kunming mice(6-12 wk old,18-22 g).When the tumors reached a size of 100 mm 3,the animals were treated as indicated,and the mice were randomly assigned to seven groups(n = 10 each).After ten days of treatment,blood samples were collected from mouse eyes,and serum was harvested by centrifugation.Mice were sacrificed,and the whole body,tumor,spleen and thymus were weighed immediately.The rate of tumor inhibition and organ indexes were calculated.The expression levels of serum cytokines,P-glycoprotein(P-GP) and multidrug resistance(MDR) 1 mRNA in tumor tissues were detected using enzyme-linked immunosorbent assay,Western blotting,and quantitative myeloid-derived suppressor cells reverse transcription-polymerase chain reaction,respectively.RESULTS:The tumor inhibition rates in the treatment groups of Adriamycin(ADM) + Astragalus polysaccharides(APS)(50 mg/kg),ADM + APS(100 mg/kg),and ADM + APS(200 mg/kg) were significantly higher than in the ADM group(72.88% vs 60.36%,P = 0.013;73.40% vs 60.36%,P = 0.010;77.57% vs 60.36%,P = 0.001).The spleen indexes of the above groups were also significantly higher than in the ADM group(0.65 ± 0.22 vs 0.39 ± 0.17,P = 0.023;0.62 ± 0.34 vs 0.39 ± 0.17,P = 0.022;0.67 ± 0.20 vs 0.39 ± 0.17,P = 0.012),and the thymus indexes of the ADM + APS(100 mg/kg) and ADM + APS(200 mg/kg) groups were significantly higher than in the ADM group(0.20 ± 0.06 vs 0.13 ± 0.04,P = 0.029;0.47 ± 0.12 vs 0.13 ± 0.04,P = 0.000).APS was found to exert a synergistic antitumor effect with ADM and to alleviate the decrease in the sizes of the spleen and thymus induced by AMD.The expression of interleukin-1α(IL-1α),IL-2,IL-6,and tumor necrosis factor-α(TNF-α) was significantly higher in the ADM + APS(50 mg/kg),ADM + APS(100 mg/kg) and ADM + APS(200 mg/kg) groups than in the ADM group;and IL-10 was significantly lower in the above groups than in the ADM group.APS could increase IL-1α,IL-2,IL-6,and TNF-α expression and decrease IL-10 levels.Compared with the ADM group,APS treatment at a dose of 50-200 mg/kg could downregulate MDR1 mRNA expression in a dose-dependent manner(0.48 ± 0.13 vs 4.26 ± 1.51,P = 0.000;0.36 ± 0.03 vs 4.26 ± 1.51,P = 0.000;0.21 ± 0.04 vs 4.26 ± 1.51,P = 0.000).The expression level of P-GP was significantly lower in the ADM + APS(200 mg/kg) group than in the ADM group(137.35 ± 9.20 mg/kg vs 282.19 ± 20.54 mg/kg,P = 0.023).CONCLUSION:APS exerts a synergistic anti-tumor effect with ADM in H22 tumor-bearing mice.This may be related to its ability to enhance the expression of IL1α,IL-2,IL-6,and TNF-α,decrease IL-10,and downregulate MDR1 mRNA and P-GP expression levels.

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.

The effects of <i>astragalus</i>polysaccharide on zebrafish cell apoptosis and senescence

Astragalus polysaccharide (AP) is the extraction of astragalus, which is a plant used in traditional Chinese herb medicine and may increase an orgainism’s resistance to stress. Several earlier studies in vitro have indicated that AP has anti-aging activities, however the mechanism underlyling these activities was unclear and remained to be elucidated. In this study, Using the zebrafish (Danio rerio), we evaluated molecular mechanism of the effect of AP on zebrafish growth, development and apoptosis. 30 zebrafish embryos (24 hours post fertilization (hpf)) were exposed to varying concentrations of AP (from 0.125 mg/ml to 0. 5 mg/ml) continuously for 3 days. The results of β-galactosidase (SA-β-gal) and acridine orange fluorescence showed that AP can delay zebrafish embryos apoptosis under the concentration of 0.125 mg/ml. In addition, the differential gene expression of AP treated zebrafish embryos was examined by RT-PCR analysis. We found that the gene expression of mdm2 and tert were up-regulated while bax, p21 and p53 gene expression were down-regulated during early apoptosis of the zebrafish embryos mediated by AP. These results demonstrated that AP may play a role during the induction of senescence and this function might by p53-mediated pathway.

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.

