Network pharmacology and verification experiment-based prediction of active components and potential targets of Alpiniae Oxyphyllae Fructus-Saposhnikoviae Radix(Yizhiren-Fangfeng)for treatment of diabetic kidney disease

Background:In this study,we analyzed the potential active components,related crucial targets and possible signaling pathway mechanisms of Alpiniae Oxyphyllae Fructus and Saposhnikoviae Radix(AOF-SR)herb pairs in the treatment of diabetic kidney disease(DKD)using network pharmacology and verification experiments.Methods:The active compounds and potential targets of AOF-SR were derived from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,The Encyclopedia of Traditional Chinese Medicine,and PubChem databases,and the potential therapeutic targets of DKD were derived from the OMIM,Drugbank,and DisGeNET databases.The“compounds-diseases-targets”network was constructed using Cytoscape 3.6.0.ClusterMaker functionality in Cytoscape is being used to screen important targets for AOF-SR treatment of DKD.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of important targets were performed using DAVID database.In addition,according to the predicted results of network pharmacology,HK-2 cells were used to construct DKD model for verification experiment.HK-2 cells were divided into control group,high glucose(HG)group and AOF-SR(HG+AOF-SR)group to detect survival rate and expression of key proteins in NF-κB and PI3K/Akt signaling pathways.Results:A total of 38 compounds were selected from AOF-SR,of which 23 were Alpiniae Oxyphyllae Fructus and 15 were Saposhnikoviae Radix.Through enrichment analysis of 82 important targets,88 signaling pathways were identified;some of these pathways,such as the NF-κB,PI3K-Akt,IL-17,and JAK/STAT signaling pathways,regulate the pathological process of DKD.In verification experiment,the HK-2 cells survival rate was higher in the HG+AOF-SR group than in the HG group(P<0.05).Moreover,western blotting results showed that the expression levels of NF-κB,p-PI3K,and p-Akt in HG+AOF-SR group were significantly lower than those in HG group(P<0.05).Conclusion:Overall,this study revealed the active compounds,important targets and possible mechanisms of AOF-SR treatment for DKD,and conducted preliminary verification experiments on its correctness,provided novel insights into the treatment of DKD by AOF-SR.

益智GAP栽培的环境评价

为制定益智(Alpinia oxyphylla Miq)SOP和建设益智GAP种植基地提供科学依据,对海南省益智主要生产区及野生生长区环境包括土壤、水体及大气进行采样综合分析评价。结果表明,益智GAP种植基地——海南省琼中长兴村规划区大气环境达到国家第一级标准要求,灌溉水资源达到国家标准要求,土壤环境达到国家第二级标准要求。海南益智GAP种植区环境质量良好,符合绿色药材栽培的环境质量要求。

野生益智的光合特性研究

以陵水、保亭、万宁3地的野生益智(Alpinia oxyphylla Miq.)为材料,研究不同地点益智的光合特性,包括净光合速率、气孔导度、胞间CO2浓度、蒸腾速率、叶面积、叶绿素含量、土壤养分及它们之间的关系。结果表明,除保亭和万宁的气孔导度没有显著差异外,其他光合指标都存在显著差异;3地的叶面积存在显著差异,保亭和万宁、陵水和万宁的叶绿素含量也存在显著差异;陵水与万宁、保亭与万宁的土壤氮、磷含量差异都达到了显著水平,保亭与陵水、万宁与陵水的土壤钾含量差异达到了显著水平;净光合速率与叶面积、叶绿素含量呈显著正相关,胞间CO2浓度与叶绿素含量呈极显著正相关;土壤磷含量与益智的高度和密度呈显著或极显著正相关,与净光合速率呈显著负相关,与胞间CO2浓度呈极显著正相关;钾含量与净光合速率、蒸腾速率呈极显著负相关;氮含量与气孔导度、蒸腾速率呈显著正相关。试验结果为提高益智的品质,为野生益智的驯化、改良和保护提供了基础资料与科学依据。

Oxyphyllones A and B,novel sesquiterpenes with an unusual 4,5-secoeudesmane skeleton from Alpinia oxyphylla

Two novel 4,5-secoeudesmane sesquiterpenoids, oxyphyllones A （1） and B （2） were isolated from the fruits of Alpinia oxyphylla. Their structures were established by spectroscopic methods including 1D and 21） NMR spectra. These two compounds are the first example of naturally occurring sesquiterpenoids with a 4,5-secoeudesmane skeleton in the family of Zingiberaceae and oxyphyllone A （1） is the first 4,5-secoeudesmane type of 13-norsesquiterpenoid. Compounds 1 and 2 exhibited no cytotoxicities against three cancer cell lines at 10 μg/mL. ？ 2009 Ning Hua Tan. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Protective effects of organic extracts of Alpinia oxyphylla against hydrogen peroxide-induced cvtotoxicitv in PC12 cells

