Anti-inflammatory and DNA Repair Effects of Astragaloside IV on PC12 Cells Damaged by Lipopolysaccharide

Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside IV on key targets using high-throughput sequence technology and bioinformatics analyses.Methods PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25,0.5,0.75,1,and 1.25 mg/mL for 24 h.Cell morphology was evaluated,and cell survival rates were calculated.A neurocyte inflammatory model was established with LPS treatment,which reached a 50%cell survival rate.PC12 cells were treated with 0.01,0.1,1,10,or 100µmol/L astragaloside IV for 24 h.The concentration of astragaloside IV that did not affect the cell survival rate was selected as the treatment group for subsequent experiments.NOS activity was detected by colorimetry;the expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting.The differentially expressed genes(DEGs)between the groups were screened using a second-generation sequence(fold change>2,P<0.05)with the following KEGG enrichment analysis,RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells.Results The viability of PC12 cells was not altered by treatment with 0.01,0.1,or 1µmol/L astragaloside IV for 24 h(P>0.05).However,after treatment with 0.5,0.75,1,or 1.25 mg/mL LPS for 24 h,the viability steadily decreased(P<0.01).The mRNA and protein expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS,and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h(P<0.01);however,these changes were reversed when PC12 cells were pretreated with 0.01,0.1,or 1µmol/L astragaloside IV in PC12 cells and then treated with 1 mg/mL LPS for 24 h(P<0.05).Second-generation sequencing revealed that 1026 genes were upregulated,while 1287 genes were downregulated.The DEGs were associated with autophagy,TNF-α,interleukin-17,MAPK,P53,Toll-like receptor,and NOD-like receptor signaling pathways.Furthermore,PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2,CCL11,CCL7,MMP3,and MMP10,which are associated with the IL-17 pathway.RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results.Conclusion LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage.astragaloside IV plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.

Astragaloside IV Ameliorates Inflammatory Damage in Mice with Acute Liver Failure

Acute liver failure is a life-threatening clinical syndrome with a high mortality rate. Currently, the research on Astragaloside IV in liver diseases primarily focuses on liver cancer, and there is limited understanding of its mechanism in acute liver failure’s innate immunity. Therefore, this study aims to investigate the potential protective effect of Astragaloside IV on acute liver failure and its impact on innate immune cells. The study employed D-GalN/LPS-induced acute liver failure mouse models and employed various techniques such as a range of molecular and analytical techniques. The experimental results demonstrated that treatment with Astragaloside IV significantly reduced the inflammatory response, alleviated liver injury, and improved the survival rate of mice with acute liver failure induced by D-GalN/LPS. Further investigations revealed that AS-IV played a beneficial role by regulating the proportion of CD11b+Ly6Chi monocytes and the secretion of inflammatory cytokines and anti-inflammatory metabolites. These findings suggest that the pharmacological mechanism of AS-IV may involve targeted regulation of CD11b+Ly6Chi monocytes in both peripheral blood and liver. The implications of this study’s results are twofold. Firstly, they provide a basis for the clinical application of AS-IV in treating liver failure, offering potential therapeutic benefits. Secondly, they serve as a reference for further development of safer and more effective modified compounds.

Determination of Astragaloside IV in Yupingfeng Oral Solution by HPLC-ELSD Method

[Objective] This study aimed to establish a method for determining the content of Astragaloside IV in Yupingfeng oral solution.[Method] The HPLC-ELSD method was adopted.The chromatographic column was Venusil MP（4.6 mm × 150 mm,5 μm）.The mobile phase was acetonitrile-water（35∶65）.The ELSD evaporator tube temperature was 65 ℃.N2 was used as the carrier gas（pressure,30 psi）.[Result] When the content of Astragaloside IV ranged from 0.5 to 5.0 μg,the Astragaloside IV content showed a good linear relationship with peak area（r=0.999,n=6）.The average recovery was 96.36%,and the RSD was 2.46%.[Conclusion] This method is accurate and reliable,and can be applied in the quality control of Yupingfeng oral solution.

