Ethanol extract from radix of Actinidia chinensis and Evodiamine regulated CCR7 expression in SW480 cells

Objective To identify anti-tumor effects of the ethanol extract from radix of Actinidia chinensis(EERAC) and Evodiamine(Evo) on colon carcinoma SW480 cells,and investigate their regulation of CCR7 expression and the underlying mechanism in colon carcinoma.Methods Radix of Actinidia chinensis was extracted by ethanol.MTT assay and growth curve method were used to detect the inhibition of SW480 cells after being treated with EERAC and Evo.RT-PCT and Western-blot were used to detect the CCR7 expression regulated by EERAC and Evo at the level of protein and mRNA respectively in SW480 cells.Results MTT results showed that EERAC and Evo significantly inhibited the growth of SW480 cells with obvious time-and dose-dependent effect.The 50% inhibitory concentration(IC50) of EERAC was 9.14 mg/mL,while the IC50 of Evo was 2.11 μg/mL.RT-PCR results showed that the expression of CCR7 mRNA in EERAC and Evo group was 29.99% and 18.03% respectively,which was obviously lower than that(56.27%) in control group.Western-blot proved that the expression of CCR7 protein in EERAC and Evo group was 29.03% and 15.28% respectively,which was also significantly reduced when compared to that of 42.48% in control group.Conclusion Ethanol extract from radix of Actinidiae chinensis and Evodiamine could effectively inhibit proliferation and growth of SW480 cells,significantly down-regulate the functional expression of CCR7,and thereby suppress the tumor metastasis.

Combination of Evodiamine with Berberine Reveals a Regulatory Effect on the Phenotypic Transition of Colon Epithelial Cells Induced by CCD-18Co

Objective Transdifferentiation exists between stromal cells or between stromal cells and cancer cells.Evodiamine and berberine are predominant pharmacological components of Zuojin pill,a prescription of Traditional Chinese Medicine,playing crucial functions in remolding of tumor microenvironment.This study aimed to explore the effect of combination of evodiamine with berberine(cBerEvo)on the phenotypic transition of colon epithelial cells induced by tumor-associated fibroblasts,as well as the involved mechanisms.Methods Human normal colon epithelial cell line HCoEpiC cells were treated with the prepared conditioned medium of CCD-18 Co,a human colon myofibroblast line,to induce epithelial-mesenchymal transition.Phase contrast microscope was used to observe the morphological changes.Epithelial-mesenchymal transition markers including E-cadherin,vimentin and alpha-smooth muscle actin(α-SMA)were observed with immunofluorescence microscopy.Migration was assessed by wound healing assay.Western blotting was used to detect the expressions of E-cadherin,vimentin,α-SMA,Snail,ZEB1 and Smads.Results In contrast to the control,the tumor-associated fibroblasts-like CCD-18 Co cells induced downregulation of E-cadherin and up-regulation of vimentin,α-SMA,Snail and ZEB1(P<0.05),and promoted migration of HCoEpiCs(P<0.05),with over expression of Smads including Smad2,p-Smad2,Smad3,p-Smad3 and Smad4(P<0.05),which were abolished by a transforming growth factor-β(TGF-β)receptor inhibitor LY364947 and by cBerEvo in a concentration dependent manner.In addition,cBerEvo-inhibited ratios of p-Smad2/Smad2 and p-Smad3/Smad3 were also dose dependent.Conclusion The above results suggest that cBerEvo can regulate the differentiation of colon epithelial cells induced by CCD-18 Co through suppressing activity of TGF-β/Smads signaling pathway.

