Polysaccharides from Angelica sinensis alleviate neuronal cell injury caused by oxidative stress

Angelica sinensis has antioxidative and neuroprotective effects. In the present study, we aimed to determine the neuroprotective effect of polysaccharides isolated from Angelica sinensis. In a pre-liminary experiment, Angelica sinensis polysaccharides not only protected PC12 neuronal cells from H202-induced cytotoxicity, but also reduced apoptosis and intracellular reactive oxygen species levels, and increased the mitochondrial membrane potential induced by H202 treatment. In a rat model of local cerebral ischemia, we further demonstrated that Angelica sinensis poly-saccharides enhanced the antioxidant activity in cerebral cortical neurons, increased the number of microvessels, and improved blood flow after ischemia. Our findings highlight the protective role of polysaccharides isolated from Angelica sinensis against nerve cell injury and impairment caused by oxidative stress.

Polysaccharide from Angelica Sinensis Suppresses Inflammation and Reverses Anemia in Complete Freund's Adjuvant-induced Rats

The antinlammatory and antianemic activities of Angelica sinensis polysaccharide(ASP)isolated from roots of Angelica sinensis(AS)was investigated in a complete Freund's adjuvant(CFA)-induced arthritic rat model.It was observed that serum iron(SI)and total iron binding capacity(TIBC)levels were elevated after 4-week oral administration of ASP.Red blood cell(RBC)count and hemoglobin(Hb)concentrations were ameliorated as well.Moreover,infammatory cytokines IL-6 and TNF-a were decreased strikingly in CFA-induced arthritic rats after treatment of ASP.Evidence also showed that ASP strongly inhibited hepcidin expression through the Janus kinase/signal transducers and activators of transcription(JAK2/STAT3)pathway.Furthermore,ASP exhibited reduced primary and secondary lesions in adjuvant arthritis,attenuating synovitis and inflammatory joint damage.Data presented in this article collectively indicated that ASP significantly decreased proinflammatory cytokines(TNF-a,IL-6),which might play a crucial role in the CFA-induced arthritic rats,and had a therapeutic effect on adjuvant arthritis in rats.Results of Western blot analysis indicated that ASP inhibited the activation of IL-6/JAK2/STAT3 signaling pathway in the CFA-induced arthritic rats.

Preparation and Identification of Angelica sinensis Polysaccharide-iron Complex

Angelica sinensis polysaccharide（ASP） was extracted from Angelica sinensis by boiling water. An Angelica sinensis polysaccharide-iron complex（APC） was prepared under the alkaline condition by adding a ferric chloride solution to the ASP solution. Then some identifiable properties of the complex were studied. The content of iron（ Ⅲ ） in the complex was determined with iodometry. The thermal property, the microscopic structure, the spectral characteristics, and N, C, H contents of the complex were examined by a variety of techniques including DSC, TEM, IR, NMR, and elemental analysis. The content of iron（ Ⅲ ） in the complex ranges from 10% to 40%. The DSC result shows that the melting point of the complex is about 450 ℃. The TEM result shows that the complex has an iron（ Ⅲ ） core（β-FeOOH core） linked by hydroxy and oxy bridges, with the polysaccharide chains attached to the surface of the core. The IR and NMR results also show that there is a β-FeOOH core in the complex. The elemental analysis shows that the contents of N, C, H in the complex are, respectively, lower than those of N, C, H in ASP. All our studies indicate that the APC consists of a β-FeOOH core surrounded by ASP.