Astragaloside IV prevents high-fat diet induced obesity partially through enhancing leptin signaling transduction

Aim To investigate the effects of astragaloside IV （ASI） on high-fat diet （HFD） induced obese mice. Methods The male mice aged 6 weeks were randomly divided into three groups （u- 18/group）, namely control group, model group and ASI-treated group. Control group were fed with standard diet, whereas the other two groups were given high fat diet. ASI-treated mice were daily intraperitoneally injected with ASI （25 nag · kg^-1）. Mean- while, the other group mice were treated with saline. Body weight of mice was monitored every week and lasted for 13 weeks. Serum cholesterol and triglyceride content were measured with respective kits. Serum leptin level was deter- mined by ELISA kit. Expression of leptin receptor in hypothalamus was measured by Western blot assay. Gene ex- pression of neuropeptide Y （NPY） and agouti-related protein （AGRP） in hypothalamus was detected by qPCR assay. In addition, leptin receptor-deficient db/db mice were given intraperitoneally with ASI （25 mg ~ kg-1） or saline for 13 weeks （u- 8/group）. Results ASI blocked body weight gain, suppressed appetite, improved leptin resistance, lowered serum triacylglycerol （TG） and total cholesterol （TC） contents, reduced accumulation of fat tissues and pre- vented enlargement of adipose cells in HFD fed mice. Furthermore, ASI increased the protein expression level of lep- tin receptor in hypothalamus, and inhibited the mRNA expression levels of NPY and AGRP. However, ASI could not decrease body gain in leptin receptor - deficient db/db mice as well as the mRNA expression levels of NPY and AGRP. Conclusion The study suggested that ASI could efficiently prevent HFD-induced obesity in C57BL/6 mice,which was partially mediated through enhancing leptin signaling transduction.

Protective effect of astragaloside IV on cardiac hypertrophy in rats and its mechanism

Objective:To investigate the effect of astragaloside IV on cardiac hypertrophy and its regulation on autophagy.Methods:Fifty male Sprague-Dawley rats were randomly divided into sham operation group and abdominal aortic coarctation group(AAC group).There were 10 rats in sham operation group and 40 rats in the AAC group.One week after the operation,there were 32 rats in AAC group,10 rats in sham group.AAC group was randomly divided into model group,low-dose astragaloside group,high-dose astragaloside group and rapamycin group,8 rats in each group.Rapamycin group was a positive autophagy contrast agent group.They were given the corresponding solvents once a day by gavage for six weeks.At the end of study,three rats were randomly selected from each group,left ventricular mass index(LVW/BW),cardiac mass index(HW/BW)and the content of hydroxyproline were measured.HE staining,masson staining and sirius red staining were used to observe the morphological changes of myocardium.The expression of LC3II,LC3I,Beclin1,AMPK and mTOR were detected by western blot.Results:Compared with the sham operation group,AAC group showed hypertrophy,LVW/BW,HW/BW,HYP and p-mTOR/mTOR were significantly increased(P<0.05),p-AMPK/AMPK,LC3II/LC3I,Beclin1 were significantly decreased(P<0.05).Compared with the model group,the low-dose astragaloside IV group showed the hypertrophy of cardiomyocytes was relatively light,LVW/BW and HW/BW were significantly decreased(P<0.05),there was no significant difference in HYP and p-mTOR/mTOR(P>0.05),LC3II/LC3I,Beclin1 and p-AMPK/AMPK were significantly increased(P<0.05).Compared with the model group,high-dose astragaloside IV group and rapamycin group showed reduced myocardial hypertrophy,LVW/BW,HW/BW,HYP and p-mTOR/mTOR were significantly decreased(P<0.05),LC3II/LC3I,Beclin1 and p-AMPK/AMPK were significantly increased(P<0.05).Compared with the low-dose astragaloside group,the high-dose astragaloside group showed reduced myocardial hypertrophy,there were significant differences in each index(P<0.05).Compared with rapamycin group,there was no obvious difference in morphology and structure of myocardial cells,LVW/BW,HYP and p-mTOR/mTOR were decreased(P<0.05),HW/BW and p-AMPK/AMPK had no significant difference(P>0.05),LC3II/LC3I and Beclin1 were increased in high-dose astragaloside group(P<0.05).Conclusion:As IV has protective effect on cardiac hypertrophy in a dose-dependent manner and its mechanism may be related to regulate autophagy.