Evodiamine derivatives improve cognitive abilities in APP^(swe)/PS1^(ΔE9) transgenic mouse models of Alzheimer's disease

Background: Alzheimer's disease(AD) is a complex neurodegenerative disease. Due to the complexity of its molecular pathogenesis and the interaction of the numerous factors involved, the etiology and pathogenesis of AD have not been fully elucidated. Therefore, effective treatment for AD remains to be developed. Evodiamine, a quinolone alkaloid, has been found to improve learning and memory ability to in the APP^(swe)/PS1^(ΔE9) mouse model of dementia. However, the cytotoxicity and physicochemical properties of evodiamine have limited its use in the treatment of AD.Methods: Evodiamine and its derivatives were effectively synthesized by EDCImediated condensation at room temperature. These target compounds contained 1 thio-and 21 oxo-evodiamine derivatives with different substituted groups. The cytotoxicity of evodiamine and its derivatives and the neuroprotective effects of the evodiamine derivatives against H_2O_2-induced cell loss in SH-SY5 Y cells were investigated using the WST-8 assay. The Morris water-maze test was used to detect the effect of evodiamine and its derivatives on improving learning and memory in APP^(swe)/PS1^(ΔE9) mice.Results: In this study, a series of oxo-and thio-evodiamine derivatives was synthesized. Several derivatives showed lower cytotoxicity and stronger neuroprotective effects than evodiamine and elicited enhanced cognitive improvement, especially in the test of spatial memory in APP^(swe)/PS1^(ΔE9) mice.Conclusion: Our study provides insights for developing novel evodiamine derivatives for chemical intervention and treatment of AD.

A new strategy of enhancing absorptivity and safety for evodiamine: polyamidoamine derivatives modified by polyethylene glycol chain

Background:Traditional Chinese medicine involves complex ingredients and mixtures of ingredients that often exhibit low bioavailability,and excipients are often lacking to increase the absorption-enhancing effects.This study modified the generation 4 polyamidoamine dendrimer with polyethylene glycol of different molecular weights(5000,2000,1000)to form a series of polyamidoamine-co-polyethylene glycol(PAMAM-co-PEG)as a novel class of oral absorption enhancers.Evodiamine,the major alkaloid found in the traditional Chinese medicine Wu Zhu Yu(Fructus Evodiae),was used as a model drug to verify the absorption-enhancing effects and the safety of this alkaloid.Methods:This study utilized the solubility determination method documented in the Pharmacopoeia of the People’s Republic of China(2015 edition)and the D0 values recommended in the US FDA guidelines to comprehensively evaluate the solubility of evodiamine.The permeability of evodiamine was assessed using the apparent permeability coefficient in experiments based on in vitro cell models.Multiple aspects of the biological safety of PAMAM-co-PEG were explored using the MTT assay,LDH assay,and total protein release of the rat intestinal tract.Moreover,the absorption-enhancing effects of PAMAM-co-PEG at different molecular weights on evodiamine were verified via the use of in vitro cell models and in vivo intestinal loop circulation experiments with rats.Results:Evodiamine exhibited low solubility and permeability and was classified into class IV compounds using the biopharmaceutical classification system.PAMAM-co-PEG 2000 demonstrated improvement in the biosafety and absorption-enhancement effect of evodiamine at a specific concentration.This study showed that 0.05%(w/v)of PAMAM-co-PEG 2000 increased the cumulative penetration of evodiamine via cell transport by 1.32 times,and 0.10%(w/v)of PAMAM-co-PEG 2000 increased the area under curve value of evodiamine by 1.31 times.Conclusion:Evodiamine possesses low solubility and permeability and leads to poor oral bioavailability and a certain degree of cytotoxicity.PAMAM-co-PEG 2000 was found to be a potentially safe and efficient oral absorption enhancer.The results of this study might create a foundation for the development of novel excipients suitable for the complex active ingredients of traditional Chinese medicine.

