Beta-asarone protects against MPTP-induced Parkinson’s disease via regulating long non-coding RNA MALAT1 and inhibiting α-synuclein protein expression

ObjectiveNumerous long non-coding RNAs (lncRNA) have been identified in neurodegenerative disorders including Parkinson’s disease (PD). Emerging evidence demonstrates that β-asarone functions as neuroprotective effects in both in vitro and in vivo models. However, the role of β-asarone and its potential mechanism in PD remain not completely clear.MethodsMPTP-induced PD mouse model and SH-SY5Y cells subjected to MPP+ as its in vitro model were used to evaluate the effects of β-asarone on PD. LncRNA MALAT1 and α-synuclein expression were determined by real-time PCR and western blot methods.Resultsβ-Asarone significantly increased the TH+ cells number and decreased the expression levels of MALAT1 and α-synuclein in midbrain tissue of PD mice. RNA pull-down and immunoprecipitation assays confirmed that MALAT1 associated with α-synuclein, leading to the increased stability of α-synuclein and its expression in SH-SY5Y cells. β-asarone elevated the viability of cells exposed to MPP+. Either overexpressed MALAT1 or α-synuclein could canceled the protective effect of β-asarone on cell viability. In PD mice, pcDNA-MALAT1 also decreased the TH+ cells number and increased the α-synuclein expression in PD mice with treatment of β-asarone.Conclusionβ-Asarone functions as a neuroprotective effect in both in vivo and in vitro models of PD via regulating MALAT1 and α-synuclein expression.     

基于灰色关联度分析的丹参提取物抗炎作用谱效关系研究

目的研究丹参不同提取物的超高效液相色谱(UPLC)图谱与抗炎作用的谱效关系,为明确丹参抗炎作用物质基础提供依据。方法采用UPLC法对丹参不同提取部位(S1~S9)进行分析,以人支气管上皮细胞炎症模型考察不同提取部位的抗炎作用,采用灰色关联度分析法建立其谱效关系。结果丹参醇提物50%乙醇洗脱液(S3)、80%乙醇洗脱液(S4)、滤过沉淀(S5)以及萃取后水层提取物(S9)对于炎症细胞抗炎能力强,其中以丹参醇提物的50%乙醇洗脱液(S3)作用能力最强。丹参发挥抗炎作用的药效是多种成分共同作用的结果,各特征峰所代表的化学成分对其抗炎药效贡献的大小顺序为9号峰1号峰2号峰6号峰15号峰11号峰12号峰7号峰10号峰5号峰14号峰3号峰18号峰4号峰13号峰16号峰8号峰,对抗炎作用药效贡献大的前4个峰分别为9、1、2、6号峰,经过与对照品比对及质谱分析鉴定出4个峰对应的成分分别为丹参酮IIA、丹参素、丹酚酸B和3′-甲基丹酚酸B。结论丹参抗炎作用的药效是多种成分共同作用的结果,明确了丹参中丹参酮IIA、丹参素、丹酚酸B和3′-甲基丹酚酸B这4个成分对其抗炎作用的药效贡献最大,为今后探索丹参药材抗炎作用的药效物质基础提供了一种新思路。     

赤芍中氧化芍药苷和芍药苷的含量测定

目的:建立同时测定赤芍中氧化芍药苷和芍药苷含量的HPLC方法。方法:采用反向高效液相色谱法,使用AglientZorbaxSB-C18(4.6mm×250mm,5μm)色谱柱,流动相为乙腈—水,流速1mL/min,检测波长254nm,柱温35℃,进样量20μL。结果:氧化芍药苷回归方程为Y=41607X-164942,R=0.9994,线性范围为0.02525～0.20200mg/mL;芍药苷回归方程为Y=3641.6X+174622,R=0.9995,线性范围为0.2525～2.0200mg/mL。结论:建立了一个测定赤芍中氧化芍药苷和芍药苷含量的方法。     

Immunology: Paper alert

A selection of interesting papers that were published in the two months before our press date in major journals most likely to report significant results in immunology.     

