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1.
Pak J Pharm Sci ; 32(3): 1103-1105, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31278726

RESUMO

Invasive fungal infections caused by Candida albicans constitute a prevalent worldwide health problem. Due to limited antifungal agents available, more efforts have been made towards searching the novel anti-candida drugs with low cytotoxicity. The present study was aimed to investigate the antifungal activities of baicalin and/or sodium bicarbonate (SB) against 29 C. albicans isolates including 27 clinical ones. By using broth microdilution method and checkerboard assay, it was observed that the minimum inhibitory concentrations (MICs) of baicalin and SB alone were > 2048 µg/mL, and those of baicalin and SB in combination decreased 16-32 folds with fractional inhibitory concentration index (FICI) in a range of 0.094-0.375. The results presented the strong synergism between SB and baicalin in 27 clinical C. albicans isolates and provided an alternative choice against C. albicans.

2.
Phytomedicine ; 62: 152948, 2019 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-31129431

RESUMO

BACKGROUND: Huangqi decoction (HQD), a classic traditional herbal medicine, has been used for liver fibrosis, but its effect on intrahepatic chronic cholestatic liver injury remains unknown. PURPOSE: In the present study, we investigated the hepatoprotective effect of HQD and the underlying molecular mechanisms in 3, 5-diethoxycarbonyl-1, 4-dihydroxychollidine (DDC)-induced chronic cholestatic mice. METHODS: The DDC-induced cholestatic mice were administrated HQD for 4 or 8 weeks. Serum biochemistry and morphology were investigated. The serum and liver bile acid (BA) levels were detected by ultra performance liquid chromatography-tandem mass spectrometry. The liver expression of BA metabolizing enzymes and transporters, and inflammatory and fibrotic markers was measured by real-time polymerase chain reaction, western blotting, and immunohistochemistry. RESULTS: HQD treatment for 4 or 8 weeks ameliorated DDC-induced liver injury by improving impaired hepatic function and tissue damage. HQD treatment for 8 weeks further decreased the liver expression of cytokeratin 19, tumor growth factor (TGF)-ß, collagen I, and α-smooth muscle actin, and ameliorated ductular reaction and liver fibrosis. HQD markedly decreased the accumulation of serum and liver BA. The expression of BA-metabolizing enzymes, cytochrome P450 2b10 and UDP glucuronosyltransferase 1 A1, and multidrug resistance-associated protein 2, Mrp3, and Mrp4 involved in BA homeostasis was increased by 4 weeks of HQD treatment. The expression of BA uptake transporter Na+-taurocholate cotransporting polypeptide was decreased and that of Mrp4 was increased after 8 weeks of HQD treatment. Nuclear factor-E2-related factor-2 (Nrf2) was remarkably induced by HQD treatment. Additionally, HQD treatment for 8 weeks decreased the liver expression of inflammatory factors, interleukin (IL)-6, IL-1ß, tumor necrosis factor-α, monocyte chemoattractant protein-1, and intracellular adhesion molecule-1. HQD suppressed the nuclear factor (NF)-κB pathway. CONCLUSION: HQD protected mice against chronic cholestatic liver injury and biliary fibrosis, which may be associated with the induction of the Nrf2 pathway and inhibition of the NF-κB pathway, ameliorating BA-stimulated inflammation.

3.
Zhongguo Zhong Yao Za Zhi ; 44(2): 350-356, 2019 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-30989957

RESUMO

This study aimed to investigate the effect of butyl alcohol extract of Baitouweng Decoction( BAEB) on Candida albicans biofilms based on pH signal pathway. The morphology of biofilms of the pH mutants was observed by scanning electron microscope. The biofilm thickness of the pH mutants was measured by CLSM. The biofilm activity of the pH mutants was analyzed by microplate reader.The biofilm damage of the pH mutants was detected by flow cytometry. The expression of pH mutant biofilm-related genes was detected by qRT-PCR. The results showed that the deletion of PHR1 gene resulted in the defect of biofilm,but there were more substrates for PHR1 complementation. BAEB had no significant effect on the two strains. RIM101 gene deletion or complementation did not cause significant structural damage,but after BAEB treatment,the biofilms of both strains were significantly inhibited. For the biofilm thickness,PHR1 deletion or complementation caused the thickness to decrease,after BAEB treatment,the thickness of the two strains did not change significantly. However,RIM101 gene deletion or complementation had little effect on the thickness,and the thickness of the two strains became thinner after adding BAEB. For biofilm activity,PHR1 deletion or complementation and RIM101 deletion resulted in decreased activity,RIM101 complementation did not change significantly; BAEB significantly inhibited biofilm activity of PHR1 deletion,PHR1 complemetation,RIM101 deletion and RIM101 complemetation strains. For the biofilm damage,PHR1 gene deletion or complementation,RIM101 gene deletion or complementation all showed different degrees of damage; after adding BAEB,the damage rate of PHR1 deletion or complementation was not significantly different,but the damage rate of RIM101 deletion or complementation was significantly increased. Except to the up-regulation of HSP90 gene expression,ALS3,SUN41,HWP1,UME6 and PGA10 genes of PHR1 deletion,PHR1 complementation,RIM101 deletion,and RIM101 complementation strains showed a downward expression trend. In a word,this study showed that mutations in PHR1 and RIM101 genes in the pH signaling pathway could enhance the sensitivity of the strains to the antifungal drug BAEB,thus inhibiting the biofilm formation and related genes expression in C. albicans.