Research progress on the mechanism of evodiamine against gastrointestinal tumor

Evodiamine is an indole quinazoline alkaloid with anti-tumor activity,which may be developed into a drug for the treatment of tumor.It was found that evodiamine could inhibit or even block the signal pathways of Wnt/β-catenin,mTOR,NF--κB,PI3K/AKT,Hippo-YAP and BMP in many kinds of cancer cells,thus interfering with cell division and differentiation,increasing the rate of apoptosis,and down-regulating the expression of various cancer cell markers.Such as B-cell lymphoma 2(Bcl-2),B-cell lymphoma super-large(Bcl-XL),cyclooxygenase 2(COX-2),Survivin,vascular endothelial growth factor(VEGF)and matrix metallopeptidase 9(MMP-9),which have a strong inhibitory effect on asymmetric division,malignant proliferation and angiogenesis of tumor cells.This paper summarizes evodiamine in digestive system tumors by regulating Wnt/β-catenin,mTOR,PI3K/AKT,BMP and other signal pathways,interfering with cell division,differentiation and apoptosis,inhibiting the expression of tumor-related marker genes,and preventing tumor cell migration and invasion,so as to provide a basis for the clinical application and further research of evodiamine in the future.

Antitumor Activity and Theoretical Calculation of Evodiamine and Rutaecarpine

The antitumor activities of two alkaloids, evodiamine（EVO） and rutaecarpine（RUT）, against MCF-7, SMMC-7721 and SW-1353 cells growth in vitro were investigated by MTT assay. The results showed that the anti-tumor effects of two alkaloids were remarkably different. In order to discover the relationship of antitumor activity and structures of the compounds, the dihedral angle, Natural Electron Configuration, frontier molecular orbital profiles（HOMO, LUMO）and bandgaps of these two compounds have been studied based on density functional theory（DFT）by means of DFT-B3LYP/6-31G（d） in Gaussian 03. The calculation results of dihedral angle showed that EVO, due to the existence of methyl group attached to the N（14） atom, have non-planar and twisted structures, which decrease the stability of EVO and increase the activity of EVO. Furthermore, the bandgaps of RUT are lower than that of EVO, indicating RUT has higher stability than EVO, so the activity of EVO is higher than that of RUT. In addition, the negative charge of N14 atom in EVO is lower than that of in RUT, so the positive charge of N（14） atom in EVO is higher than that of in RUT, which suggests that the nucleophile is easier to aggress the N（14） atom in EVO than that in RUT, so the reason of the different antitumor activities of EVO and RUT may be attacked by nucleophile.

Theoretical development and mechanism research of evodiamine's antitumor effect

Evodiamine(EVO),one of the bioactive components of traditional Chinese medicine,has been widely concerned for its anti-tumor properties in recent years.In this review,the anti-tumor mechanism of EVO was reviewed from the aspects of inducing tumor cell apoptosis(internal pathway,external pathway and other pathways),inducing autophagy,inhibiting tumor stem cells,inhibiting cell proliferation and migration,inhibiting peripheral vascular proliferation,and recent research progress of EVO anti-tumor drugs.

Regulatory effects of evodiamine on glucose metabolism-related factors in CT26 colorectal carcinoma-bearing mice

Objective:To investigate the therapeutic effect of evodiamine(EVO)on the expression of hexokinase(HK),lactate dehydrogenase A(LDHA),and pyruvate kinase M(PKM),key enzymes of glycolysis,in the tumor-bearing mice after modeling mouse colon cancer cells(CT26).Methods:A tumor-bearing mouse model was generated by administering axillary injection of CT26 and intraperitoneally injecting different doses of EVO.The therapeutic effects of EVO on CT26 tumor-bearing mice were evaluated by measuring the thymus and spleen indices,tumor volume,tumor suppression rate,and other related indicators in the tumor tissues of mice in each group after the administration of EVO,in addition,histopathological changes in the tumor tissues of the mice in the groups were studied by hematoxylin and eosin staining.The expression levels of HK,LDHA,and PKM in the tumor tissues of each group of mice were measured by performing Western blot to investigate the mechanism of EVO treatment in CT26 tumor-bearing mice.Results:EVO inhibited the growth of tumors in CT26-bearing mice and enhanced their splenic and thymic indices.Western blot results showed that EVO reduced the expression levels of HK,LDHA,and PKM proteins in the tumor tissues of CT26 tumor-bearing mice.Conclusion:EVO has a therapeutic effect on CT26 tumor-bearing mice,and its mechanism of action may be related to the low expression of key enzymes HK,LDHA and PKM of glycolysis in tumor tissues.