Angelica sinensis polysaccharides ameliorate 5-flourouracil-induced bone marrow stromal cell proliferation inhibition via regulating Wnt/β-catenin signaling

Chemotherapy may cause cellular oxidative stress to bone marrow.Oxidative damage of bone marrow hematopoietic microenvironment is closely related to chronic myelosuppression after chemotherapeutic treatment.Angelica sinensis polysaccharides(ASP)are major effective ingredients of traditional Chinese medicine Angelica with multi-target anti-oxidative stress features.In the current study,we investigated the protective roles and mechanisms of ASP on chemotherapy-induced bone marrow stromal cell(BMSC)damage.The human bone marrow stromal cell line HS-5 cells were divided into control group,5-FU group,5-FU+ASP group,and 5-FU+LiCl group to investigate the mechanism of ASP to alleviate 5-FU-induced BMSC proliferation inhibition.The results showed that 5-FU inhibits the growth of HS-5 cells in a time and dose-dependent manner;however,ASP partially counteracted the 5-FU-induced decrease in cell viability,whereas Wnt signaling inhibitor Dkk1 antagonized the effect of ASP on HS-5 cells.ASP reversed the decrease in total cytoplasmicβ-catenin,p-GSK-3β,and CyclinD1 following 5-FU treatment and modulated nuclear expression ofβ-catenin,Lef-1,and C-myc proteins.Furthermore,ASP also enhanced the antioxidant capacity of cells and reduced 5-FU-induced oxidative stress,attenuated FoxO1 expression,thus weakened its downstream apoptosis-related proteins and G0/G1 checkpoint-associated p27^(Kip1) expression to alleviate 5-FU-induced apoptosis and to promote cell cycle progression.All the results above suggest that the protective role of ASP in 5-FU-treated BMSCs proliferation for the chemotherapy may be related to its activating Wnt/β-catenin signaling and keeping homeostasis betweenβ-catenin and FoxO1 under oxidative stress.The study provides a potential therapeutic strategy for alleviating chemotherapeutic damage on BMSCs.

Study on Extraction Technology for Polysaccharide from Blood-Supplementing Angelica Sinensis Decoction

Purpose:Optimize water-alcohol technology for extracting polysaccharide from blood-supplementing angelica sinensis decoction to improve the extraction rate.Method:Draw the standard curve of glucose reference substance and build the regression equation to calculate the polysaccharide content.Investigate the effect of water addition times,extraction duration,extraction times,ethanol concentration for ethanol precipitation and times of ethanol precipitation on the extraction rate of polysaccharide.Result Add 8 times the medicinal material quality of water,and perform 2 extractions for 120min each time;it shows that conducting 2 ethanol precipitations with 80%ethanol concentration results in the maximum polysaccharide content,indicating the best extraction condition.Conclusion:The experiment establishes an easy and convenient water-alcohol method for extracting polysaccharide from blood-supplementing angelica sinensis decoction,and lays a research foundation on pharmacodynamics of polysaccharide in blood-supplementing angelica sinensis decoction.

Mitochondrial membrane stabilization by Angelica sinensis polysaccharide in murine aplastic anemia

In order to investigate the mechanism of mitochondrial membrane stabilization by Angelica sinensis polysaccharide (ASP) in murine aplastic anemia (AA).ICR mice were randomly divided into control, AA and ASP-treated groups. The AA group mice were treated with 60Coγand intraperitoneal injections of cyclophosphamide and chloramphenicol. The control animals were treated with lead shielding irradiation and saline injection. The treated AA mice were fed with ASP for 2 wk. Mitochondrial ultrastructure of the bone marrow was observed by transmission electron microscopy, and the transmembrane potential of bone marrow-nucleated cells (BMNC)was examined by fluorescence spectrophotometry. The Cox and MDH contents of the medium were also studied in the three groups.The mitochondrial number and transmembrane potential of BMNC in the bone marrow decreased in the AA group as compared to the control group, but improved in the ASP-treated group as compared to the AA group. Complete mitochondrial cleavage in the ASP-treated group was significantly delayed (P<0.05) as compared to the AA group. We conclude that ASP might improve mitochondrial membrane stabilization, and suppress the downregulation of transmembrane potential and apoptosis of BMNC in AA.