Alpinia oxyphylla,a traditional herb,is widely used for its neuroprotective,antioxidant and memory-improving effects.However,the neuroprotective mechanisms of action of its active ingredients are unclear.In this study,we investigated the neuroprotective effects of various organic extracts of Alpinia oxyphylla on PC12 cells exposed to hydrogen peroxide-induced oxidative injury in vitro.Alpinia oxyphylla was extracted three times with 95%ethanol(representing extracts 1–3).The third 95%ethanol extract was dried and resuspended in water,and then extracted successively with petroleum ether,ethyl acetate and n-butanol(representing extracts 4–6).The cell counting kit-8 assay and microscopy were used to evaluate cell viability and observe the morphology of PC12 cells.The protective effect of the three ethanol extracts(at tested concentrations of 50,100 and 200μg/mL)against cytotoxicity to PC12 cells increased in a concentration-dependent manner.The ethyl acetate,petroleum ether and n-butanol extracts(each tested at 100,150 and 200μg/mL)had neuroprotective effects as well.The optimum effective concentration ranged from 50–200μg/mL,and the protective effect of the ethyl acetate extract was comparatively robust.These results demonstrate that organic extracts of Alpinia oxyphylla protect PC12 cells against apoptosis induced by hydrogen peroxide.Our findings should help identify the bioactive neuroprotective components in Alpinia oxyphylla.

Anti-infective, cytotoxic and antioxidant activity of Ziziphus oxyphylla and Cedrela serrata

Objective: To search for the most active antimicrobial and antioxidant sub-fractions related to traditional use of Ziziphus oxyphylla(Z. oxyphylla) and Cedrela serrata(C. serrata) in Pakistan against infectious and liver diseases.Methods: Factions of different polarity were tested in vitro for their antiprotozoal,antimalarial, antibacterial and antifungal activity against different pathogens. Cytotoxicity on MRC-5 cell lines(human lung fibroblasts) as well as, in vitro radical scavenging activity was evaluated using the 1, 1-diphenyl-2-picrylhydrazyl radical assay.Results: The highest antiprotozoal activity was observed for the CHCl3 fractions of Z. oxyphylla roots and leaves, and C. serrata bark. The CHCl3 and Et OAc fractions of Z. oxyphylla roots, the CHCl3 fraction of Z. oxyphylla leaves, the Et OAc and the residual Me OH: H2 O fraction of C. serrata bark showed antibacterial activity against Staphylococcus aureus. The same residual Me OH: H2 O fraction of C. serrata bark was active against Candida albicans. The highest antioxidant activity was observed for the more hydrophilic Et OAc fractions of Z. oxyphylla leaves, C. serrata bark and leaves, and the residual Me OH: H2 O fraction of C. serrata bark.Conclusions: This study supports at least in part the traditional uses of these plants for antimicrobial purposes and against liver diseases.

Effects of Different Harvest Periods on Seed Quality of Alpinia oxyphylla Miq.

[Objectives] To clarify the appropriate harvest period of Alpinia oxyphylla Miq. seed and improve the quality of seeds. [Methods] In this experiment,the basic parameters and germination characteristics of A. oxyphylla Miq. seeds during different harvest periods were analyzed and compared. [Results] The results showed that the A. oxyphylla Miq. fruit picking period was closely related to the seed quality,and the seed maturity was highest 100 d after the flowering of A. oxyphylla Miq.,that is,the A. oxyphylla Miq. fruit skin turned from green to yellow( yellow-green),the peel had a small number of brown spots,it was spicy enough,the fruit dry/fresh weight ratio was greater than0. 34,the seed thousand kernel weight reached 11 g or more,and the dried seed moisture content was 13. 01%. Using the TTC method,the seed viability index was determined to be 85%,and in this case,it was the best time of harvesting seed. The water permeability of A. oxyphylla Miq. seed was poor,and the seed imbibition and water absorption time was long,about 72 h. The maximum water absorption rate was about23. 71% at the imbibition and water absorption stage,and it was inferred that the A. oxyphylla Miq. seeds were hard seeds. The germination time of A. oxyphylla Miq. seeds was long,the mature A. oxyphylla Miq. seeds began to germinate on the 12 th d,and the germination rate reached 75. 56% on the 50 th d. [Conclusions] This study determined the optimum harvest period of A. oxyphylla Miq. seeds,clarified the biological characteristics of slow seed water absorption and long germination cycle,and concluded that the seed coat was the main factor leading to slow seed water absorption and long seed germination cycle.

Exploration of the effective components and mechanism of Alpinia oxyphylla-Saposhnikovia Radix in the treatment of diabetic kidney disease based on network pharmacology

Objective:Network pharmacology was used to predict the main targets and related mechanism pathways of the key compounds and active ingredients of the Alpinia oxyphylla-Saposhnikovia Radix based on the therapeutic strategy of tonifying kidney and dispelling pathogenic wind for the treatment of diabetic kidney disease(DKD).Methods:The effective components of Alpinia oxyphylla-Saposhnikovia Radix were screened through the data platform of Traditional Chinese Medicine(TCM).After screening effective targets,the disease target gene database was used to obtain targets for DKD,and then gene enrichment analysis to predict relevant signalling pathways after obtaining the key target genes through Venn analysis.Results:There were 324 active components of Alpinia oxyphylla-Saposhnikovia Radix through the data platform of TCM.In addition,Venn analysis showed that there were 48 key target genes for the treatment of DKD.The biological pathways for the action of Alpinia oxyphylla-Saposhnikovia Radix in DKD were closely related to many signaling pathways including NF-kappaB,classical MAP kinase,JNK and P38 MAP kinase,and ERK5.Conclusions:Treatment of DKD with Alpinia oxyphylla-Saposhnikovia Radix based on the therapeutic strategy of tonifying kidney and dispelling pathogenic wind has the characteristics of multi-components,multi-targets and multi-pathways,which could provide new ideas and methods for further researches on the treatment of DKD with Alpinia oxyphylla-Saposhnikovia Radix.