Evodiamine activates cellular apoptosis through suppressing PI3K/AKT and activating MAPK in glioma

OBJECTIVE Glioblastoma multiforme(GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis.No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de.veloping new agents to treat GBM is important.This study aimed to evaluate the anti-tumor effect of evodiamine(Evo) on GBM cells,and to determine the underlying mechanisms involved.METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo.diamine(0-10 μmol·L^(-1)) for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS) production was detected using dichlorofluorescein diacetate(DCFH-DA) staining.The changes in mitochondrial mem.brane potential(MMP) were assessed by JC-1 after cells were treated with evodiamine.The expres.sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c,Caspase-3,cleaved Caspase-3,PRAP,and cleaved PARP were measured by Western blot analy.ses.RESULTS According to MTT assay results,Evo significantly inhibited the cell proliferation in a time-and dose-dependent manner.Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species(ROS) production and mitochondrial membrane potential(MMP) disruption.Finally,Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos.phorylation(p38 and JNK,but not ERK) to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas.pase-3,and PARP).CONCLUSION In summary,Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

Simultaneous quantification of evodiamine,rutaecarpine,and dehydroevodiamine in rat cerebrospinal fluid and cerebral nuclei after oral administration by UPLC-MS/MS

Evodiamine,rutaecarpine,and dehydroevodiamine have been demonstrated as the major alkaloids in the fruits of Euodia rutaecarpa,a well-known traditional Chinese medicine with central nervous system activities.To study their cerebrospinal fluid pharmacokinetics and cerebral nuclei distribution,the alkaloids were mixed at the weight ratio of 1:1:1 and orally administered via gavage to the rats at each dose of 15 mg/kg.A quick and reliable ultra-performance liquid chromatographic-tandem mass spectrometry method was developed and applied for the simultaneous analysis of the alkaloids in rat cerebrospinal fluid and cerebral nuclei collected at different time points.Non-compartmental pharmacokinetic profiles were calculated,and the distribution in cerebral nuclei was compared.All the tested compounds were absorbed into rat cerebrospinal fluid and distributed to the brain nuclei quickly.Their distribution in different nuclei varied,as evodiamine mainly in cerebellum and brainstem,rutaecarpine with its maximum in the brainstem,and dehydroevodiamine mostly in the cerebellum and hippocampus.They were eliminated from the brain rapidly without long-time accumulation.In summary,this study revealed the targeting discrepancy of evodiamine,rutaecarpine,and dehydroevodiamine in the brain,and highlighted the possibility for drug candidates in the encephalopathy treatment of the fruits of E.rutaecarpa.

吴茱萸碱调控FOXM1表达促进结肠癌耐药细胞HCT8/5-FU凋亡

目的研究吴茱萸碱(evodiamine,EVO)调控叉头盒蛋白M1(fork-head box protein M1,FOXM1)对结直肠癌耐药细胞HCT8/5-FU增殖和凋亡的影响。方法采用CCK-8法与EdU实验技术,评估EVO对细胞增殖活性的作用效果;通过克隆形成实验,探讨EVO对细胞克隆形成能力的影响;采用流式细胞术对凋亡细胞比例进行量化;蛋白质免疫印迹分析技术检测Bcl-2、Bax、FOXM1、β-catenin、c-MYC、CyclinD1蛋白水平的变化;分子对接探究EVO与FOXM1的相互作用关系;裸鼠移植瘤模型验证EVO在体内对HCT8/5-FU细胞的影响。结果CCK-8实验显示EVO抑制HCT8/5-FU细胞增殖且呈时间和浓度依赖性;EdU实验发现EVO处理组新增殖细胞明显减少;克隆形成实验结果显示EVO明显抑制HCT8/5-FU细胞的克隆形成能力;流式细胞计数发现EVO组细胞的凋亡率明显增高;Western blot显示FOXM1、β-catenin在HCT8/5-FU细胞中明显高表达,EVO能下调细胞中FOXM1、β-catenin、c-MYC、CyclinD1、Bcl-2表达,上调Bax表达;分子对接显示EVO与FOXM1之间具有较强的互作能力;体内实验显示EVO能明显抑制HCT8/5-FU皮下移植瘤的生长且能调控相关蛋白的表达;HE染色发现EVO组的瘤体细胞核固缩和碎裂明显。结论EVO可能通过下调FOXM1蛋白表达抑制Wnt通路激活,从而抑制HCT8/5-FU细胞增殖诱导其凋亡。