Determination of the Content of Astragaloside IV in Yikangshu Granules by High Performance Liquid Chromatography

[Objectives]The research aimed to establish a high performance liquid chromatography method for the content determination of astragaloside IV in Yikangshu Granules.[Methods]Kromasil 5μm C18(2),100 A,250 mm×4.6 mm was used;the mobile phase was acetonitrile-water(32∶68);flow velocity was 1.0 mL/min;the temperature of evaporator and sprayer was 80 and 30℃;the column temperature was set at 30℃,and injection volume was 20μL.[Results]Astragaloside IV showed a good linear relationship in the range of 1.01-10.14μg with the peak area,and regression equation was lgY=1.7728lgX+1.597(r=0.9999).The limit of detection for astragaloside IV was 1.96 ng,and the average recovery rate was 95.31%.[Conclusions]This method is sensitive,accurate and reproducible,with good linearity,and it is suitable for the content determination of astragaloside IV in health food Yikangshu Granules.

Potential Role of Astragaloside IV in the Treatment of Fungal Keratitis

Fungal keratitis (FK) is a worldwide visual impairment disease. The pathogenesis of fungal keratitis involves fungi, corneal cells, inflammatory cell infiltration, collagen degradation, inflammatory cytokines and their interactions. Accumulated evidence indicated that Astragaloside IV (AS-IV) possesses a broad range of pharmacological properties, such as efficacy in anti-inflammation, alleviating fibrosis, and immunomodulatory effects. This paper summarizes new findings regarding AS-IV in immune and inflammatory diseases and analyzes the perspective application of Astragaloside IV in fungal keratitis.

Astragaloside IV Alleviates Acute Liver Failure Induced by D-GalN/LPS by Upregulating Autophagy and Reducing Inflammation

Acute liver failure(ALF)is a life-threatening condition that manifests in an extremely serious manner and progresses rapidly.The following study investigated the protective effect of astragaloside IV(AS-IV),a traditional Chinese drug,on ALF,and its underlying mechanisms,focusing on autophagy and inflammation regulation.Mice were randomly divided into a saline group,a D-galactosamine and lipopolysac-charide(D-GalN/LPS)group and an AS-IV group.Biochemical analysis,immunohistochemistry,cytometric bead array,high-throughput quantitative PCR,flow cytometry and Western analysis were used to assess inflammation and liver damage 5 hours after D-GalN/LPS ex-posure.Astragaloside IV treatment reduced mortality by alleviating D-GalN/LPS–induced hepatic damage and decreasing inflammation(decreasing Ly6c+monocyte levels,reducing inflammatory cytokines and increasing anti-inflammatory factors)as well as upregulating au-tophagy.Furthermore,PCR array was used to detect expression of autophagy-related genes,which demonstrated a Log2 fold change in gene expression between the AS-IV and D-GalN/LPS groups ranging from 1.19 to-3.53,with Tnfsf10 showing the largest alteration be-tween the two groups.These data suggest that AS-IV may alleviate ALF by upregulating autophagy and reducing inflammation,and it may therefore be an interesting drug for alleviating ALF.

Content Determination of Astragaloside Ⅳ in Astragalus from Three Different Regions by HPLC

[Objectives] The content of astragaloside in Astragalus membranaceus(Fisch.) Bge.var.mongholicus(Bge.) Hisao from three different regions was determined.[Methods] Referring to the method recorded in the Chinese Pharmacopoeia(2015 edition),the content of astragaloside IV in A.membranaceus was determined by HPLC.[Results] There were great differences in the astragaloside IV content of A.membranaceus among different regions.The content of astragaloside IV in A.membranaceus cultivated in Inner Mongolia was highest(0.155%),followed by that(0.143%) in A.membranaceus cultivated in Gansu,and the content of astragaloside IV in A.membranaceus cultivated in Shanxi was lowest(0.080%).The contents of astragaloside IV in A.membranaceus from different regions were all in line with the standard(not less than 0.040%) of Chinese Pharmacopoeia(2015 edition).[Conclusions]The content of astragaloside IV in A.membranaceus cultivated in three different regions met the medicinal standards.