Assuntos
Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Extratos Vegetais/farmacologia , Transdução de Sinais , 1-Butanol , Proteínas Fúngicas , Regulação Fúngica da Expressão Gênica , Concentração de Íons de Hidrogênio
4.
Biomed Pharmacother ; 112: 108701, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30818137

RESUMO

Natural bear bile has been used for liver disease in East Asia for thousands of years. However, its use has restrictions. In the current study, the therapeutic effects and potential mechanisms of cultured bear bile powder (CBBP) against hepatic fibrosis were evaluated in a dimethylnitrosamine (DMN)-induced rat model. CBBP treatment significantly improved DMN-induced hepatic necrosis and inflammatory infiltration. Additionally, CBBP remarkably alleviated the increased hepatic collagen content and expression of alpha-smooth muscle actin. Serum metabolomics revealed that 14 serum metabolites, including docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were decreased in DMN-treated rats, which was reversed by CBBP. Pathway analyses revealed that the main metabolic pathways affected by CBBP were related to fatty acid biosynthesis and metabolism, and biosynthesis of unsaturated fatty acids. EPA and DHA are ligands of peroxisome proliferator activated receptors (PPARs). CBBP treatment significantly stimulated liver mRNA and protein expression of PPARα and PPARγ. CBBP also markedly increased liver expression of PPARα target genes, which are involved in fatty acid ß-oxidation, and down-regulated IL-6, a downstream inflammatory gene of PPARγ. In conclusion, CBBP has the potential to attenuate liver fibrosis and its mechanism involves the promotion of the liver expression of PPARα and PPARγ. Our results may help in the development of a novel substitute for bear bile and therapeutic strategies for fibrotic liver diseases.


Assuntos
Bile/metabolismo , Citoproteção/efeitos dos fármacos , Dimetilnitrosamina/toxicidade , Cirrose Hepática Experimental/induzido quimicamente , Cirrose Hepática Experimental/metabolismo , Animais , Bile/química , Citoproteção/fisiologia , Relação Dose-Resposta a Droga , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/metabolismo , Cirrose Hepática/prevenção & controle , Cirrose Hepática Experimental/prevenção & controle , Distribuição Aleatória , Ratos , Ratos Wistar , Ursidae
5.
Zhongguo Zhong Yao Za Zhi ; 44(1): 125-130, 2019 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-30868823

RESUMO

The aim of this paper was to investigate the inhibitory effect of extract of Coptidis Rhizoma(ECR) on invasion of Candida albicans hyphae in vitro.XTT reduction method was used to evaluate the metabolic activity of C.albicans.The colony edge growth of C.albicans was observed by solid medium.The growth of C.albicans hyphae was determined on semi-solid medium.The morphology and viability changes of C.albicans hyphae were assessed by scanning electron microscope and fluorescence microscope.qRT-PCR method was used to detect the ALS3 and SSA1 expression of C.albicans invasin genes.The results showed that the metabolic viability by XTT method detected that the activity of C.albicans was gradually decreased under the intervention of 64,128 and 256 mg·L-1 of ECR respectively.128,256 mg·L-1 of ECR significantly inhibited colony folds and wrinkles on solid medium and the hyphal invasion in semi-solid medium.Scanning electron microscopy and fluorescence microscopy showed that 128,256 mg·L-1 of ECR could inhibit the formation of C.albicans hyphae.qRT-PCR results showed that the expression of invasin gene ALS3 and SSA1 was down-regulated,and especially 256 mg·L-1 of ECR could down-regulate the two genes expression by 4.8,1.68 times respectively.This study showed that ECR can affect the invasiveness of C.albicans by inhibiting the growth of hyphae and the expression of invasin.


Assuntos
Candida albicans/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Proteínas Fúngicas/genética , Adenosina Trifosfatases/genética , Regulação Fúngica da Expressão Gênica , Proteínas de Choque Térmico HSP70/genética , Hifas/efeitos dos fármacos , Hifas/ultraestrutura , Microscopia Eletrônica de Varredura
6.
Zhongguo Zhong Yao Za Zhi ; 43(14): 2979-2984, 2018 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-30111058

RESUMO

To investigate the mechanism of n-butanol extract of Pulsatilla decoction (BAEB) against murine ulcerative colitis (UC) model induced by DSS combined with Candida albicans (CA) colonization, mice were randomly divided into normal control group, DSS group, DSS+CA group, BAEB high, medium and low dose group, and positive drug Mesalazine group. The general condition of mice was observed, fungal loads of murine intestinal contents were detected by plate method, colonic pathological change of mice was examined by HE staining. ASCA in serum and IL-6, IL-8, IL-1ß, HBD-2, HBD-3 in colonic mucosa were detected by ELISA. The results showed that, compared with DSS group, the general condition and ASCA in serum had no obvious change for DSS+CA group, but the fungal loads in intestinal contents, the colonic pathological damage, and the levels of IL-6, IL-8, IL-1ß, HBD-2, HBD-3 in colonic mucosa were greater than that of DSS group. High dose of BAEB group and Mesalazine group could improve the colonic pathology, decrease IL-6, IL-8, IL-1ß, HBD-2, HBD-3 expression level. In conclusion, BAEB could effectively improve the UC symptoms in mice induced by DSS combined with CA colonization, and inhibit the inflammatory factors such as IL-6, imply that BAEB is of important value for the treatment of intestinal fungal-related colitis.