Effects of astragalus polysaccharide on cell injury and mitochondrial pathway apoptosis in the hypoxia reoxygenation of myocardial cells

Objective: To study the effects of astragalus polysaccharide on cell injury and mitochondrial pathway apoptosis in the hypoxia reoxygenation of myocardial cells. Methods: H9c2 myocardial cell lines were cultured and divided into negative control group (NC group), hypoxia reoxygenation group (H/R group) and astragalus polysaccharide group (APS), H/R group underwent hypoxia reoxygenation treatment, and APS group underwent both hypoxia reoxygenation treatment and astragalus polysaccharides intervention. The cell viability was measured 8 h, 16 h and 24 h after reoxygenation;the expression of mitochondrial apoptosis genes, apoptosis pathway genes as well as the contents of ROS metabolism indexes were determined 24 h after reoxygenation. Results: 8 h, 16 h and 24 h after reoxygenation, the cell viability of H/R group were lower than those of NC group, and the cell viability of APS group were higher than those of H/R group;24 h after reoxygenation, BIM, BAX, Caspase-9, Caspase-3, p-PI3K, p-AKT and p-FoxO3a protein expression as well as ROS, HSP70 and p-p38MAPK contents in H/R group were significantly higher than those in NC group whereas BCL2 protein expression and SOD content were significantly lower than those in NC group;BIM, BAX, Caspase-9, Caspase-3, p-PI3K, p-AKT and p-FoxO3a protein expression as well as ROS, HSP70 and p-p38MAPK contents in APS group were significantly lower than those in H/R group whereas BCL2 protein expression and SOD content were significantly higher than those in H/R group. Conclusion: Astragalus polysaccharide can reduce the cell damage and inhibit the mitochondrial pathway apoptosis in the hypoxia reoxygenation process of myocardial cells.

Astragalus polysaccharide attenuates rat experimental colitis by inducing regulatory T cells in intestinal Peyer's patches

AIM: To explore probable mechanism underlying the therapeutic effect of Astragalus polysaccharide(APS) against experimental colitis.METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups. Colitis was induced with 2, 4, 6-trinitrobenzene sulfonic acid(TNBS). The rats with colitis were treated with 400 mg/kg of APS for 7 d. The therapeutic effect was evaluated by colonic weight, weight index of the colon, colonic length, and macroscopic and histological scores. The levels of regulatory T(Treg) cells in Peyer's patches were measured by flow cytometry, and cytokines in colonic tissue homogenates were analyzed using enzyme-linked immunosorbent assay. The expression of related orphan receptor-gt(ROR-gt), IL-23 and STAT-5a was measured by Western blot.RESULTS: After 7-d treatment with APS, the weight index of the colon, colonic weight, macroscopical and histological scores were decreased, while the colonic length was increased compared with the model group. The expression of interleukin(IL)-2, IL-6, IL-17, IL-23 and ROR-gt in the colonic tissues was down-regulated, but Treg cells in Peyer's patches, TGF-β and STAT5 a in the colonic tissues were up-regulated.CONCLUSION: APS effectively ameliorates TNBSinduced experimental colitis in rats, probably through restoring the number of Treg cells, and inhibiting IL-17 levels in Peyer's patches.

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.

Influence of astragalus polysaccharide on kidney status and fibrosis indices of a ratmodel of streptozotocin-induced diabetic nephropathy

Object： To examine the effect of astragalus polysaccharide （APS） on kidney status and fibrosis indices of rats withdiabetic nephropathy. Methods： 72 male rats were randomly divided into three groups： negative control group （NC, n =24）; diabetic nephropathy model group （DNM, n = 24）; and diabetic nephropathy model with APS group （DNM ＋ APS,n = 24）. Rats of the DNM and DNM ＋ APS groups were subjected to both unilateral nephrectomy and administeredstreptozotocin （STZ） injection （65 mg/kg）. DNM ＋ APS group rats were administered 50 IU/kg/d APS by subcutaneousinjection from the first week after operation until death. The NC and DNM group rats were subcutaneously injected withan identical volume of physiological saline. At weeks 3, 8, and 13 after the operation, 6 rats from each group wererandomly sacrificed and blood was collected to measure serum creatinine and blood urea nitrogen. On the day beforesacrifice, the rats were placed in a metabolic cage for 24 h to collect urine. At week 14 after the operation, 6 rats fromeach group were randomly selected to measure body weight and kidney index. Blood was collected to measure bloodglucose. The kidneys were harvested to detect pathological changes by hematoxylin and eosin staining. Results：Histological assessment of DNM rats suggested damage symptoms as evidenced by hyperplasia of the glomerularmesangial matrix, atrophia of the kidney tubules, and thickening of the basement membrane. In contrast, STZ-induceddiabetic nephropathy rats treated with APS （50 IU/kg/d） showed significantly improved histological results, suggestingthat APS has beneficial effect on renal tissues in STZ-induced DNM rats. Our results also indicated that APS relievedrenal injury and effectively improved body weight in DNM rats. The ratio of kidney weight to body weight was reducedand the early stage of renal function damage was improved after APS treatment. In the later stages of the disease, the 24h urinary protein significantly decreased. Moreover, APS down-regulated TGF-β1 and α-SMA expression of the kidney.Conclusion： APS significantly improved renal tubular interstitial injury in DNM rats and the early stage of renalfunction damage. The mechanism may be related to downregulation of the expression of TGF-β1 and α-SMA whichdelays the progression of renal interstitial fibrosis in DNM rats.