黄芪活性成分生理功能及在食品中的应用研究进展

黄芪作为最为常用的中药材,含有多种活性成分,其中最主要的有黄芪甲苷、黄芪黄酮和黄芪多糖。这些成分赋予黄芪提高机体免疫力、抗疲劳、消炎、保护心脑血管、保护内脏组织、抗癌抗肿瘤等多种药理作用,文章从黄芪活性成分的生理功能及在食品中的应用研究进展等方面进行综述,以期为黄芪及其活性成分在食品工业中的进一步开发应用提供理论参考。

黄芪甲苷通过激活JNK/p38 MAPK信号通路对急性脑梗死模型大鼠神经功能的影响

目的分析黄芪甲苷通过激活c-Jun氨基末端激酶(JNK)/p38丝裂原活化蛋白激酶(p38 MAPK)信号通路对急性脑梗死模型大鼠神经功能的影响。方法选取50只健康SPF级SD大鼠,10只作为对照组,其余40只建立急性脑梗死模型,其中30只大鼠建模成功,将30只大鼠随机分为模型组、药物对照组、黄芪甲苷组,每组10只对建模成功的大鼠进行给药处理,对照组、模型组大鼠采用0.9%氯化钠溶液灌胃,药物对照组给予20 mg/kg阿托伐他汀灌服,黄芪甲苷组给予70 mg/kg黄芪甲苷应用液灌服,均灌胃12周。观察4组大鼠病理组织学、神经功能[神经元特异性烯醇化酶(NSE)、星形胶质源性蛋白(S100β)]、氧化应激指标[超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)、活性氧(ROS)]、炎性因子[肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)、C反应蛋白(CRP)]、JNK/p38 MAPK通路相关mRNA表达量、JNK/p38 MAPK通路。结果与对照组比较,模型组、药物对照组、黄芪甲苷组SOD、CAT降低,NSE、S100β、MDA、ROS、TNF-α、IL-1β、CRP、JNK、p38 MAPK mRNA、JNK、p38 MAPK升高(P<0.05);与模型组比较,药物对照组、黄芪甲苷组SOD、CTA升高,NSE、S100β、MDA、ROS、TNF-α、IL-1β、CRP、JNK、p38 MAPK mRNA、JNK、p38 MAPK降低,(P<0.05);与药物对照组比较,黄芪甲苷组SOD、CTA升高,NSE、S100β、MDA、ROS、TNF-α、IL-1β、CRP、JNK、p38 MAPK mRNA、JNK、p38 MAPK降低(P<0.05)。结论采用黄芪甲苷对急性脑梗死模型大鼠干预,可改善大鼠氧化应激指标,降低炎性反应,使大鼠神经功能得到改善,其作用机制可能与调控JNK/p38 MAPK信号通路有关。

共载黄芪甲苷-黄连素/白及多糖微球的制备与性能研究

利用白及多糖的生物黏附性制备共载黄芪甲苷-黄连素/白及多糖微球(AS-IV-BBR/BSP-Ms),并对其性能进行评价。结果表明,采用乳化交联法制备得到共载AS-IV-BBR/BSP-Ms,微球粒径在30~40μm,Zeta电位为-21.4 mV,且30 min内溶胀率达54.3%;红外光谱分析、X-射线衍射分析表明,黄芪甲苷和黄连素被包裹于白及多糖微球内部;DSC分析表明,在t=121℃时,出现尖锐吸热峰,热稳定性较好;体外抗氧化活性实验表明,该微球DPPH自由基的半数清除浓度(IC_(50))为8.23 mg/mL。该制备工艺稳定可行,微球性能评价达到预期目标,可为下一步药效学评价提供实验依据。