当归多糖调节骨髓造血微环境治疗再生障碍性贫血的机制

背景:如何改善造血微环境是目前再生障碍性贫血治疗的热点问题。目的:结合GEO测序分析、网络药理学与实验验证当归多糖改善骨髓造血微环境治疗再生障碍性贫血的作用机制。方法:借助GEO数据库获得了再生障碍性贫血相关差异表达基因并进行GO和GSEA分析。结合文献与PubChem、SwissTargetPrediction、PharmMapper数据库获取当归多糖活性成分和靶点。取交集靶点后利用STRING和Cytoscape构建蛋白相互作用网络,并进行GO和KEGG富集分析。构建再生障碍性贫血小鼠模型,利用血细胞分析仪、流式细胞术、ELISA、免疫组化、免疫荧光染色、Western blot方法验证当归多糖对再生障碍性贫血的疗效与作用机制。结果与结论:(1)共筛选出834个差异表达基因,参与细胞发育、造血、髓系细胞分化等生物学过程;(2)检索得到当归多糖相关347个靶点并筛选出77个潜在治疗基因,其中VEGFA、EGLN1、BCL2、干扰素γ、白细胞介素2、白细胞介素4、白细胞介素6等血管生成、凋亡及免疫相关因子度值显著;(3)KEGG通路富集分析治疗靶点主要富集在Th17细胞分化、NK相关细胞毒性、细胞黏附因子等干扰素γ、白细胞介素2、白细胞介素4相关信号通路;(4)动物实验表明,当归多糖能够显著改善再生障碍性贫血小鼠骨髓造血,增加外周血细胞及骨髓单个核细胞计数,提高小鼠存活率;与模型组相比,当归多糖组小鼠Th1/Th2细胞比例显著下降(P<0.01),干扰素γ水平显著降低(P<0.01),白细胞介素4水平升高(P<0.05),VEGFA表达显著升高(P<0.01),EGLN1表达显著降低(P<0.01),骨髓单个核细胞凋亡率、活性氧水平显著降低(P<0.01),Cleaved Caspase-3蛋白表达显著增高(P<0.01),Bcl-2/Bax比值显著降低(P<0.01);(5)结果表明:当归多糖能够通过调节异常T细胞亚群,促进血管生成以改善造血微环境,并抑制骨髓单个核细胞凋亡,以改善再生障碍性贫血小鼠的造血功能。

当归多糖ASP3的甲基化分析

通过部分酸水解和甲基化分析,结合GC-MS测试手段,对当归多糖ASP3的糖链结构进行了研究.采用0.2mol/L三氟乙酸(Trifluoroacetic acid,TFA)对ASP3进行了部分酸水解,对水解前后的多糖组分进行甲基化分析.结果显示:ASP3的糖残基主要由D-GalpA,D-Galp,L-Araf和L-Rhap组成,主链由1,4-D-GalpA连接,形成"光滑区"半乳糖醛酸聚糖,由1,4-D-GalpA通过O-4位与1,2-;1,2,4-L-Rhap的O-2位交替连接形成含有较多分支的"毛发区"鼠李半乳糖醛酸聚糖.58.8%的Rhap残基发生O-4位取代(1,2,4-L-Rhap).由T-,1,5-,1,3,5-Araf和T-,1,3-,1,3,6-,1,4-,1,4,6-D-Galp聚合形成的阿拉伯半乳聚糖、半乳聚糖以及阿拉伯聚糖是ASP3侧链的主要组成,通过Rhap残基的O-4位与主链相连.

