Protein target identification of ginsenosides in skeletal muscle tissues: discovery of natural small-molecule activators of muscle-type creatine kinase

Chen, Feiyan,Zhu, Kexuan,Chen, Lin,Ouyang, Liufeng,Chen, Cuihua,Gu, Ling,Jiang, Yucui,Wang, Zhongli,Lin, Zixuan,Zhang, Qiang,Shao, Xiao,Dai, Jianguo,Zhao, Yunan The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.3

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, the mechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides) in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potential target in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides, had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol (PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in the study, was selected as a representative to confirm direct binding and its biological importance. Biolayer interferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPD specifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by molecular docking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activity in vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the function of the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delaying exercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginseng reduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can help in further developing better CK-MM activators based on the dammarane-type triterpenoid structure.

Fast Response Research of Magnetically Controlled Reactor

Chen Feng,Wang Jun,Zheng Hao,Lu Wenhua,Tian Cuihua,Yuan Jian,Chen Baichao,Yuan jiaxin 보안공학연구지원센터 2015 International Journal of Smart Home Vol.9 No.10

As a kind of reactive power compensation equipment with control flexibility and high reliability, magnetically controlled reactor (MCR) is widely used in reactive power compensation, over-voltage limitation and other aspects. However, low responding speed has severely limited the application of MCR. Especially when applied to suppress voltage flicker and arc suppression coil, the slow response of MCR will decrease the stability of control system, even causing the system shock. To improve the response performance of MCR, in this paper, the working principle of MCR has been analyzed, and a fast response structure of MCR has been designed with the novel fast response structure, fast excitation and rapid demagnetization can be achieved. According to the simulation and experiment results, the effectiveness of the proposed structure is verified by limiting the response time in 30ms.

Discovery and validation of PURA as a transcription target of 20(S)-protopanaxadiol: Implications for the treatment of cognitive dysfunction

Feiyan Chen,Wenjing Zhang,Shuyi Xu,Hantao Zhang,Lin Chen,Cuihua Chen,Zhu Zhu,Yunan Zhao 고려인삼학회 2023 Journal of Ginseng Research Vol.47 No.5

Background: 20(S)-protopanaxadiol (PPD), a ginsenoside metabolite, has prominent benefits for thecentral nervous system, especially in improving learning and memory. However, its transcriptionaltargets in brain tissue remain unknown. Methods: In this study, we first used mass spectrometry-based drug affinity responsive target stability(DARTS) to identify the potential proteins of ginsenosides and intersected them with the transcriptionfactor library. Second, the transcription factor PURA was confirmed as a target of PPD by biolayerinterferometry (BLI) and molecular docking. Next, the effect of PPD on the transcriptional levels of targetgenes of PURA in brain tissues was determined by qRT-PCR. Finally, bioinformatics analysis was used toanalyze the potential biological features of these target proteins. Results: The results showed three overlapping transcription factors between the proteomics of DARTSand transcription factor library. BLI analysis further showed that PPD had a higher direct interaction withPURA than parent ginsenosides. Subsequently, BLI kinetic analysis, molecular docking, and mutations inkey amino acids of PURA indicated that PPD specifically bound to PURA. The results of qRT-PCR showedthat PPD could increase the transcription levels of PURA target genes in brain. Finally, bioinformaticsanalysis showed that these target proteins were involved in learning and memory function. Conclusion: The above-mentioned findings indicate that PURA is a transcription target of PPD in brain,and PPD upregulate the transcription levels of target genes related to cognitive dysfunction by bindingPURA, which could provide a chemical and biological basis for the study of treating cognitive impairmentby targeting PURA.

Protein target identifi cation of ginsenosides in skeletal muscle tissues: discovery of natural smallmolecule activators of muscle-type creatine kinase

Feiyan Chen,Kexuan Zhu,Lin Chen,Liufeng Ouyang,Cuihua Chen,Ling Gu,Yucui Jiang,Zhongli Wang,Zixuan Lin,Qiang Zhang,Xiao Shao,Jianguo Dai,Yunan Zhao 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.3

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, themechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides)in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potentialtarget in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides,had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol(PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in thestudy, was selected as a representative to confirm direct binding and its biological importance. Biolayerinterferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPDspecifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by moleculardocking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activityin vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the functionof the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delayingexercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginsengreduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can helpin further developing better CK-MM activators based on the dammarane-type triterpenoid structure.

Target engagement of ginsenosides in mild cognitive impairment using mass spectrometry-based drug affinity responsive target stability

Zhu, Zhu,Li, Ruimei,Qin, Wei,Zhang, Hantao,Cheng, Yao,Chen, Feiyan,Chen, Cuihua,Chen, Lin,Zhao, Yunan The Korean Society of Ginseng 2022 Journal of Ginseng Research Vol.46 No.6

Background: Mild cognitive impairment (MCI) is a transitional condition between normality and dementia. Ginseng is known to have effects on attenuating cognitive deficits in neurogenerative diseases. Ginsenosides are the main bioactive component of ginseng, and their protein targets have not been fully understood. Furthermore, no thorough analysis is reported in ginsenoside-related protein targets in MCI. Methods: The candidate protein targets of ginsenosides in brain tissues were identified by drug affinity responsive target stability (DARTS) coupled with label-free liquid chromatography-mass spectrometry (LC-MS) analysis. Network pharmacology approach was used to collect the therapeutic targets for MCI. Based on the above-mentioned overlapping targets, we built up a proteineprotein interaction (PPI) network in STRING database and conducted gene ontology (GO) enrichment analysis. Finally, we assessed the effects of ginseng total saponins (GTS) and different ginsenosides on mitochondrial function by measuring the activity of the mitochondrial respiratory chain complex and performing molecular docking. Results: We screened 2526 MCI-related protein targets by databases and 349 ginsenoside-related protein targets by DARTS. On the basis of these 81 overlapping genes, enrichment analysis showed the mitochondria played an important role in GTS-mediated MCI pharmacological process. Mitochondrial function analysis showed GTS, protopanaxatriol (PPT), and Rd increased the activities of complex I in a dose-dependent manner. Molecular docking also predicted the docking pockets between PPT or Rd and mitochondrial respiratory chain complex I. Conclusion: This study indicated that ginsenosides might alleviate MCI by targeting respiratory chain complex I and regulating mitochondrial function, supporting ginseng's therapeutic application in cognitive deficits.