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.

黄芪多糖在Her-2阳性乳腺癌患者化疗中增效减毒的作用及其对血清Her-2-ECD、TAP和疗效的影响

目的:探讨黄芪多糖在人表皮生长因子受体-2(human epidermal growth factor receptor-2,Her-2)阳性乳腺癌患者化疗中增效减毒的作用及其对血清人表皮生长因子受体-2胞外段(human epidermal growth factor receptor-2-extracellular domain,Her-2-ECD)、肿瘤异常糖链糖蛋白(tumor abnormal protein,TAP)和疗效的影响。方法:选择2020年1月—2023年6月新余北湖医院收治的90例Her-2阳性乳腺癌患者作为研究对象,将患者随机分为对照组和观察组,各45例。两组均给予TCbHP(多西他赛+卡铂+曲妥珠单抗+帕妥珠单抗)方案治疗;观察组在此基础上给予注射用黄芪多糖治疗。比较两组治疗前后的中医症候积分、血清Her-2-ECD、TAP水平、免疫功能(CD4^(+)、CD8^(+))、白细胞(white blood cell,WBC)水平、乳腺癌患者的生活质量量表(functional assessment of cancer therapy-breast,FACT-B)评分、疗效及不良反应发生率。结果:治疗前,两组各项中医症候积分、Her-2-ECD、TAP、CD4^(+)、CD8^(+)、WBC、FACT-B评分比较,差异均无统计学意义(P>0.05)。治疗后,观察组各项中医症候评分、Her-2-ECD、TAP、CD8^(+)、FACT-B各项评分及不良反应发生率均低于对照组,CD4^(+)、WBC、总有效率均高于对照组,差异均有统计学意义(P<0.05)。结论:黄芪多糖能够改善Her-2阳性乳腺癌化疗患者的化疗效果,减轻化疗带来的毒性反应,提高患者的生活质量。

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.

Effect of Astragalus Polysaccharide on the Cell-mediated Immunity of Traumatic Stress Mice

To investigate the changes of immune functions and the effects of Astragalus polysaccharide (ASP) on the cell-mediated immunity of the traumatic stress model of mouse by amputation, 50 mice were randomly divided into 5 groups for study, in which the group A and B served as the normal control (by injecton of 0.5 ml of saline intra-peritoneally daily), and as the stress control (by intra-peritoneal injecton of 0.5 ml of normal saline into mice after amputation) respectively, to the group C, D and E of mice, 1000 mg/kg (high dose), 300 mg/kg (median dose) and 250 mg/kg (low dose). The CD4 + and CD8 + T cells as well as the expression of the c-fos protein were determined by immunohistochemical techniques, and the expressions of NF-κB mRNA and IL-10 mRNA were assayed by hybridization in situ . The experimental results showed that in comparison with the normal control group of mice (group A), the expression levels of NF-κB mRNA, IL-10 mRNA and the c-fos protein in the tissues of thymus and spleen in the stress controls were significantly elevated and the CD4 + T cells and CD4/CD8 ratio were decreased. However, in comparison with the stress control of mice (group B), the expressions of NF-κB mRNA and IL-10 mRNA were inhibited by ASP, and the CD4 + T cells and CD4/CD8 ratio were increased in groups C, D and E, but the level of c-fos protein was decreased. There was no significant difference in these parameters among group C, D and E. It is concluded that the functions of cell-mediated immunity of mice were disturbed under the stress condition of the traumatic injuries after amputation. And the immune functions can be effectively restored by the use of Astragalus polysaccharide.