黄芪甲苷对体外培养神经干细胞分化作用的研究

目的观察黄芪甲苷对体外培养神经干细胞分化作用的影响。方法利用无血清培养技术在体外对新生Wistar大鼠海马区与神经干细胞进行培养,观察相同接种密度条件下,不同浓度黄芪甲苷（20mg／L,40ms／L,80mg／L）对神经干细胞分化作用的影响,采用免疫细胞化学法鉴定神经干细胞及其分化后细胞。结果从新生大鼠海马区分离出的神经干细胞经鉴定呈nestin阳性,且加入黄芪甲苷组所表达的神经干细胞和胶质纤维酸性蛋白阳性细胞较对照组明显增多（17．60±2．85vs15．65±2．29;24．10±3．13VS21．95±2．37;P〈0．05）。结论黄芪甲苷对体外培养大鼠海马区神经干细胞的分化有促进作用。     

黄芪甲苷抗凋亡作用机制的研究进展

黄芪甲苷是黄芪的主要活性成分,具有众多的生物活性,如抗神经退行性疾病（阿尔海默病,帕金森）、抗应激、抗纤维化、抗病毒、抗炎等。通过查阅国内外文献,发现抗细胞凋亡作用与多种疾病密切相关。本文从细胞凋亡产生的重要途径如死亡受体途径、线粒体途径、内质网途径、PI3K/Akt途径、MAPK途径等多个方面对黄芪甲苷抗凋亡作用机制进行综述,为黄芪甲苷在抗细胞凋亡的相关研究提供参考。     

黄芪甲苷对体外神经干细胞增殖作用影响的研究

目的探讨黄芪甲苷(astragaloside,AS)对神经干细胞增殖的影响。方法体外培养大鼠胎鼠神经干细胞,Nestin免疫化学染色鉴定NSCs,BrdU标记鉴定NSCs增殖。BrdU标记免疫细胞化学染色检测神经干细胞增殖能力。实时定量PCR技术探讨黄芪甲苷促进NSCs增殖的作用机制。结果Nestin鉴定为阳性,Brdu标记亦呈阳性;BrdU标记结果表明,黄芪甲苷高、中、低剂量组NSCs的增殖率明显提高(P<0.05,P<0.01)。实时定量PCR结果表明在诱导NSCs增殖过程中,黄芪甲苷高剂量组Hes5基因表达增加(P<0.05)。结论黄芪甲苷对NSCs增殖具有明显的诱导作用,其可能通过上调与细胞增殖相关基因Hes1、Hes5、cyclinD1表达而起作用,但也可能调节与其它增殖通路有关。     

黄芪甲苷对心脏保护作用的研究现状

黄芪甲苷通过抑制核因子-кB(NF-кB)通路、降低NF-кB的表达水平来抑制缺氧心肌细胞的凋亡。黄芪甲苷通过调节自噬基因、上调B细胞淋巴瘤/白血病-2基因(Bcl-2),降低Bcl-2相关X蛋白(Bax)及半胱氨酸蛋白酶(caspase)-3的表达以及上调低氧诱导因子-1α(HIF-1α)表达等途径对缺血再灌注心脏发挥保护作用。黄芪甲苷也可降低心肌转化生长因子β1(TGF-β1)、Smad2/3和Smad4蛋白表达,抑制心肌纤维化,保护心脏。黄芪甲苷对异丙肾上腺素致心脏损伤、对血管紧张素Ⅱ致心脏损伤、对高血糖性心脏损伤都具有一定的保护作用。     

黄芪甲苷对肾脏保护作用机制的研究进展

黄芪甲苷是中药黄芪的主要活性成分之一,研究证实具有抗氧化应激、抗炎、抗纤维化等作用,能保护肾脏并延缓慢性肾病进展,有望开发成治疗肾脏疾病的新药。本文将对近年黄芪甲苷保护肾脏的作用机制作一综述。     

黄芪甲苷体外抗腺病毒作用研究

目的研究黄芪甲苷体外抗腺病毒作用。方法采用细胞病变效应(CPE)抑制实验和噻唑蓝(MTT)比色法观察黄芪甲苷对人腺病毒3型(HAdV3)的抑制作用,激光共聚焦显微镜荧光分析检测黄芪甲苷在病毒生物合成阶段对HAdV3六邻体(hexon)蛋白表达的影响。结果 CPE及MTT结果表明黄芪甲苷对腺病毒有直接灭活作用,同时能够抑制腺病毒的复制和吸附,病毒抑制率与药物浓度呈正相关,但在细胞保护作用方式下黄芪甲苷不能阻断病毒进入细胞。在生物合成阶段黄芪甲苷组与病毒对照组比较hexon蛋白的表达明显降低。结论黄芪甲苷在体外具有抗腺病毒作用,黄芪甲苷抗腺病毒作用可能与其在生物合成阶段抑制hexon蛋白的表达有关。     