Effects of Alpinia oxyphylla on oxidative stress and expression of p47phox in diabetic nephropathy rats

Objective:To explore whether Yizhi(Alpinia oxyphylla,AO)could treat diabetic nephropathy(DN)through the inhibition of p47phox expression which is related to oxidative stress.Methods:Model of DN was established in SD rats with high glucose and fat diet for 4 weeks after STZ intraperitoneal injection.The rats were randomly divided into blank group,model group,AO group and irbesartan(IRB)group.Drugs were administered for 4 weeks.Blood glucose and 24-hour urinary protein were measured regularly.The index of ROS,SOD,MDA and Nox2 were tested by ELISA and the expression of p47phox protein in renal tissues were tested by techniques of RT-PCR and Western blot.Results:The decrease of blood sugar in AO group and IRB group were not obvious(P>0.05)while the drugs could significantly reduce the amount of protein in urine(P<0.05).AO could decrease ROS,MDA and Nox2 and increase SOD(P<0.05).It also could inhibit the expression of p47phox which was increased in model group(P<0.05).The IRB group has the similar effects compared with AO group(P>0.05).Conclusion:AO could reduce the amount of 24-hour urinary protein in DN rats and the mechanism may be related to the inhibition of p47phox expression thus alleviates excessive oxidative stress response.

益智炭疽病病原鉴定

炭疽病是南药植物益智上普遍发生的一种病害。为明确其病原类型,本研究通过形态观察、柯赫氏法则验证、ITS序列比对和系统进化分析等方法,对来自海南省5个市县的病样进行了病原鉴定。结果显示,18份样本中分离纯化到8个菌株,形态学上分为四类,均具有炭疽菌的典型特征。进一步结合8个菌株的ITS序列测定、NCBI数据库中的序列比对、序列之间多重比对和系统进化分析,将菌株YZ-HN-022鉴定为C.aenigma,YZ-HN-015-1、YZ-HN-001-1-2、YZ-HN-016-2、YZ-HN-010和YZ-HN-017鉴定为C.gloeosporioides复合种,YZ-HN-015-1与其他菌种的种内分化较明显;YZ-HN-020-1-1和YZ-HN-020-1-2鉴定为C.boninense。所有病原均为在益智上的首次报道。研究结果为进一步弄清益智炭疽病病原遗传分化、病害流行和防治技术等提供参考。

益智分子生药学研究进展

分子生药学是利用分子生物学技术对生药进行鉴定、保护、栽培和有效成分生产研究的一门新兴学科,它让生药的研究从微观向基因水平迈进。益智的药用价值和商业价值很高可入药,可食用。由于姜科植物的果实表型特征很多都极为相似,传统方法不易辨别,且益智在基因水平上的研究略显不足,故而益智的分子生药学研究逐渐得到了广大学者们的关注。本研究从近几年来益智的分子生药学方面的有关研究进展,对今后益智分子生药学的研究方向和重点给出了建议,以期能够为益智种质资源的保护生物学研究和开发、利用提供参考。

胡萝卜苷棕榈酸酯改善阿尔茨海默病大鼠学习记忆障碍的神经保护作用

阿尔茨海默病(Alzheimer’s disease,AD)是一种与年龄相关的神经退行性疾病,其特征是进行性记忆减退和认知障碍。淀粉样β蛋白(Aβ)被认为是AD发病的致病因素。越来越多的证据表明Aβ神经毒性是由谷氨酸兴奋毒性介导的。胡萝卜苷棕榈酸酯(daucosterol palmitate,DSP)是从抗衰老中草药益智(Alpinia oxyphylla)中分离得到的一种具有抗谷氨酸兴奋毒性作用的植物类固醇。在我们之前的研究中。基于DSP的抗谷氨酸兴奋毒性作用,探讨了DSP改善AD模型大鼠学习记忆障碍的潜在保护作用及其机制。结果表明,DSP能有效改善Aβ诱导的大鼠学习记忆障碍,显著抑制Aβ诱导的海马组织ROS产生,有效地防止Aβ诱导的海马神经元损伤,显著恢复海马突触素的表达水平。这项研究表明,DSP可能是一个开发作为AD认知下降的治疗剂潜在的候选。

益智种子繁殖育苗技术

主要阐述益智(Alpinia oxyphylla Miq.)种子育苗技术,主要包括选种、采种、种子处理、播种和苗期管理等技术措施。