Volatile Chemical and Carotenoid Profiles in Watermelons [Citrullus vulgaris (Thunb.) Schrad (Cucurbitaceae)] with Different Flesh Colors

Cuihua Liu,Hongyan Zhang,Zhaoyi Dai,Xi Liu,Yue Liu,Xiuxin Deng,Feng Chen,Juan Xu 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.2

Twelve watermelon [Citrullus vulgaris (Thunb.)Schrad (Cucurbitaceae)] cultivars with different flesh colors were analyzed by HPLC, GC-FID, and GC-MS for their differences in carotenoid, soluble sugar, organic acid,and flavor. Results showed that all-trans violaxanthin, 9-cis-violaxanthin, and luteoxanthin were the main carotenoid esters in watermelons with yellow flesh. However,watermelons with red flesh were rich in all-trans lycopene and their cis-isomers. High concentrations of β-carotene and pro-lycopenes were found in watermelon with orangeyellow flesh. Large variations in the sucrose concentration were observed among the different watermelons. Sucrose and/or fructose were the dominant sugars, while citric acid and malic acid were the main organic acids in watermelon flesh. Limonene was detected in the watermelon flesh of all investigated genotypes. Interestingly, partial correlation analysis of the chemical concentrations revealed 2 significant (p<0.01) positive correlations between β-ionone and β-carotene, and between (E)-geranyl acetone and prolycopenes

Study of H2O adsorption on sulfides surfaces and thermokinetic analysis

Cuihua Zhao,Jian-Hua Chen,Xianhao Long,Jin Guo 한국공업화학회 2014 Journal of Industrial and Engineering Chemistry Vol.20 No.2

Adsorption of water on mineral surfaces was studied using density functional theory andmicrocalorimetry technique. The calculation results show that galena and molybdenite are hydrophobic,while pyrite and sphalerite is hydrophilic. Thermokinetic analysis shows that the heat of adsorption is indecreasing order of pyrite, sphalerite, galena and molybdenite, which is in good agreement with thecalculation results. The adsorption kinetics parameters of hydrophobic galena and molybdenite surfacesare close, while those of hydrophilic pyrite and sphalerite surfaces are very different. The adsorption rateof water on the sphalerite surface is larger than that of water on the pyrite surface.

Constraints on sulfide saturation by crustal contamination in the Shitoukengde Cu-Ni deposit, East Kunlun orogenic belt, northern Qinghai-Tibet Plateau, China

Chiyuan Wang,Zhaowei Zhang,Chengjiang Zhang,Cuihua Chen,Yin Li,Bing Qian 한국지질과학협의회 2021 Geosciences Journal Vol.25 No.3

The Shitoukengde Cu-Ni deposit is located in the eastern part of the East Kunlun Orogenic Belt (EKOB). The overall complex underwent emplacement into the surrounding rock, including the gneiss and marble. The ore-bodies exist in complex I, which is composed of peridotite, pyroxenite, and gabbro. The petrogeochemical characteristics of the rocks show that the m/f values range from 1.93 to 5.97, and the Fe3+/ΣFe ratios of the spinel are < 0.3, indicating that the Shitoukengde complexes have the potential to form a Cu-Ni ore deposit. Based on the evidence of marble and gneiss xenoliths, there are higher CaO and Al2O3 contents in the peridotite and pyroxene, there are Nb and Ta depletions, in the Shitoukengde; these results indicate that the ore-forming parental magma has been obviously contaminated by crustal material during the evolution process and that the materials responsible for the contamination mainly originated from the middle-upper crust. The mafic-ultramafic complex, which is closed to marble, has higher CaO and Al2O3 contents; higher (87Sr/86Sr)i values and lower Ni contents; thus, it is reasonable to infer that the Shitoukengde mafic-ultramafic rocks assimilated some marble, and the contamination of marble without sulfide suppresses the sulfide saturation. The pyroxenite with better mineralization is mainly orthopyroxene, and the increase in the sulfur isotope values in the pyroxenite and peridotite is accompanied by an increase in the Ni content; these results suggest that the SiO2 and S of biotite-plagioclase gneisses, especially the S addition, were important for the Shitoukengde intrusion to reach sulfide saturation. The whole-rock Ni content in the peridotite shows a positive relationship with the FeO content of spinel, indicating that the crystallization of spinel has little effect on sulfide saturation. The capable of crystallizing olivine with Fo content as high as 89.3 mol% and olivine Fo value exhibits a positive relationship with the olivine NiO content, indicating that olivine fractional crystallization did not play a significant role in sulfide saturation. The δ34SV-CDT value of the giant Xiarihamu deposit is much higher than that of Shitoukengde. The degree of crustal S contamination is probably responsible for the difference in the scale and ore grade of the Cu-Ni sulfide deposits in the East Kunlun area. The above indicators could guide the exploration and evaluation of similar deposits in the EKOB.