黄芪甲苷对大鼠海马源神经干细胞体外增殖的影响

目的观察黄芪甲苷对神经干细胞（NSC）体外增殖的影响,为神经干细胞应用于临床提供理论依据。方法利用无血清培养技术,从新生大鼠海马区分离培养NSC,并进行体外扩增、传代。取传至第3代的NSC,观察不同浓度黄芪甲苷（40、80、120mg／L）对其增殖的影响。对神经球形成数进行计数并进行噻唑蓝（MTT）比色分析。结果40mg／L和80mg／L黄芪甲苷实验组神经球形成数和细胞吸光度值均明显高于对照组（神经球形成数：8．1±1．4,8．8±1．3比7．2±1．2;细胞吸光度值：0．87±0．02、0．89±0．03比0．694-0．03,P〈0．05或P〈0．01）,而120mg／L黄芪甲苷组实验组神经球形成数和细胞吸光度值（分别为3．1±1．2、0．30±0．02）低于对照组（P〈0．01）。结论在一定浓度范围内,黄芪甲苷可促进体外培养NSC的增殖。     

黄芪甲苷IV对TGF-β1/Smad2介导的癫痫合并抑郁小鼠的保护作用

目的 探讨黄芪甲苷IV对癫痫合并抑郁小鼠的治疗和保护作用及机制.方法 60只C57/BL6小鼠,随机分为对照组、癫痫组、黄芪甲苷IV低剂量组(20 mg/kg)、黄芪甲苷IV高剂量组(40 mg/kg)、黄芪甲苷IV+LY2109761组(40 mg/kg+1μg/只),每组12只.以氯化锂(180 mg/kg)联合匹...     

黄芪多糖与黄芪甲苷配伍对辐射损伤模型小鼠的保护作用

目的:研究黄芪多糖与黄芪甲苷配伍对辐射损伤模型小鼠的保护作用。方法:100只雌性ICR小鼠随机分为正常对照(等容生理盐水)组、黄芪多糖对照(80 mg/kg)组、黄芪甲苷对照(80 mg/kg)组、模型(等容生理盐水)组、黄芪多糖(80 mg/kg)组、黄芪甲苷(80 mg/kg)组与联合用药①、②、③、④(黄芪多糖80、40、20、80 mg/kg+黄芪甲苷20、40、80、80 mg/kg)组。灌胃给药,每天1次,连续14 d。末次给药后小鼠接受60Coγ射线(剂量:5 cy)一次性全身辐射以复制辐射损伤模型。测定小鼠30 d存活率和死亡小鼠平均存活时间;肝脏HE染色光镜下观察小鼠肝组织形态学;测定小鼠外周血白细胞数、骨髓细胞DNA含量、血清超氧化物歧化酶(SOD)活性。结果:与正常对照组比较,模型组小鼠30 d存活率降低,死亡小鼠平均存活时间缩短;小鼠肝细胞广泛肿胀,出现大滴性脂肪变、气球样变,肝脏细胞点状坏死严重,炎性因子大面积浸润组织;外周白细胞计数减少;骨髓细胞DNA含量减少;血清SOD活性减弱,差异具有统计学意义(P<0.01)。与模型组比较,联合用药①、②、③、④组小鼠30 d存活率升高,死亡小鼠平均存活时间延长,外周白细胞计数增加,骨髓细胞DNA含量增加,小鼠肝细胞形态学明显改善;联合用药①、②、③组小鼠血清中SOD活性增强,差异具有统计学意义(P<0.01)。结论:黄芪多糖与黄芪甲苷配伍是通过多靶点发挥作用,其中黄芪多糖与黄芪甲苷质量比为4∶1时配伍效果最佳。     

Effects of astragaloside IV on the pharmacokinetics of puerarin in rats

Abstract

1. Radix astragali and puerarin are always used together for cardiovascular disease in China clinics.

2. This study investigates the effects of astragaloside IV (AS-IV, the main components of radix astragali) on the pharmacokinetics of puerarin in rats.

3. The pharmacokinetics of orally administered puerarin (50?mg/kg) with or without AS-IV pretreatment (100?mg/kg/day for seven days) were investigated. The plasma concentration of puerarin was determined using LC–MS/MS method, and the pharmacokinetics profiles were calculated and compared. Caco-2 cell transwell model was also used to investigate the effects of AS-IV on the transport pf puerarin.