当归多糖ASP3及其水解产物的NMR光谱分析

对当归多糖ASP3的糖链结构进行分析.分别采用0.2mol/L三氟乙酸(Trifluoroacetic acid,TFA)和内切-α-(1→4)-聚半乳糖醛酸酶(EndoPG)对ASP3进行部分酸水解和酶水解,并对水解前后多糖组分的1D和2D NMR光谱特征进行分析.实验结果表明,ASP3是一种果胶多糖.GalpA和Rhap位于多糖分子的主链,由1→4-D-GalpA相连形成的"光滑区"(半乳糖醛酸聚糖)是其主要组成部分;由α-(1→4)-GalpA通过O4位与α-(1→2)-和α-(1→2,4)-Rhap的O2位交替连接所形成的重复单元[→4)-α-GalpA-(1→2)-α-Rhap-(1→]构成具有较高分支的"毛发区"(富含中性糖侧链的鼠李半乳糖醛酸聚糖).Galp和Araf是中性糖侧链的主要组成,通过Rhap残基的O4位与主链相连.非还原性末端T-β-Galp,β-(1→3)-,β-(1→3,6)-,β-(1→4)-,β-(1→4,6)-Galp聚合形成以β-(1→3,6)-Galp为分支点的β-(1→6)-半乳聚糖和以β-(1→4,6)-Galp为分支点的β-(1→4)-半乳聚糖.T-α-Araf,α-(1→5)-Araf和α-(1→3,5)-Araf聚合形成以α-(1→3,5)-Araf为分支点的α-(1→5)-阿拉伯聚糖.此外,由α-(1→5)-阿拉伯聚糖通过α-(1→3)连接与β-(1→6)-半乳聚糖末端聚合形成阿拉伯半乳聚糖.

当归多糖对结直肠癌细胞顺铂耐药性的影响及作用机制

目的 探讨当归多糖对结直肠癌细胞顺铂耐药性的影响及作用机制。方法 将人结直肠癌细胞HT-15、顺铂耐药细胞株HT-15/DDP仅使用30μmol/L浓度的顺铂干预设为对照组,使用对应质量浓度1.0、1.5、2.0 mg/mL的当归多糖干预12 h后再加入30μmol/L浓度顺铂设为当归多糖组。通过脂质体细胞转染技术分别将miR-10b-5p inhibitor、TGFBR2 mimics转染至HT-15/DDP细胞中,再加入2.0 mg/mL当归多糖干预48 h,然后将其分为当归多糖(2.0 mg/mL)+miR-10b-5p inhibitor组、当归多糖(2.0 mg/mL)+TGFBR2 mimics组。MTT法检测细胞增殖能力;Transwell小室试验检测细胞迁移能力;流式细胞术检测细胞凋亡能力。RT-PCR检测miR-10b-5p、TGFBR2 mRNA表达水平;Western blot检测TGFBR2蛋白表达水平。结果 单独顺铂干预的对照组HT-15/DDP细胞增殖率、细胞迁移个数明显高于HT-15细胞(P<0.01);经不同质量浓度当归多糖预处理12 h后,可明显增强HT-15/DDP细胞对顺铂的敏感性,HT-15/DDP细胞增殖率、细胞迁移个数明显降低(P<0.01)。单独顺铂干预的对照组HT-15/DDP细胞凋亡率明显低于HT-15细胞(P<0.01);经不同质量浓度当归多糖预处理12 h后,明显增强了HT-15/DDP细胞对顺铂的敏感性,HT-15/DDP细胞凋亡率明显升高(P<0.01)。HT-15/DDP细胞miR-10b-5p、TGFBR2表达水平在各组间比较差异均具有统计学意义(P<0.01);不同剂量当归多糖组中HT-15/DDP细胞miR-10b-5p表达水平明显低于对照组,TGFBR2表达水平明显高于对照组(P<0.01),随着当归多糖剂量的增加miR-10b-5p表达水平逐渐降低,TGFBR2表达水平逐渐升高(P<0.01)。使用生物信息学数据库(TargetScan、miRanda)预测miR-10b-5p的靶基因,结果显示,TGFBR2是miR-10b-5p的候选靶基因。miR-10b-5p表达下调后,HT-15/DDP细胞中TGFBR2相对表达量明显高于对照组HT-15/DDP细胞中TGFBR2相对表达量(P<0.01)。当归多糖(2.0 mg/mL)+miR-10b-5p inhibitor组、当归多糖(2.0 mg/mL)+TGFBR2 mimics组的HT-15/DDP细胞增殖率、细胞增殖迁移个数明显低于当归多糖组(2.0 mg/mL),HT-15/DDP细胞凋亡率明显高于当归多糖组(2.0 mg/mL)(P<0.05)。结论 当归多糖可通过下调miR-10b-5p表达、上调TGFBR2表达逆转结直肠癌细胞顺铂耐药性,进而抑制肿瘤细胞增殖、迁移,促进细胞凋亡。