A New Alkaloid from Bombycis Feculae and Its α-Glucosidase Inhibitory Activity

Objective To study the chemical constituents of Bombycis Feculae.Methods Chemical constituents were isolated by HPLC-ELSD.The structures of the isolated compounds were determined by spectral means.Results Two compounds were isolated and identified as 1-deoxynojirimycin(1)and(2R,3R,5R)-2-(hydroxymethyl) piperidine-3,5-diol,named as 1,3-dideoxygalatonojirimycin(2).Conclusion Compound 2 is a new alkaloid.The extract of Bombycis Feculae,compound 1 and compound 2 show inhibitory activities againstα-glucosidase.     

大黄对瘢痕成纤维细胞增殖抑制作用及其有效成分研究

探索中药大黄抗瘢痕成纤维细胞(HSF)增殖作用的有效成分。采用索氏提取法将大黄药材按照极性依次提取不同的部位;以MTS法筛选大黄各溶剂提取物(25μg·mL-1)抗人增生性瘢痕成纤维细胞增殖的作用,结合流式细胞仪检测其对细胞周期的影响;进一步采用HPLC对其有效成分进行谱效分析,将其中高活性成分进行UPLC-Q/TOF鉴定及验证。结果显示,大黄乙酸乙酯提取物的有较高的抗细胞增殖活性(P<0.01),能显著增加G0/G1期细胞的比例(P<0.01),降低增殖指数(PI)(P<0.01),其中主要活性成分为蒽醌类化合物;以大黄素、大黄酸和没食子酸为例的验证结果显示,这3种成分均能抑制细胞的增殖且呈剂量依赖性。结果提示,大黄乙酸乙酯提取物对瘢痕成纤维细胞增殖有抑制作用,主要成分为其中的蒽醌类化合物。     

HPLC、UPLC测定芪参益气滴丸中5个丹参活性成分含量的比较研究

目的:建立同时测定芪参益气滴丸中5个丹参活性成分(丹参素、原儿茶醛、迷迭香酸、丹酚酸B、丹酚酸A)含量的高效液相色谱(HPLC)和超高效液相色谱(UPLC)方法,并对建立的方法进行比较。方法:HPLC方法色谱柱为Agilent Zorbax SBC18柱(4.6 mm×250 mm,5μm),UPLC方法色谱柱为Waters Acquity BEH C18柱(2.1 mm×100 mm,1.7μm),流动相均为乙腈-甲酸-水体系梯度洗脱,2种方法的流速分别为1.0和0.4 mL.min-1,检测波长均为280 nm,柱温均为30℃。比较2种方法的分离情况及线性、精密度、重复性、加样回收率、样品测定结果。结果:UPLC的分析时间少于HPLC;5个物质在一定范围内均呈现良好线性,r均大于0.9998;2种方法的精密度、重复性、加样回收率和样品测定结果无显著性差异。结论:HPLC和UPLC 2种方法准确性、重复性均较好;UPLC分析比HPLC具有更加快速、高效的优势。     

远志蔗糖酯类新化学成分的分离与鉴定

目的对远志根的化学成分进行研究。方法采用反复硅胶柱色谱、Sephadex LH-20柱色谱和半微量制备高效液相色谱分离纯化,根据ESI-MS、1H-NMR和13C-NMR等谱学数据进行结构鉴定。结果从远志根的乙醇提取物中分离得到11个单体化合物,分别鉴定为蔗糖酯类化合物西伯利亚远志糖A5(sibiricose A5,1)、tenuifolioside A(α-D-glucopyranoside,3-O-[(2Z)-3-(4-hydroxy-3-me-thoxyphenyl)-1-oxo-2-propen-1-yl]-β-D-fructofuranosyl,2)、tenuifolioside B(α-D-glucopyranoside,3-O-[(2E)-3-(3,4-dimethoxyphenyl)-1-oxo-2-propen-1-yl]-β-D-fructofuranosyl,3)、tenuifolioside C(α-D-glucopyranoside,3-O-[(2Z)-3-(3,4-dimethoxyphenyl)-1-oxo-2-propen-1-yl]-β-D-fructofurano-syl,4)、西伯利亚远志糖A6(sibiricose A6,5)、球腺糖A(glomeratose A,6);呫吨酮类化合物西伯利亚远志呫吨酮B(sibiricaxanthone B,7)、远志呫吨酮Ⅺ(polygalaxanthoneⅪ,8)、1,2,3,7-四甲氧基呫吨酮(1,2,3,7-tetramethoxyxanthone,9)和远志酮Ⅱ(onjixanthoneⅡ,10);远志醇(polygitol,11)。结论化合物2~4为首次发现的新化合物。