4. The results showed that when the rats were pretreated with AS-IV, the maximum concentration (C_max) of puerarin decreased from 760 to 467?ng/mL (p?<?.05, n?=?6, 90% CI, 293?±?61.28), and the area under the concentration-time curve from zero to infinity (AUC_0–inf) also decreased from 4097 to 2330?μg·h/L (p?<?.05, n?=?6). The oral clearance of puerarin increased significantly from 11.9 to 22.4?L/h/kg (p?<?.05, n?=?6). The Caco-2 cell transwell experiments indicated that AS-IV could increase the efflux ratio of puerarin from 1.81 to 2.79 through inducing the activity of P-gp.

5. In conclusion, these results indicated that AS-IV could affect the pharmacokinetics of puerarin, possibly by decreasing the systemic exposure of puerarin by inducing the activity of P-gp.

     

黄芪皂苷Ⅳ对小鼠T、B淋巴细胞增殖和腹腔巨噬细胞功能的影响

目的：探讨黄芪皂苷Ⅳ（ASI）对小鼠T,B淋巴细胞增殖和腹腔巨噬细胞分泌的细胞因子的影响。方法：采用MTT法检测T,B淋巴细胞的增殖;采用分光光度计测定法检测抗体活性;IL－1活性用胸腺细胞增殖法测定;TNF－α活性用L9292细胞杀伤法测定。结果：1）ASI50－200mg/kg ig7天能够促进T淋巴细胞增殖和抗体生成,而ASI50－100mg/kg 能够促进B淋巴细胞增殖,但是200mg/kg对B淋巴细胞增殖无影响;（2）ASI体外仅在200nmol/L对T,B淋巴细胞有促进作用;（3）ASI 1nmol/L可以促进腹腔巨噬细胞分泌IL－1,而100－1000nmol/L则抑制腹腔巨噬细胞分泌IL－1;（4）ASI体外可以抑制LPS刺激或无LPS刺激下的腹腔巨噬细胞分泌TNF－α。结论：ASI能够促进小鼠T,B淋巴细胞的增殖和抗体生成,同时可以抑制腹腔巨噬细胞体内分泌IL－1和TNF－α。     

黄芪甲苷对黑色素瘤细胞生长和迁移的影响

目的研究黄芪甲苷对黑色素瘤A375细胞生长和迁移的作用。方法通过MTT试验检测不同浓度的黄芪甲苷对A375细胞增殖的影响,应用流式细胞术对黄芪甲苷作用后的A375细胞周期进行分析。通过Annexin V-FITC/PI双染色法观察经黄芪甲苷处理的A375细胞的凋亡情况,transwell实验分析黄芪甲苷对A375细胞迁移的作用。通过Western Blot验证黄芪甲苷对CDK4/CDK6,caspase 3和MMP2的作用。结果不同浓度的黄芪甲苷对A375细胞的增殖均有一定的抑制作用。黄芪甲苷作用后细胞周期阻滞于G1期,细胞凋亡增加,迁移能力减弱。黄芪甲苷通过抑制细胞CDK4/CDK6,caspase 3和MMP2的表达来抑制细胞增殖和迁移,促进细胞凋亡。结论黄芪甲苷能够抑制A375细胞的增殖,促进其凋亡并且抑制其迁移作用,可考虑用于黑色素瘤的预防和治疗。     

Effects of the phosphodiesterase IV inhibitor rolipram on Th1 and Th2 immune responses in mice

The present study was designed to investigate the effect of the phosphodiesterase IV inhibitor rolipram on Th1 and Th2 immune responses in mice. Mice were immunized subcutaneously at the base of the tail with ovalbumin (OVA) emulsified with complete Freund's adjuvant (day 0) and were treated daily with oral administration of various doses of rolipram from days 0 to 20. On day 21, production of anti-OVA IgG and proliferative responses to the antigen were determined. Anti-OVA IgG2a and interferon-gamma (IFN-gamma), as indicators of Th1 responses, and anti-OVA IgG1 and interleukin-10 (IL-10), as indicators of Th2 responses, were also measured. The results showed that treatment with rolipram failed to affect the production of OVA-specific IgG but decreased the proliferation of spleen cells to the antigen. Its inhibitory effect on these immune responses was correlated with a marked decrease in IFN-gamma but not IL-10 production, although neither anti-OVA IgG2a nor IgG1 production was affected by rolipram. These results suggest that rolipram may preferentially inhibit Th1 responses more effectively than Th2 responses. Administration of rolipram resulted in suppression of antigen (OVA)-induced arthritis in mice. The suppression of joint inflammation by rolipram was associated with the inhibition of the OVA-specific proliferative responses of spleen cells and IFN-gamma secretion. These results indicate that rolipram may be effective in regulating Th1-mediated diseases such as rheumatoid arthritis.     