当归多糖对低氧环境中骨髓间充质干细胞成骨分化潜能的影响

目的探讨当归多糖对低氧环境中骨髓间充质干细胞(BMSC)成骨分化潜能的影响。方法将BMSC随机分为空白组、低氧组、当归多糖组。使用OriCellTM成人BMSC成骨诱导分化完全培养基对3组细胞进行培养,其中当归多糖组的培养基含有50μg/mL的当归多糖。对空白组BMSC进行常规培养(21%氧浓度),将低氧组与当归多糖组BMSC置于6%氧浓度环境中进行培养。培养14 d后,观察3组的BMSC形态学变化和钙盐沉积情况,检测3组BMSC的成骨分化相关蛋白(骨桥蛋白和骨钙素)表达水平,以及成骨分化关键转录因子RUNT相关转录因子2(RUNX2)表达情况。结果与空白组相比,低氧组BMSC的骨钙素、骨桥蛋白、RUNX2的表达水平均降低(P<0.05),高密度钙结节及钙盐沉积减少。与低氧组相比,当归多糖组BMSC骨钙素、骨桥蛋白、RUNX2的表达升高(P<0.05),高密度钙结节及钙盐沉积增多。结论低氧环境可以抑制BMSC的成骨分化潜能,而当归多糖能够有效维持低氧环境中BMSC成骨分化潜能的稳定。

当归多糖促进骨髓移植重建受体小鼠造血功能

目的探讨当归多糖(ASP)调控骨髓基质细胞(BMSCs)促进供体骨髓移植(BMT)重建受体小鼠造血功能的机制。方法分离纯化8~10周龄雄性C57BL/6J小鼠骨髓单个核细胞(BMMNCs),移植给同龄同种雌性受体小鼠,第9天取受体鼠BMMNCs,再次移植给雌性受体小鼠。移植受体鼠分为对照组:假照射;辐射组:8.0 Gy X射线全身一次性照射;骨髓移植组:照射同辐射组,经尾静脉移植雄性供体BMMNCs(5×10^(6)个细胞);骨髓移植ASP组:同骨髓移植组,从移植的第1天起连续腹腔注射ASP[100 mg/(kg·d)×9]。模型构建期间记录小鼠体重与生存变化,模型构建结束后采集受体小鼠BMMNCs检测Y染色体性别决定区(SRY)基因,测定外周血血常规指标,计算股骨BMMNCs数,观察骨髓组织病理学;分离培养受体鼠BMSCs,观察细胞贴壁生长,5-乙炔基-2’-脱氧尿苷(EdU)法检测BMSCs增殖;分析细胞内活性氧(ROS)水平、超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;检测BMSCs培养上清液中粒细胞-巨噬细胞集落刺激因子(GM-CSF)、干细胞因子(SCF)和胰岛素样生长因子1(IGF-1)水平,检测BMMNCs与各组BMSCs共培养48 h,形成造血祖细胞混合集落(CFU-Mix)数量;Real-time PCR分析受体BMMNCs的Notch信号通路相关基因(Notch1、Jagged1、Hes1)表达。结果单纯辐射组小鼠全部死亡,骨髓移植ASP组受体鼠体重下降不明显;在受体雌鼠BMMNCs中检测到SRY基因;骨髓移植ASP组受体鼠外周血血常规和BMMNCs总数下降幅度不显著,骨髓组织病理学损伤减轻;ASP能促进BMSCs增殖,降低BMSCs中ROS、MDA含量,提高SOD活性;促进BMSCs分泌SCF、GM-CSF及IGF-1,提高BMMNCs与受体BMSCs共培养后的CFU-Mix产率;提高受体小鼠BMMNCs中Notch1、Jagged1、Hes1 mRNA表达。结论ASP促进受体小鼠造血功能重建机制与减轻造血微环境氧化应激损伤,提高BMSCs分泌造血生长因子,调控Notch信号通路有关。