黄芪甲苷对慢性间歇性缺氧诱导的遗尿症大鼠TLR4-p38 MAPK通路的影响

目的 探讨黄芪甲苷对慢性间歇性缺氧诱导的遗尿症大鼠Toll样受体4(Toll-like receptor-4,TLR4)/p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase, p38 MAPK)信号通路的影响。方法 将SD大鼠随机分为对照组、模型组、黄芪甲苷低剂量组、黄芪甲苷中剂量组、黄芪甲苷高剂量组、去氨加压素组、脂多糖组、黄芪甲苷高剂量+脂多糖(Lipopolysaccharide, LPS)组,每组12只。除对照组外,其他组均需采用慢性间歇性缺氧诱导的方法构建遗尿症大鼠模型,各组大鼠每天于进入动物舱前1 h进行给药处理,1次/d,持续4周。末次处理24 h后,检测各组大鼠24 h排尿量及膀胱漏尿点压(bladder leak point pressures, BLPP)值的变化;ELISA法检测大鼠血清中抗利尿激素(antidiuretic hormone, ADH)含量;HE染色检测大鼠膀胱组织病理变化;比色法检测大鼠膀胱组织中超氧化物歧化酶(superoxide dismutase, SOD)活性、丙二醛(malonic dialdehyde, MDA)含量;Western blot检测大鼠膀胱组织中P2X3、TLR4、p-p38蛋白表达。结果 与对照组比较,模型组大鼠膀胱组织病理损伤严重,24 h排尿量增多,BLPP值变小,ADH含量、SOD活性降低,MDA含量和P2X3、TLR4、p-p38蛋白表达升高(P<0.05);与模型组比较,黄芪甲苷低剂量组、黄芪甲苷中剂量组、黄芪甲苷高剂量组、去氨加压素组大鼠膀胱组织病理损伤减轻,24 h排尿量减少,BLPP值变大,ADH含量、SOD活性升高,MDA含量和P2X3、TLR4、p-p38蛋白表达降低,LPS组大鼠膀胱组织病理损伤加剧,24 h排尿量增多,BLPP值变小,ADH含量、SOD活性降低,MDA含量和P2X3、TLR4、p-p38蛋白表达升高(P<0.05);与黄芪甲苷高剂量组比较,黄芪甲苷高剂量+LPS组大鼠膀胱组织病理损伤严重,24 h排尿量增多,BLPP值变小,ADH含量、SOD活性降低,MDA含量和P2X3、TLR4、p-p38蛋白表达升高(P<0.05)。结论 黄芪甲苷可能通过抑制TLR4/p38MAPK信号通路改善慢性间歇性缺氧诱导的大鼠遗尿症。

黄芪甲苷经由miR-125a-5p/NLRP1轴减轻椎间盘突出髓核细胞损伤

目的在白介素-1β(IL-1β)诱导退变的人髓核细胞中,探究黄芪甲苷对miR-125a-5p及其靶基因介导的信号通路的作用,揭示黄芪甲苷(astragaloside IV,AS-IV)对髓核细胞增殖、凋亡以及炎症反应的影响。方法实时荧光定量PCR(RT-qPCR)检测miR-125a-5p和核苷酸寡聚化结构域(NOD)样受体蛋白1(nucleotide oligomerization domain(NOD)-like receptor protein 1,NLRP1)在椎间盘突出患者髓核组织中的表达,用IL-1β诱导髓核细胞退变,在20、50和80μg·mL^(-1)黄芪甲苷干预浓度下检测miR-125a-5p和NLRP1的表达,在IL-1β和80μg·mL^(-1)黄芪甲苷处理的髓核细胞中单独或共同转染miR-125a-5p模拟物和NLRP1过表达质粒,然后分别检测细胞增殖、凋亡和炎症因子分泌情况,蛋白质免疫印迹(Western blot)检测细胞中信号通路相关蛋白p56和p38的磷酸化水平。结果椎间盘突出(lumbar disc herniation,LDH)患者的髓核组织中miR-125a-5p表达下调,NLRP1表达上调。黄芪甲苷促进IL-1β处理的髓核细胞中miR-125a-5p表达,减少NLRP1表达以及p56和p38蛋白的磷酸化水平。黄芪甲苷促进髓核细胞增殖,减少凋亡和炎症反应。结论黄芪甲苷通过上调miR-125a-5p表达,抑制NLRP1的表达和NF-κB/MAPK信号通路,减少IL-1β诱导的髓核细胞损伤。