Evodiamine inhibits high-fat diet-induced colitis-associated cancer in mice through regulating the gut microbiota

Objective:High-fat diet is one of the main risk factors that disrupt the balance of gut microbiota,which eventually will induce colorectal cancer(CRC).Evodiamine(EVO)is a wildly used multifunctional traditional Chinese medicine extract.In this study,we investigated the role of gut microbiota in high-fat dietpropelled CRC and the potential of EVO for CRC chemoprevention.Methods:Gut microbiota,serum D-lactic acid and endotoxin from 38 patients with colon cancer and 18 healthy subjects were detected by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay(ELISA).In addition,body mass index,phospho-signal transducer and activator of transcription 3(p-STAT3)expression in cancer tissues and paracancerous tissues were detected by immunohistochemistry.A mouse intestinal inflammatory tumor model was established by azomethane/sodium dextran sulfate,followed by treatment with EVO and 5-aminosalicylic acid(ASA).Gut microbiota and inflammatory factors were detected by quantitative polymerase chain reaction,while serum D-lactic acid and endotoxin were detected by ELISA.Furthermore,cell proliferation,cell apoptosis,and interleukin(IL)-6/STAT3/P65 pathway were evaluated by 5-ethynyl-20-deoxyuridine,terminal-deoxynucleotidyl transferase-mediated nick-end labeling,and Western blot assays.Results:In patients with colon cancer,the numbers of Enterococcus faecalis and Escherichia coli were increased,while those of Bifidobacterium,Campylobacter and Lactobacillus were decreased.Serum endotoxin and D-lactic acid levels and p-STAT3 levels were significantly increased.In the mouse model,both EVO and ASA inhibited tumor formation,decreased the proliferation of tumor cells,and induced apoptosis of tumor cells.Compared with the control group,the numbers of E.faecalis and E.coli were decreased,while Bifidobacterium,Campylobacter and Lactobacillus numbers were increased.In the EVO group,serum endotoxin and D-lactic acid levels and inflammatory factors were significantly decreased.Further,the IL6/STAT3/P65 signaling pathway was inhibited in the EVO group.Conclusion:EVO may inhibit the occurrence of colon cancer by regulating gut microbiota and inhibiting intestinal inflammation.The potential mechanism involves inhibition of the IL6/STAT3/P65 signaling pathway,revealing its potential therapeutic significance in clinical applications.

Evodiamine induces extrinsic and intrinsic apoptosis of ovarian cancer cells via the mitogen-activated protein kinase/phosphatidylinositol-3-kinase/protein kinase B signaling pathways