基于NF-κB信号通路探讨当归多糖对LPS介导的SD大鼠骨性关节炎作用研究

目的探究当归多糖(Angelica sinensis polysaccharides,ASP)对骨关节炎软骨细胞的软骨保护和治疗作用。方法通过体外脂多糖(lipopolysaccharide,LPS)诱导来构建关节炎软骨细胞,使用CCK-8法(Cell Counting Kit-8 assay,CCK-8)筛选出实验组当归多糖的剂量,通过活死细胞染色、DNA和糖胺聚糖(glycosaminoglycan,GAG)含量检测、实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测、酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)、免疫荧光染色、番红O固绿及免疫组化染色等来探究当归多糖对关节炎性软骨细胞的保护和治疗效果。结果CCK-8成功筛选出25μg/mL的当归多糖作为实验组。活死细胞染色中实验组细胞活力较模型组好,且实验组DNA含量和GAG分泌量均显著高于模型组(P<0.05)。PCR中实验组的炎症相关基因相对表达量较模型组显著下调(P<0.05),而软骨相关基因相对表达量显著上调(P<0.05)。Elisa显示实验组肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)的分泌水平低于模型组(P<0.05),体内免疫组化染色进一步验证了当归多糖实验组的P65(NF-κB P65)、iNOS及MMP13免疫组化中实验组的阳性表达显著低于模型组。结论当归多糖通过抑制NF-κB信号通路的表达对LPS诱导的骨性关节炎具有一定的治疗作用。

Study on Intervenient Effect of Angelica Sinensis Polysaccharide on Immunological Liver Injury and Its Mechanism in Mice

Objective: To study the changes of expressions of nitric oxide synthase (NOS) of the constitutive type (cNOS) and inducible type (iNOS), the apoptosis related genes bax and bcl 2, as well as the tumor necrosis factor (TNF α) in immunological liver injury (ILI) and to explore the preventive effects of Angelica sinensis polysaccharide (ASP) on ILI and its mechanism in mice.Methods: ILI model mice induced by intraperitoneal injection of lipo polysaccharide (LPS) and BCG vaccine were treated with ASP of different doses (30mg/kg, 60mg/kg) by gastrogavage every day for 7 days. The serum alanine transaminase (ALT) and glutathione S transferase (GST) activities and NO content in the liver were detected; the expressions of cNOS, iNOS, bcl 2, bax were assessed with immuno histochemical method, and the TNF α mRNA expression in the liver was observed by reverse transcriptase polymerase chain reaction (RT PCR).Results: Compared with the normal mice , the NO production and ALT, GST levels were raised significantly in the model mice, the TNF α mRNA expression was also raised significantly. But no obvious changes of cNOS was found. Small dose ASP (30mg/kg) could reduce NO production and ALT, GST levels in model mice by 19.5%, 23.7% and 40.0% respectively, decrease the expression of iNOS and bax by 48.3%, and 26.4%, and increase the expression of cNOS, bcl 2 by 66.9% and 337.3%, respectively, but it could not reduce the TNF α mRNA expression in the liver. Large dose of ASP (60mg/kg) was not more effective than that of small dose.Conclusion: Changes of NO production and TNF α mRNA may play an important role in ILI. The mechanism of ASP in intervening ILI may be through modulation on cNOS, iNOS, bax, bcl 2 expression to block the damage of BCG vaccine and LPS on hepatocytes.