Absorption enhancement study of astragaloside IV based on its transport mechanism in caco-2 cells.

The purpose of this study was to investigate the transport characteristics and mechanisms for discovering the possible causes of the low bioavailability of astragaloside IV and to develop an absorption enhancement strategy. Caco-2 cells used as the in vitro model. Results showed a low permeability coefficient (3.7 x 10(-8)cm/s for transport from the AP to BL direction), which remained unchanged throughout the concentration range studied, indicating that the transport of astragaloside IV was predominantly via a passive route. The AP to BL transport of astragaloside IV was found to be highly sensitive to the extracellular Ca2+ concentration, which suggested that its transport may be via a paracellular route. Both chitosan and sodium deoxycholate can increase the permeation efficiency of astragaloside IV. This study indicated that astragaloside IV having a low fraction dose absorbed in humans mainly due to its poor intestinal permeability, high molecular weight, low lipophilicity as well as its paracelluar transport may directly result in the low permeability through its passive transport. Meanwhile, chitosan and sodium deoxycholate can be used as absorption enhancers based on its transport mechanism.     

黄芪皂苷IV对缺血大鼠心肌钙运转和心功能的影响

目的：研究黄芪黄苷IV（XGA）对缺血大鼠心肌钙运转和心功能的影响。方法：84只Wistar大鼠分为3组：对照组（12只）;缺血组（12只）,皮下注射异丙肾上腺素（30mg/kg);XGA组（60只）,皮下注射异丙肾上腺素后给予XGA,测定大鼠血流动力学指标、心肌功能和心肌细胞内外钙的变化,观察XGA对缺血大鼠心肌钙运转和心功能的量－效和时－效关系。结果：给予XGA后缺血大鼠心功能和血流动力学明显改善,随着XGA剂量的增加和XGA作用时间的延长,缺血大鼠的心输出量、心率、每博量、平均动脉压、动脉收缩压、左心室博出功、左室和右室收缩末期压力和右心室的 dp/dt逐渐恢复到对照组水平;给予XGA后心肌细胞游离钙和心肌组织总钙与缺血组比较明显下降,而红细胞膜钙泵活性较钙血组明显增加,但心肌细胞游离钙和心肌组织总钙下降的幅度及红细胞膜钙泵活性增加的幅度并没有XGA剂量的增加而增加;随XGA作用时间的延长,心肌细胞游离钙逐渐下降。结论：XGA能够明显改善缺血大鼠的心功能,减少心肌细胞内过多的钙积聚是其作用机制之一。     

黄芪皂苷Ⅳ对缺血大鼠心肌钙运转和心功能的影响(英文)

目的:研究黄芪皂苷IV(XGA)对缺血大鼠心肌钙运转和心功能的影响.方法:84只Wistar大鼠分为3组:对照组(12只);缺血组(12只),皮下注射异丙肾上腺素(30 mg/kg);XGA组(60只),皮下注射异丙肾上腺素后给予XGA.测定大鼠血流动力学指标、心脏功能和心肌细胞内外钙的变化,观察XGA对缺血大鼠心肌钙运转和心功能的量-效和时-效关系.结果:给予XGA后缺血大鼠心功能和血流动力学明显改善,随着XGA剂量的增加和XGA作用时间的延长,缺血大鼠的心输出量、心率、每博量、平均动脉压、动脉收缩压、左心室搏出功、左室和右室收缩末期压力和右心室的 dp/dt逐渐恢复到对照组水平;给予XGA后心肌细胞游离钙和心肌组织总钙与缺血组比较明显下降,而红细胞膜钙泵活性较缺血组明显增加,但心肌细胞游离钙和心肌组织总钙下降的幅度及红细胞膜钙泵活性增加的幅度并没有随XGA剂量的增加而增加;随XGA作用时间的延长,心肌细胞游离钙逐渐下降.结论:XGA能够明显改善缺血大鼠的心功能,减少心肌细胞内过多的钙积聚是其作用机制之一.     

心安胶囊中黄芪甲苷含量测定方法的研究

目的测定心安胶囊中黄芪甲苷的含量,建立该制剂中黄芪甲苷的高效液相色谱法测定方法。方法采用高效液相色谱法,用十八烷基硅烷键合硅胶为填充剂;以乙腈-水(35∶65,V/V)为流动相;蒸发光散射检测器检测;理论塔板数,按黄芪甲苷峰计应不低于4000。结果黄芪甲苷进样量在2.44～14.64μg范围内与峰面积呈良好的线性关系。Y=1.323X+13.233,r=1.000。平均加样回收率100.2%,RSD为3.49%。结论该方法结果准确、重现性好,可用于测定心安胶囊中黄芪甲苷的含量。