OBJECTIVE: To explore the effects of evodiamine on ovarian cancer cells and the mechanisms underlying such effects.METHODS: Human ovarian cancer cells HO-8910 PM were treated with evodiamine at 0, 1.25,2.5, and 5 μM for 1-4 d. 3-(4,5-Dimethiylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was used to detect the growth inhibition rate of evodiamine-treated HO-8910 PM cells. The cell cycle was observed via propidium iodide(PI) staining. Apoptosis induction was assessed via Annexin V-fluorescein isothiocyanate/propidium iodide(Annexin V-FITC/PI) double staining assay. To verify the mechanism of apoptosis, caspase-dependent apoptotic pathway-related protein was detected by Western blot analysis. The expression levels of mitogen-activated protein kinase(MAPK)and/or phosphatidylinositol-3-kinase(PI3K)/pro-tein kinase B(Akt) pathway-related proteins were also investigated.RESULTS: Evodiamine significantly inhibited the proliferation of HO-8910 PM cells in a dose- and time-dependent manner. Evodiamine induced G2/M arrest with an increase of cyclin B1 level, and promoted cell apoptosis with a decrease of B cell lymphoma/lewkmia-2(Bcl-2) and an increase of Bcl-2-associated X protein(Bax) level. In addition,evodiamine treatment led to the activation of caspase-8, caspase-9, and caspase-3 and the cleavage of poly(ADP-ribose)-polymerase(PARP). Evodiamine targeted the MAPK and/or PI3K/Akt pathways by reducing the expression and activity of PI3 K, Akt, and extracellular signal-regulated kinase mitogen-activated protein kinase(ERK1/2 MAPK)and the activity of p38 MAPK.CONCLUSION: Evodiamine can inhibit the growth of ovarian cancer cells by G2/M arrest and intrinsic and extrinsic apoptosis. In addition, evodiamine-induced PI3K/Akt, ERK1/2 MAPK, and p38 MAPK signaling may be involved in cell death.

Evodiamine-inspired dual inhibitors of histone deacetylase 1(HDAC1) and topoisomerase 2(TOP2) with potent antitumor activity

A great challenge in multi-targetingdrug discovery is to identify drug-like lead compounds with therapeutic advantages over single target inhibitors and drug combinations.Inspired by our previous efforts in designing antitumor evodiamine derivatives,herein selective histone deacetylase 1(HDAC1)and topoisomerase 2(TOP2)dual inhibitors were successfully identified,which showed potent in vitro and in vivo antitumor potency.Particularly,compound 30 a was orally active and possessed excellent in vivo antitumor activity in the HCT116 xenograft model(TGI=75.2%,150 mg/kg,p.o.)without significant toxicity,which was more potent than HDAC inhibitor vorinostat,TOP inhibitor evodiamine and their combination.Taken together,this study highlights the therapeutic advantages of evodiamine-based HDAC1/TOP2 dual inhibitors and provides valuable leads for the development of novel multi-targeting antitumor agents.

Peroxidase-mimicking evodiamine/indocyanine green nanoliposomes for multimodal imaging-guided theranostics for oral squamous cell carcinoma

Here,evodiamine(EVO)and the photosensitizer indocyanine green(ICG)were integrated into a liposomal nanoplatform for noninvasive diagnostic imaging and combinatorial therapy against oral squamous cell carcinoma(OSCC).EVO,as an active component extracted from traditional Chinese medicine,not only functioned as an antitumor chemotherapeutic agent but was also capable of 68Ga-chelation,thus working as a contrast agent for positron emission tomography/computed tomography(PET/CT)imaging.Moreover,EVO could exhibit peroxidase-like catalytic activity,converting endogenous tumor H2O2 into cytotoxic reactive oxygen species(ROS),enabling Chemo catalytic therapy beyond the well-known chemotherapy effect of EVO.As proven by in vitro and in vivo experiments,guided by optical imaging and PET/CT imaging,we show that the theragnostic liposomes have a significant inhibiting effect on in situ tongue tumor through photodynamic therapy combined with chemodynamic chemotherapy.