Effect of Angelica Sinensis Polysaccharide-lron Complex on Iron Deficiency Anemia in Rats

Objective： To investigate the therapeutic effects of Angelica sinensis polysaccharideiron complex （APIC） on rats with iron deficiency anemia （IDA）. Methods： The IDA rat model was established by adopting low-iron forage with a small amount of regular bloodletting. The rats were randomly divided into a model group, three AIPC groups （high, middle, and low dosage）, an Angelica sinensis polysaccharide （ASP） group, a mixture group （ASP＋FeCl3） and a positive control group （Niferex）. Changes in hemoglobin （Hb）, red blood cell count （RBC）, hematocrit （HCT） and iron content of whole blood were observed. Results： There was a significant difference before and after administration in all treated groups and all indices were restored to near-normal levels in the APIC groups and the positive control group. There was a significant difference among the changes of the indices in all the APIC groups and those of the model group but not between those of the APIC groups and the positive control group. However, the recovery of the indices in the APIC groups was superior to that in the positive control group. Conclusion： APIC not only has a superior therapeutic effect on IDA, but also has the effect of the ASP on supplementing blood and activating blood circulation. Hence, it may be used as a new iron-supplementing agent with a double therapeutic efficacy on blood supplementation for the treatment of IDA.

Fe_(3)O_(4)@Angelica sinensis polysaccharide nanoparticles as an ultralow-toxicity contrast agent for magnetic resonance imaging

Although iron oxide(Fe_(3)O_(4)) nanoparticles have broad application prospects as magnetic resonance imaging(MRI) contrast agent, their biocompatibility and biotoxicity still need to be improved. In this study, we prepared Fe_(3)O_(4)@Angelica sinensis polysaccharide nanoparticles(Fe_(3)O_(4)@ASP NPs) with a 9 nm Fe_(3)O_(4) core and ASP as the coating material. The Fe_(3)O_(4)@ASP NPs are superparamagnetic, can be taken up by liver and spleen macrophages in the circulatory system in vivo, and are a good-biocompatibility and low-toxicity transverse relaxation time(T_(2)) and T_(2)-star(T_(2)^(*)) magnetic resonance imaging(MRI) contrast agent for the liver. The cytotoxicity assessment using HeLa cells and the pathological tests in mice validate that Fe_(3)O_(4)@ASP NPs have low toxicity and good biocompatibility in vivo, which can be attributed to the ASP as a natural polysaccharide with good biocompatibility and its function of protecting the liver. Fe_(3)O_(4)@ASP NPs are a potential new MRI contrast agent with high signal intensity in vivo.

当归多糖调控肠道菌群对脑缺血再灌注小鼠的影响

目的 探讨当归多糖(Angelica Sinensis Polysaccharide,ASP)改善小鼠肠道微生态对脑缺血再灌注(Ischemia/reperfusion injury,I/R injury)的影响。方法 将雄性SPF级昆明小鼠随机分为假手术组(Sham组),大脑中动脉栓塞组(MCAO组),当归多糖灌胃组(ASP组),粪菌移植组(Fecal microbiota transplantation,ASP-FMT),每组16只。连续给药7 d后建立MCAO模型,24 h后使用mNSS法检测小鼠神经功能缺陷评分;TTC染色法检测脑梗死相对面积;试剂盒法测定右侧大脑缺血半暗带组织SOD、MDA、GSH含量;Western blot测定炎症因子IL-1β、IL-5、IL-6、IL-10表达量;盲肠内容物16s rRNA测序分析肠道微生物多样性。结果 与MCAO组相比,ASP干预后小鼠神经功能明显改善(P<0.001),脑梗死相对面积明显减少(P<0.01),氧化应激水平下降(P<0.05);与MCAO组相比,ASP降低了脑组织IL-1β、IL-5及IL-6表达、上调IL-10表达量(P<0.05);MACO后小鼠肠道菌群丰富度明显减少,益生菌比例下调,当归多糖灌胃或菌群移植均可部分恢复菌群丰富度,上调益生菌比例;肠道菌群的变化与脑组织损伤程度具有相关性。结论 当归多糖通过改善肠道微生态,降低氧化应激水平,下调炎症因子水平,减轻脑缺血再灌注后神经功能损伤,减少梗死范围。