黄芪甲苷对人外周血单个核细胞内HBV表达的影响

目的:探索黄芪甲苷对乙肝后肝硬化患者外周血单个核细胞(PBMC)内HBV表达的影响。方法:选取2019年2月—2022年2月于江西省人民医院就诊的20例乙肝后肝硬化患者,分离并培养外周血单个核细胞,依据培养液所含药物类型分为对照组、拉米夫定组、黄芪甲苷组。拉米夫定组和黄芪甲苷组又根据药物终浓度各分为低、中、高浓度3个亚组。培养1周后,分别检测各组HBsAg、HBeAg、HBV-DNA、HBV-cccDNA表达。结果:与对照组相比,黄芪甲苷各浓度组HBsAg表达量均显著降低,差异具有统计学意义(P<0.05);黄芪甲苷各浓度组HBeAg表达量无统计学差异(P>0.05),黄芪甲苷低浓度组HBV-DNA表达量无统计学差异(P>0.05),黄芪甲苷中、高浓度组HBV-DNA表达量显著降低,差异具有统计学意义(P<0.05),黄芪甲苷各浓度组HBV-cccDNA表达量无统计学差异(P>0.05)。与对照组相比,拉米夫定各浓度组HBsAg、HBeAg、HBV-DNA表达量均显著降低,差异具有统计学意义(P<0.05);拉米夫定低浓度组HBV-cccDNA表达量无统计学差异(P>0.05),拉米夫定中、高浓度组HBV-cccDNA表达量显著降低,差异具有统计学意义(P<0.05)。Spearman检验分析提示黄芪甲苷浓度与HBsAg、HBeAg、HBV-DNA、HBV-cccDNA表达量呈线性负相关,其中与HBsAg、HBV-DNA的负相关性有统计学意义(P<0.05)。结论:黄芪甲苷能够在一定程度上抑制乙肝后肝硬化患者PBMC中HBV的复制,能为解决肝移植术后HBV复发提供新的治疗思路。

黄芪甲苷对实验性自身免疫性脑脊髓炎小鼠T细胞免疫调节的影响

背景:多发性硬化初始阶段,中枢免疫细胞激活并释放大量炎症因子,引起白质脱髓鞘甚至累及灰质神经元。CD4^(+)T细胞不同亚群之间的分化平衡在实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的病程进展中发挥着重要作用。课题组前期研究结果表明黄芪内有效成分黄芪甲苷能够调节EAE小鼠体内免疫反应,其是否对T细胞亚群分化具有调节作用尚未明确。目的:探究黄芪甲苷对EAE小鼠治疗效果及其对T细胞的免疫调控机制。方法:将C57BL/6雌性小鼠分为正常对照组、EAE疾病模型组和黄芪甲苷治疗组,每组8只,后2组使用髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)制备EAE模型,免疫后第10-28天,黄芪甲苷治疗组以40 mg/(kg·d)灌胃给药。免疫当天至第28天,记录各组小鼠的体质量及临床评分;免疫后第28天取小鼠脊髓制成冰冻切片行苏木精-伊红染色、固蓝染色观察脊髓病理改变,流式细胞术检测脾脏T细胞亚群百分比,Western blot法检测脊髓组织中γ干扰素、白细胞介素17、白细胞介素6的蛋白表达,ELISA检测脾细胞上清液中γ干扰素、白细胞介素17、白细胞介素6、白细胞介素4水平。结果与结论:(1)与EAE疾病模型组相比,黄芪甲苷治疗能够减少EAE小鼠体质量丢失(P<0.05),缓解临床症状(P<0.05),减轻脊髓炎症细胞浸润及髓鞘脱失病理改变(分别为P<0.01和P<0.05);(2)与EAE疾病模型组相比,黄芪甲苷治疗可抑制表达γ干扰素和白细胞介素17的CD4^(+)T细胞亚群比例(分别为P<0.001和P<0.001),上调表达白细胞介素10和转化生长因子β的CD4^(+)T细胞亚群百分比(分别为P<0.001和P<0.01);(3)黄芪甲苷可下调脊髓和脾脏中γ干扰素(分别为P<0.05和P<0.01)、白细胞介素17(分别为P<0.05和P<0.05)、白细胞介素6(分别为P<0.05和P<0.05)的表达,上调脾脏中抑炎因子白细胞介素4的表达(P<0.01);(4)结果说明,黄芪甲苷可以减轻EAE小鼠的临床症状,其机制与调节脾脏免疫细胞亚群进而抑制炎症细胞向中枢浸润、减少髓鞘脱失有关。