Evodiamine Inhibits Angiotensin Ⅱ-Induced Rat Cardiomyocyte Hypertrophy

Objective: To investigate the effects of evodiamine(Evo), a component of Evodiaminedia rutaecarpa(Juss.) Benth, on cardiomyocyte hypertrophy induced by angiotensin Ⅱ(Ang Ⅱ) and further explore the potential mechanisms. Methods: Cardiomyocytes from neonatal Sprague Dawley rats were isolated and characterized, and then the cadiomyocyte cultures were randomly divided into control, model(Ang Ⅱ 0.1 μmol/L), and Evo(0.03, 0.3, 3 μmol/L) groups. The cardiomyocyte surface area, protein level, intracellular free calcium([Ca]i) concentration, activity of nitric oxide synthase(NOS) and content of nitric oxide(NO) were measured, respectively. The m RNA expressions of atrial natriuretic factor(ANF), calcineurin(CaN), extracellular signal-regulated kinase-2(ERK-2), and endothelial nitric oxide synthase(e NOS) of cardiomyocytes were analyzed by real-time reverse transcriptionpolymerase chain reaction. The protein expressions of calcineurin catalytic subunit(CnA) and mitogen-activated protein kinase phosphatase-1(MKP-1) were detected by Western blot analysis. Results: Compared with the control group, Ang Ⅱ induced cardiomyocytes hypertrophy, as evidenced by increased cardiomyocyte surface area, protein content, and ANF m RNA expression; increased intracellular free calcium([Ca]i) concentration and expressions of CaN m RNA, CnA protein, and ERK-2 m RNA, but decreased MKP-1 protein expression(P<0.05 or P<0.01). Compared with Ang Ⅱ, Evo(0.3, 3 μmol/L) significantly attenuated Ang Ⅱ-induced cardiomyocyte hypertrophy, decreased the [Ca]i concentration and expressions of CaN m RNA, CnA protein, and ERK-2 m RNA, but increased MKP-1 protein expression(P<0.05 or P<0.01). Most interestingly, Evo increased the NOS activity and NO production, and upregulated the e NOS m RNA expression(P<0.05). Conclusion: Evo significantly attenuated Ang Ⅱ-induced cardiomyocyte hypertrophy, and this effect was partly due to promotion of NO production, reduction of [Ca]i concentration, and inhibition of CaN and ERK-2 signal transduction pathways.

Design, synthesis and biological evaluation of E-ring modified evodiamine derivatives as novel antitumor agents

A series of novel E-ring modified evodiamine derivatives were designed and synthesized as antitumor agents. Their capacity to interfere with the catalytic activity of topoisomerase Ⅰ and Ⅱ was evaluated by the relaxation assay. In vitro antitumor activity results revealed that compound 12 showed good antitumor activity with a broad spectrum. Its binding modes with topoisomerase Ⅰ and Ⅱ were clarified by molecular docking.

吴茱萸碱对哮喘模型大鼠炎症及上皮细胞凋亡的影响及机制

目的 探究吴茱萸碱对哮喘模型大鼠炎症反应及上皮细胞凋亡的影响及潜在机制。方法 将SD大鼠分为对照组、模型组、吴茱萸碱低剂量组(10 mg/kg)、吴茱萸碱高剂量组(20 mg/kg)、地塞米松组(阳性对照,0.5 mg/kg)、表皮细胞生长因子(EGF)组[丝裂原活化蛋白激酶(MAPK)激活剂,10μg]、吴茱萸碱高剂量+EGF组(20 mg/kg吴茱萸碱+10μg EGF),每组10只。除对照组外,其余各组大鼠以10%卵白蛋白(OVA)-氢氧化铝混合液3点注射致敏联合2%OVA雾化液吸入激发的方式建立哮喘模型。检测各组大鼠支气管肺泡灌洗液(BALF)中的炎症细胞(巨噬细胞、淋巴细胞)数,观察大鼠肺组织的病理变化,检测大鼠肺组织中气道上皮细胞凋亡情况、血清炎症因子(肿瘤坏死因子α、白细胞介素6、白细胞介素4)水平、通路相关蛋白[p38 MAPK、磷酸化p38MAPK(p-p38 MAPK)、信号转导及转录激活因子1(STAT1)]及凋亡相关蛋白[B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)]的表达水平。结果 与对照组比较,模型组大鼠肺组织可见支气管黏膜水肿、肺泡间隔增厚和大量炎症细胞浸润,炎症细胞数显著增多;气道上皮细胞凋亡率、炎症因子水平、p-38 MAPK/p-38 MAPK和Bax、STAT1蛋白表达水平均显著升高,Bcl-2蛋白表达水平和Bcl-2/Bax均显著降低(P<0.05)。与模型组比较,吴茱萸碱低、高剂量组和地塞米松组大鼠肺组织病理改变均有不同程度缓解,上述各指标均显著逆转,而EGF组上述指标的变化趋势则相反(P<0.05);EGF能显著减弱高剂量吴茱萸碱对哮喘大鼠炎症反应的改善作用(P<0.05)。结论 吴茱萸碱可减轻哮喘大鼠炎症反应并减少气道上皮细胞凋亡,其作用机制可能与抑制p38 MAPK/STAT1信号通路有关。

