基于网络药理学探讨石菖蒲有效成分治疗神经退行性疾病的研究

目的基于网络药理学分析石菖蒲主要成分治疗神经退行性疾病(neurodegenerative diseases,ND)的作用靶点和机制。方法利用TCMSP(traditional Chinese medicine systems pharmacology database and analysis platform)数据库检索和既往文献证明得到石菖蒲的主要有效成分。在GeneCards,OMIM(Online Mendelian Inheritance in Man),TTD(Therapeutic Target Database)数据库检索ND的相关靶点,并用Uniprot数据库进行标准化。构建ND和石菖蒲有效成分的共有靶点韦恩图和PPI网络图,进行GO(Gene ontology)注释和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,展示石菖蒲有效成分在治疗ND过程中疾病-靶点的关系。结果石菖蒲和ND的共有靶点总计为41个,关键靶点主要有FOS、ACHE、SLC6A4、ADRA1B、ADRA2A、MAOB、PTGS2、ADRA1A等。这些靶点涉及G蛋白偶联的胺受体活性、肾上腺素受体活性、儿茶酚胺结合、类固醇激素受体活性、神经递质受体活性、突触后神经递质受体活性等与突触间神经递质及其受体密切相关的生物过程、钙信号通路、cGMP-PKG信号通路和唾液分泌通路等信号通路。结论石菖蒲的主要有效成分可能通过对FOS、ACHE、SLC6A4、ADRA1B、MAOB等蛋白及相关通路的影响进而对神经神经递质受体或突触的损伤修复发挥治疗ND的作用,这为石菖蒲有效成分治疗ND的作用机制提供了新方向。

β-细辛醚激活PINK1/Parkin介导的线粒体自噬改善APP/PS1小鼠认知和记忆功能障碍的研究

目的探讨β-细辛醚对淀粉样前体蛋白(APP)/早老素1(PS1)小鼠的作用机制及对PINK1/Parkin介导的线粒体自噬的影响。方法将APP/PS1小鼠随机分为模型组、β-细辛醚低、中、高剂量组(15、30、45 mg/kg)、多奈哌齐组(1 mg/kg)、雷帕霉素组(4 mg/kg)、3-甲基腺嘌呤(3-MA)组(4 mg/kg)、环孢霉素A组(10 mg/kg),另设正常组,每组10只。各组灌胃给药,模型组和正常组用等体积蒸馏水代替,1次/d,给药30 d。观察各组小鼠水迷宫实验,蛋白质印迹法检测小鼠海马中APP、PS1、神经突触素1(SYN1)、β-位点淀粉样前体蛋白裂解酶1(BACE1)、LC3 I/II、Beclin-1、p62、PINK1、Parkin和p-Parkin蛋白表达水平;反转录聚合酶链反应检测APP、PS1、BACE1、LC3B、Beclin-1、p62和Parkin mRNA表达水平。结果与模型组相比,β-细辛醚高剂量组、β-细辛醚中剂量组和多奈哌齐组APP/PS1小鼠的逃避潜伏期缩短(P<0.05或P<0.01),小鼠海马内APP、PS1、BACE1的表达下降(P<0.01),SYN1表达上升(P<0.05或P<0.01);与模型组相比,β-细辛醚中剂量组APP/PS1小鼠内Beclin-1、LC3B表达均升高(P<0.05或P<0.01),p62表达下降(P<0.05);与模型组相比,雷帕霉素组Beclin-1、LC3B表达均升高(P<0.05或P<0.01),p62表达下降(P<0.01);与模型组相比,3-MA组p62表达升高(P<0.05);与模型组相比,β-细辛醚中剂量组APP/PS1小鼠内PINK1、p-Parkin表达均显著升高(P<0.01)。结论β-细辛醚通过激活PINK1-Parkin介导的线粒体自噬而影响阿尔茨海默病病理,这为应用β-细辛醚治疗阿尔茨海默病提供了新的靶点。

β-细辛醚治疗帕金森病抑郁的网络药理学作用机制分析

目的基于网络药理学分析β-细辛醚治疗帕金森病抑郁(DPD)的作用机制。方法利用TCMSP、Swiss Target Prediciton和PharmMapper数据库检索得到β-细辛醚的作用靶点,通过Genecard数据库和OMIM数据库检索DPD的疾病相关靶点,并通过UniProt数据库进行标准化。构建β-细辛醚和DPD的共有靶点韦恩图,对共有靶点进行蛋白互作(PPI)网络分析,以获取关键靶点信息;对共有靶点进行GO功能及KEGG通路富集分析;建立"药物-疾病-靶点-通路"网络,展示β-细辛醚治疗DPD过程中疾病-靶点-通路的关系。结果共得到β-细辛醚和DPD的共有靶点64个,包括ALB、c-FOS、EGFR、JUN、CASP3、PTGS2、IGF1、ESR1、APP等多个关键靶点。共有靶点涉及学习或记忆、突触后膜的内在成分、神经递质受体活性、突触后神经递质受体活动等与突触间神经递质及其受体密切相关的生物过程、分子功能及细胞组成,以及5-HT能神经突触、DA能神经突触和神经活性配体受体相互作用等信号通路。结论β-细辛醚可能通过对c-FOS、JUN、ALB、EGFR、CASP3、MAP2K1、MAPK14等靶点及相关通路的作用,进而影响5-HT能神经突触和DA能神经突触的损伤修复,发挥治疗DPD的作用。

Gene expression profiling under different photoperiod/temperature conditions in a photoperiod-/thermo-sensitive genic male sterile line of rice(Oryza sativa L.)

Peiai 64S is a photoperiod- and thermo- sensitive genic male sterile (PTSGMS) line of rice, which is male sterile at long day/high temperature and partial fertile at short day/low temperature. A cDNA array representing 3328 unique rice genes was used to profile the gene expression patterns in the young panicles of Peiai 64S under these two condi- tions. The statistical data showed that up to 14.60% of genes exhibited up-or down-regulated expressions in the plants at long day/high temperature compared with plants at short day/low temperature. Only four genes were up-regulated while 482 genes down-regulated. Real-time PCR with all the up-regulated and 9 randomly selected down-regulated genes confirmed the differential expressions detected by the array, indicating that the constructed cDNA array is reliable. These differently expressed genes participated in almost all cell biological responses. Analysis of up- and down-regulated genes revealed distinctive changes between the mRNA abundances of MMK1 and MMK2, both of which are analogs ofMAPK, and significant down-regulation of several transcription factors. It was suggested that changes that occurred in the MAPK signal transduction path- way might disturb the transcription control necessary for morphogenesis of pollens and corresponding phys- iological functions.

Multiple splicing types of OsRIX4,an RAD21 homolog in rice (Oryza sativa L.)

The present paper describes multiple splicing types of OsRIX4,an RAD21 homolog in rice. A type of alternative splicing (AS),distinctive from all five previously known splicing types,was identified in which interior sequences of a constitutive exon could be spliced. Translation of the transcript produced with this AS type was demonstrated at the protein level. Expression of multiple transcripts was organ specific. The expression abundance of transcripts,OsRIX4-4 and OsRIX4-5,was positively correlated with fertility in rice. The splicing type identified in the present study provided the means to further un-derstand and define different mRNA splicing types in plants and suggested that post-transcription processing of mRNA and its regulation mechanism are complex.