黄芪甲苷防治心肌纤维化研究进展

心肌纤维化(myocardial fibrosis,MF)是指心脏成纤维细胞过度增殖、细胞外基质(extracellular matrix,ECM)过度堆积为主要表现的疾病,是多种心血管疾病发展到一定阶段的共同病理改变,表现为心肌重构及心肌代谢紊乱,最终导致心力衰竭而引发死亡。黄芪甲苷具有抗炎、抗氧化应激、抗细胞凋亡、改善心肌缺血再灌注等药理作用,在心肌纤维化防治方面有着良好的潜力。文章旨在对黄芪甲苷防治心肌纤维化的机制进行综述,为黄芪甲苷临床治疗心肌纤维化提供参考。

黄芪甲苷调控Nrf2/HO-1信号通路对血管内皮细胞氧化损伤的影响

目的 基于细胞实验研究黄芪甲苷(astragalosideⅣ,AST-Ⅳ)调控核转录因子红系2相关因子2/血红素加氧酶-1(nuclear factor-erythroid 2-related factor 2/heme oxygenase-1,Nrf2/HO-1)信号通路对血管内皮氧化损伤的影响。方法 采用1-棕榈酰基-2-(5'-氧-戊酰基)-sn-甘油-3-磷酸胆碱[1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine,POVPC]刺激血管内皮细胞24 h建立血管内皮细胞氧化损伤模型,随机分为模型(Model)组(35μmol/L POVPC)、AST-Ⅳ低剂量(AST-L)组(35μmol/L POVPC+50μmol/L AST-Ⅳ)、AST-Ⅳ高剂量(AST-H)组(35μmol/L POVPC+100μmol/L AST-Ⅳ)及AST-Ⅳ高剂量+ML385(Nrf2抑制剂)(AST-H+ML385)组(35μmol/L POVPC+100μmol/L AST-Ⅳ+5μmol/L ML385),以正常未处理细胞作为对照(Control)组。免疫荧光鉴定大鼠胸主动脉内皮细胞(aortic vascular endothelium cell,AVEC),CCK-8检测AST-Ⅳ细胞增殖情况,Transwell小室检测AVEC迁移情况,Matrigel基质胶检测细胞血管形成功能,鬼笔环肽染色检测细胞骨架结构,DCFH-DA荧光探针检测细胞活性氧(reactive oxygen species,ROS)水平,试剂盒法检测细胞培养上清液中超氧化物歧化酶(superoxide dismutase,SOD)水平,RT-qPCR和Western blot检测Nrf2和HO-1的mRNA及蛋白表达情况。结果 免疫荧光鉴定所提取的细胞,可特异性表达血管性血友病因子(von willebrand factor,v WF),鉴定为AVEC。与Control组相比,Model组细胞活力、迁移能力和血管形成功能显著下降(P<0.01),细胞骨架破坏明显,细胞内ROS水平显著上升(P<0.01),细胞培养上清液中SOD含量显著减少(P<0.01),细胞中Nrf2和HO-1的m RNA及蛋白表达水平下调(P<0.01或P<0.05)。与Model组比较,AST-L组及AST-H组细胞活力、迁移能力和血管形成功能显著升高(P<0.01),细胞骨架破坏得到较好修复,细胞内ROS水平显著下降(P<0.01),细胞培养上清液中SOD含量显著增加(P<0.01),细胞中Nrf2和HO-1的mRNA及蛋白表达水平显著上调(P<0.01),且以AST-H组效果更为显著(P<0.01)。与AST-H组比较,ASTH+ML385组迁移能力和血管形成功能显著下降(P<0.01),细胞骨架破坏增加,细胞内ROS水平显著上升(P<0.01),细胞培养上清液中SOD含量显著减少(P<0.01),细胞中Nrf2和HO-1的mRNA及蛋白表达水平显著下调(P<0.01)。结论 AST对血管内皮氧化损伤具有保护作用,且有一定的剂量依赖性,其机制可能是通过激活Nrf2/HO-1信号通路发挥作用。