静脉注射吴茱萸碱-甘草酸纳米胶束后吴茱萸碱在大鼠体内药代动力学研究

目的 建立高效液相色谱法(HPLC)测定大鼠血浆中的吴茱萸碱,研究大鼠尾静脉注射吴茱萸碱-甘草酸纳米胶束后的药代动力学情况。方法 大鼠尾静脉注射吴茱萸碱-甘草酸纳米胶束和吴茱萸碱溶液后,采用HPLC测定大鼠血浆中吴茱萸碱的含量,绘制血药浓度-时间曲线,以DAS 2.0软件分析处理药代动力学参数。结果 吴茱萸碱-甘草酸纳米胶束和吴茱萸溶液的Cmax分别为(1.61±0.19)和(0.92±0.11)μg·mL^(-1),AUC_(0~t)分别为(4.76±1.41)和(2.07±0.22)mg·L^(-1)·h,CL分别为(0.67±0.21)和(1.38±0.14)L·h^(-1)·kg^(-1),吴茱萸碱-甘草酸纳米胶束的C_(max)、AUC_(0~t)较吴茱萸碱溶液提高了1.75、2.30倍,CL是吴茱萸碱溶液的0.48倍,C_(max)、AUC_(0~t)和CL差异均有统计学意义(P <0.05)。结论 将吴茱萸碱制备为吴茱萸碱-甘草酸纳米胶束能延长吴茱萸碱在体内的作用时间,延缓吴茱萸碱在体内的代谢。

吴茱萸碱对特应性皮炎模型大鼠的治疗作用

目的探讨吴茱萸碱(Evo)调节环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/cAMP反应元件结合蛋白(CREB)信号通路对特应性皮炎(AD)模型大鼠的治疗作用。方法通过多次2,4-二硝基氯苯(DNCB)涂抹建立AD大鼠模型,随机分为AD组、Evo低剂量(Evo-L,5 mg/kg)组、Evo高剂量(Evo-H,10 mg/kg)组、Evo-H+H-89(5 mg/kg)组、地塞米松(0.1 mg/kg)组,并以正常大鼠为对照组,随后对各组大鼠皮损程度进行评分;腹部取血,检测血清中白细胞介素(IL)-4、cAMP、肿瘤坏死因子(TNF)-α水平;收集皮损组织,检测皮损组织病理学变化、肥大细胞数量、组织中PKA/CREB相关蛋白表达以及IL-4、TNF-αmRNA表达。结果与对照组相比,AD组大鼠血清中cAMP水平、皮损组织中p-PKA/PKA、p-CREB/CREB表达降低,皮损程度评分、IL-4、TNF-α水平、表皮厚度、肥大细胞数、皮损组织中IL-4、TNF-αmRNA表达增加(P<0.05);与AD组相比,Evo-L组、Evo-H组、地塞米松组大鼠血清中cAMP水平、皮损组织中p-PKA/PKA、p-CREB/CREB表达增加,皮损程度评分、IL-4、TNF-α水平、表皮厚度、肥大细胞数、皮损组织中IL-4、TNF-αmRNA表达降低(P<0.05);与Evo-H组相比,与Evo-H+H-89组大鼠血清中cAMP水平、皮损组织中p-PKA/PKA、p-CREB/CREB表达降低,皮损程度评分、IL-4、TNF-α水平、表皮厚度、肥大细胞数、皮损组织中IL-4、TNF-αmRNA表达增加(P<0.05)。结论Evo调节cAMP/PKA/CREB信号通路抑制AD模型大鼠的炎性反应及病理损伤。