Effects of oxymatrine on calcium channels and GABA release in mice under neuropathic pain condition

OBJECTIVE To investigate effects of oxymatrine,an alkaloid from Sophora flavescens Ait.,on high-voltage dependent calcium channel and inhibitory neurotransmitter GABA under neuropathic pain condition.METHODS The partial sciatic nerve ligation(PSNL)was executed on C57/BL6 mice to produce neuropathic pain.Oxymatrine(150 mg·kg-1)was administrated intraperitoneally to PSNL mice.Mechanical hindpaw withdral threshold(MWT)was measured under Von-Frey filament stimulation with up-and-down method.In brain tissue,GABA concentration was measured with ELISA.Change of GABAAreceptor protein expression,N-type calcium channel(Cav2.2)and L-type calcium channel(Cav1.3)protein expressions were detected with Western-blot;intracellular calcium concentration was measured in cultured cortical neurons with Fluo-3/AM fluorescent probe.RESULTS Compared to saline,oxymatrine significantly increased ED50 of MWT on PSNL mice(P<0.05).GABA concentration and GABAAreceptor protein level in brain tissue were decreased in PSNL mice,while administration of oxymatrine increased both GABA concentration and GABAA receptor expression.Intracellular calcium concentration was increased in cultured cortical neurons by oxymatrine treatment,but this phenomenon was not seen under calcium-free condition.Protein expression of Cav2.2,but not Cav1.3,was found to be decreased in the brains of PSNL mice and to be restored to a normal level with oxymatrine administration.CONCLUSION Oxymatrine has analgesic effect on PSNL-induced neuropathic pain in mice.This phenominon relates to the increase of GABA release,GABAAreceptor expression,and also the restoration of expression level of Cav2.2 but not Cav1.3 in brain tissues,which suggesting that Ca2+ flow through Cav2.2 calcium channel may be the key point underlying oxymatrine analgesia.

Longan Aril Reverses H2O2 Cytotoxicity in PC12 Cells via RAS/MEK/ERK Signaling Pathway

[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and MDA content,apoptosis assay and relative protein expression of Aβand p-Tau.The neuroprotective mechanism of LA was studied by using metabolomics and network pharmacology,and the expressions of RAS/MEK/ERK signaling pathway-related proteins were detected by western blotting.[Results]LA could improve the cell survival rate and SOD content,and reduce apoptosis and expression of Aβand p-tau.Inhibition of RAS/MEK/ERK signaling pathway is a possible mechanism of LA neuroprotective effects.[Conclusions]LA has a neuroprotective effects in vitro and be likely to inhibit the process of AD by inhibition of RAS/MEK/ERK signalling pathway.

Prediction of active ingredients and potential mechanisms of Alisma decoction against atherosclerosis:A study based on UHPLC-Q-Orbitrap-HRMS and network pharmacology

Background:Atherosclerosis(AS)has been a potentially life-threatening disease worldwide.Alisma decoction(AD)is a famous Chinese formula in treating AS.However,the active components and potential mechanisms remain unknown.Methods:In this study,the active compositions of AD were analyzed by ultrahigh-performance liquid chromatography quadrupole-orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap-HRMS).The putative targets were predicted and interpreted by a series of bio-informative tools such as Venn,STRING,NetworkAnalyzer,ClusterMarker,and DAVID.Finally,the results were validated by molecular docking using AutoDock.Results:The results showed that alisol A,alisol B,23-o-Acetylalisol B,atractylenolideⅢand atractylenolideⅡwere the major compositions in AD.Thirty targets mainly distributed in three gene clusters were obtained after topological and cluster analysis.KEGG and GO analyses showed that genes in cluster 1 mainly participated in adipocytokine signaling pathway and played an important role in lipid metabolism,while genes in cluster 2 had regulatory actions via cAMP signaling pathway and may protect vascular function from AS,and genes in cluster 3 were closely related to inflammation.Furthermore,RXRA,AGT and CXCL8,the central gene of cluster 1,2 and 3,were verified to be strong binding with some major compositions in AD.Conclusion:The possible mechanisms of AD in the treatment of AS may be closely correlated with the regulation of lipid metabolism,vascular physiology and inflammation.Our study provided a new insight into the anti-AS effect of AD,but more pharmacological experiments should be performed for verification in the future.