徐黄合剂的制备工艺优化及含量测定

优化徐黄合剂最佳提取工艺并建立制剂含量测定方法。以黄芪甲苷含量为提取工艺及含量测定评价指标,采用L_(9)(3^(4))正交试验法,选取加水量、煎煮时间及煎煮次数为考察因素确定最佳提取工艺,并考察常压浓缩和减压浓缩对黄芪甲苷含量的影响。另外建立黄芪甲苷高效液相色谱测定方法并进行方法学考察。结果显示,最佳提取工艺为加6倍量水,沸后提取1 h,提取2次;两种不同的浓缩方法对制剂含量影响无显著性差异;黄芪甲苷在0.048 16~0.963 20 mg/mL范围内线性关系良好,阴性样品无干扰,精密度、重复性、稳定性测得的RSD<3%;平均加样回收率为101.85%(RSD=2.54%,n=9),表明该含量测定方法灵敏、准确。

淫黄葛合剂对APP/PS1 HAMP^(-/-)小鼠认知功能和铁死亡氨基酸代谢通路的影响

目的:探究淫羊藿苷-黄芪甲苷-葛根素(淫黄葛)合剂(YHG)对铁调素(hepcidin,HAMP)基因敲除的APPswe/PS1dE9(APP/PS1 HAMP^(−/−))小鼠认知功能及铁死亡氨基酸代谢通路的影响。方法:实验分为阴性对照(C57BL/6小鼠)组、APP/PS1组、APP/PS1 HAMP^(−/−)组、APP/PS1+YHG组、APP/PS1 HAMP^(−/−)+YHG组、APP/PS1+地拉罗司(DFX)组和APP/PS1 HAMP^(−/−)+DFX组,每组6只。YHG(淫羊藿苷、黄芪甲苷和葛根素的剂量分别为120、80和80 mg/kg)和DFX(100 mg/kg)灌胃给药,C57组、APP/PS1组和APP/PS1 HAMP^(−/−)组小鼠给予等体积蒸馏水灌胃,每日一次,用药2个月。采用组织铁试剂盒检测小鼠脑组织铁离子含量;透射电镜检测小鼠海马神经元线粒体形态学变化;Morris水迷宫检测小鼠学习记忆能力;免疫荧光法检测各组小鼠脑组织神经元核抗原(NeuN)的含量;生化试剂盒检测小鼠脑组织谷胱甘肽(GSH)表达情况;Western blot检测小鼠脑组织谷氨酸半胱氨酸连接酶催化亚基(GCLC)和谷氨酰胺酶2(GLS2)的表达情况。结果:与C57BL/6小鼠相比,APP/PS1小鼠脑铁含量显著升高(P<0.01),线粒体形态破坏严重,学习记忆能力显著下降(P<0.05),脑组织神经元损伤严重(P<0.01),GSH、GCLC和GLS2表达水平均显著降低(P<0.01);与APP/PS1小鼠相比,APP/PS1 HAMP^(−/−)小鼠脑铁含量显著升高(P<0.01),线粒体形态破坏严重,学习记忆能力显著下降(P<0.05),脑组织神经元损伤严重(P<0.01),GSH、GCLC和GLS2表达水平显著降低(P<0.05);使用YHG和DFX后各组小鼠脑铁含量显著降低(P<0.01),线粒体形态破坏减轻,学习记忆能力显著升高(P<0.05),脑组织神经元损伤减轻(P<0.01),GSH、GCLC和GLS2表达水平显著升高(P<0.05)。结论:YHG可改善APP/PS1 HAMP^(−/−)小鼠学习记忆能力,其机制可能与调节铁死亡氨基酸代谢、增强抗氧化能力有关。