超声辅助低共熔溶剂提取当归药渣粗多糖的工艺优化

为研究当归(Angelica sinensis)药渣提取粗多糖的工艺,本研究以当归药渣粗多糖(Angelica sinensis Residue Crude Polysaccharide, ARP)的提取率为指标,考察低共熔溶剂(Deep Eutectic Solvent, DES)的种类、组成成分的摩尔比、DES含水量、提取温度、提取时间和液料比对ARP提取率的影响,并结合响应面分析法优化提取工艺参数。研究结果表明,氯化胆碱和乙二醇组成的低共熔溶剂更适合提取ARP,最优提取工艺为提取温度82℃、提取时间40 min、摩尔比6∶1。该工艺条件下的ARP提取率为(14.43±0.10)%,ARP总糖含量为(47.54±1.23)%;而传统水提醇沉法的ARP提取率为(9.85±0.12)%,ARP总糖含量为(25.45±1.37)%。因此,氯化胆碱和乙二醇组成的低共熔溶剂是一种较好的提取溶剂,可替代传统溶剂进行多糖的提取,具有实际应用价值。

当归多糖对小鼠脾淋巴细胞增殖及诱生IFN-γ的影响

目的 研究当归多糖对小鼠脾淋巴细胞增殖及诱生IFN γ的影响。方法 用 4种不同浓度乙醇将当归总多糖 (AP)进行分级沉淀 ,获得AP I ,AP II,AP III和AP IV 4个部分多糖。用 [3 H] TdR参入法检测当归部分多糖对小鼠脾淋巴细胞增殖作用 ;结晶紫染色法测定IFN γ的生物活性 ;ELISA法测定IFN γ的含量。结果 AP I和AP II能明显促进体内外小鼠脾淋巴细胞增殖 ,体外显著诱导脾淋巴细胞分泌IFN γ ,增强IFN γ的生物活性 ;特别是AP II体外诱生较高含量的IFN γ ,AP I体内诱生的IFN γ生物活性较强。结论 AP I和AP II是免疫活性较强的部分。

当归多糖的分离、纯化及分析鉴定

目的 从新鲜中国当归 [Angelica sinensis(Oliv)Diels]中分离纯化出多个多糖组分 ,并对其理化性质进行分析鉴定 .方法 采用水煮—乙醇沉淀法从新鲜当归中提取当归总多糖 AP- 0 ;AP- 0用 80 g· kg- 1十六烷基三甲基溴化铵(CTAB)沉淀得到当归多糖亚组分 AP- 1;CTAB处理后的上清用 10 g· kg- 1 H3BO3,2 mol· L- 1 Na OH处理得到当归多糖亚组分 AP- 2 ;所剩上清 ,用乙酸处理后 ,再用乙醇沉淀 ,得到当归多糖亚组分 AP- 3.总糖含量测定采用改良苯酚 -硫酸法 ;蛋白质含量测定采用 Serva Blue- G染料结合法 ;糖醛酸含量测定采用间羟基连苯法 .采用新华中速滤纸纸色谱和硅胶G薄层色谱分析多糖各组分的单糖组成 .高效凝胶过滤色谱和高效离子交换色谱分析多糖各组分的 Mr分布和电荷性质 .红外光谱法分析多糖各组分的糖苷键构成 .结果 AP- 0总糖 970 g· kg- 1 ,其中糖醛酸 2 10 g· kg- 1 ,蛋白质 30g·kg- 1 ;AP- 1总糖 970 g· kg- 1 ,其中糖醛酸 172 g· kg- 1 ,蛋白质 30 g· kg- 1 ;AP- 2总糖 830 g· kg- 1 ,其中糖醛酸 2 5 7g·kg- 1 ,蛋白质 170 g· kg- 1 ;AP- 3总糖 970 g· kg- 1 ,其中糖醛酸 86 g· kg- 1 ,蛋白质 30 g· kg- 1 .AP- 0 ,AP- 1,AP- 2和 AP- 3主要由葡萄糖 。