New bisabolane-type phenolic sesquiterpenoids from the marine sponge Plakortis simplex

Six new bisabolane-type phenolic sesquiterpenoids,including plakordiols A-D(1-4),(7 R,10 R)-hydroxycurcudiol(5)and(7 R,10 S)-hydroxycurcudiol(6)were isolated from the marine sponge Plakortis simplex collected from the South China Sea.Their structures were determined based on extensive analysis of spectroscopic data.Their configurations were assigned by coupling constant analysis,NOESY correlations,and the modified Mosher’s method.Furthermore,their cytotoxic and antibacterial activities were evaluated.

Analyses and identification of the chemical constituents of the Uighur formulation Baixuan Xiatare using three-stage infrared spectroscopy

OBJECTIVE:To identify,rapidly and accurately,the chemical composition of the traditional Uighur formulation Baixuan Xiatare(BXXTR-FU).METHODS:We investigated if application of three-stage infrared(IR)spectroscopy enabled identification of the main chemical constituents(and their origins)in BXXTR-FU.RESULTS:The characteristic peaks of herbal material(s)and BXXTR-FU were assigned.In Fourier transform-IR(FT-IR)spectroscopy of BXXTR-FU,peaks at1616 and 1605 cm-1 of BXXTR-FU were considered to denote anthraquinones and their derivatives;1066 cm-1 was regarded as the characteristic absorption peak of resin glycosides.In second-derivative IR(SD-IR)spectroscopy,the main carbonyl types of BXXTR-FU in the range 1743-1636 cm-1 were assigned:1651 cm-1 belonged to the carbonyl stretching vibrations of flavonoids and chromones;1717 cm-1 belonged to tannins;1699 cm-1 belonged to carboxylic acids;1636 cm-1 belonged to anthraquinones and their derivatives.SD-IR spectroscopy further confirmed that the characteristicabsorption peaks at 1636,1618 and 1603 cm-1 could be used as markers that BXXTR-FU contained anthraquinones and their derivatives.Synchronous2 D-IR correlation spectra of chemical groups further confirmed the results of FT-IR and SD-IR spectroscopy.CONCLUSION:Our study strongly supported the necessity and importance of three-stage IR spectroscopy owing to its rapid and accurate identification of herbal formulations.

Shenmai injection enhances cisplatin-induced apoptosis through regulation of Mfn2-dependent mitochondrial dynamics in lung adenocarcinoma A549/DDP cells

Aim:Chemoresistance is the biggest obstacle in cancer treatment.Our previous study demonstrated that Shenmai injection(SMI),a Chinese herbal medicine,enhanced the antitumor effect of cisplatin via glucose metabolism reprogramming.This study aimed to further determine whether the SMI sensitizes the non-small cell lung cancer(NSCLC)cells to cisplatin through regulation mitochondrial dynamics.Methods:The Kaplan-Meier Plotter database was used to investigate the relationship between mRNA expression of mitofusin-2(Mfn2)and the survival analysis of NSCLC patients.The protein expression of Mfn2 in a lung adenocarcinoma tissue chip was detected by immunohistochemistry staining.The expression of Mfn2 and ATAD3A were compared between cisplatin-sensitive A549 and cisplatin-resistant A549/DDP cells.Additionally,A549/DDP cells were co-treated with cisplatin and SMI to detect mitochondrial morphology by fluorescent staining,apoptosis-related protein expression with Western blotting,and mitochondrial membrane potential(ΔΨm)with flow cytometry analysis.Results:The mean survival time of the Mfn2^(low) group was significantly lower than that of the Mfn2^(high) group by Kaplan-Meier Plotter database analysis,and the Mfn2 protein expression level was lower in cancer tissues than in adjacent tissues.The combination of SMI and cisplatin induced dynamic changes in A549/DDP cells,with increased mitochondrial fusion followed by upregulation of Mfn2 and downregulation of ATAD3A and reduced mitochondrial mass and ΔΨm.Moreover,SMI significantly enhanced cisplatin-induced A549/DDP apoptosis,upregulated Bax and the active subunit of caspase-3,and downregulated Bcl-2 expression,as shown via Hoechst staining and flow cytometry analysis.Conclusion:Our findings suggest SMI enhances cisplatin-induced apoptosis through regulation of Mfn2-dependent mitochondrial dynamics in cisplatin-resistant lung adenocarcinoma cells.