7.
Biomed Chromatogr ; 32(5): e4178, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29243282

RESUMO

A rapid, sensitive and accurate UPLC-MS/MS method was developed for the simultaneous quantification of components of Huangqi decoction (HQD), such as calycosin-7-O-ß-d-glucoside, calycosin-glucuronide, liquiritin, formononetin-glucuronide, isoliquiritin, liquiritigenin, ononin, calycosin, isoliquiritigenin, formononetin, glycyrrhizic acid, astragaloside IV, cycloastragenol, and glycyrrhetinic acid, in rat plasma. After plasma samples were extracted by protein precipitation, chromatographic separation was performed with a C18 column, using a gradient of methanol and 0.05% acetic acid containing 4mm ammonium acetate as the mobile phase. Multiple reaction monitoring scanning was performed to quantify the analytes, and the electrospray ion source polarity was switched between positive and negative modes in a single run of 10 min. Method validation showed that specificity, linearity, accuracy, precision, extraction recovery, matrix effect and stability for 14 components met the requirements for their quantitation in biological samples. The established method was successfully applied to the pharmacokinetic study of multiple components in rats after intragastric administration of HQD. The results clarified the pharmacokinetic characteristics of multiple components found in HQD. This research provides useful information for understanding the relation between the chemical components of HQD and their therapeutic effects.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/farmacocinética , Espectrometria de Massas em Tandem/métodos , Administração Oral , Animais , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Feminino , Flavonoides/sangue , Flavonoides/química , Flavonoides/farmacocinética , Glucosídeos/sangue , Glucosídeos/química , Glucosídeos/farmacocinética , Glicosídeos/sangue , Glicosídeos/química , Glicosídeos/farmacocinética , Modelos Lineares , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Terpenos/sangue , Terpenos/química , Terpenos/farmacocinética
8.
Oncotarget ; 8(57): 97137-97152, 2017 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-29228599

RESUMO

This study explored the effects of chicken bile powder (CBP), a 2000-year-old Chinese medicine, on α-naphthyl isothiocyanate (ANIT)-induced intrahepatic cholestasis in mice. CBP treatment for 14 days significantly ameliorated ANIT-induced changes in serum alanine aminotransferase, aspartate aminotransferase, bile acids, bilirubin, γ-glutamyl transpeptidase, alkaline phosphatase, and liver tissue morphology. Serum metabolomics showed changes in 24 metabolites in ANIT-exposed mice; 16 of these metabolites were reversed by CBP treatment via two main pathways (bile acid biosynthesis and arachidonic acid metabolism). Additionally, CBP administration markedly increased fecal and biliary bile acid excretion, and reduced total and hydrophobic bile acid levels in the livers of cholestatic mice. Moreover, CBP increased liver expression of bile acid efflux transporters and metabolic enzymes. It also attenuated ANIT-induced increases in hepatic nuclear factor-κB-mediated inflammatory signaling, and increased liver expression of the nuclear farnesoid X receptor (FXR) in cholestatic mice. CBP also activated FXR in vitro in HEK293T cells expressing mouse Na+-taurocholate cotransporting polypeptide. It did not ameliorate the ANIT-induced liver injuries in FXR-knockout mice. These results suggested that CBP provided protection from cholestatic liver injury by restoring bile acid homeostasis and reducing inflammation in a FXR-dependent manner.

9.
Zhongguo Zhong Yao Za Zhi ; 42(21): 4201-4206, 2017 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-29271161

RESUMO

To investigate the inhibitory effect and mechanism of chloroform extracts from Longdan Xiegan decoction(CELX) against hydrolytic enzymes activity of Candida albicans isolated from vulvovaginal candidiasis(VVC) patients. Secreted aspartyl proteinase(Sap), phospholipase(PL) and lipase(Lip) positive strains were identified from 15 strains of C. albicans with milk culture medium, egg yolk culture medium and tween-80 medium, respectively. Then, the activities of Sap, PL, and Lip were detected in the above media. qRT-PCR was used to detect the changes in gene expressions of aspartic protease(SAP1-7,10), phospholipase B(PLB1-2) and lipase(LIP3-6). Secreted aspartyl proteinase and phospholipase of 15 VVC clinical strains were positive, and lipase of 11 strains were positive. Compared with the blank control group, the drug CELX-containing medium(milk medium, egg yolk culture medium, tween-80 medium) experiment showed that the sedimentation of colonies decreased gradually in each culture medium with the increase of CELX dose. When the concentration of CELX was 256 mg•L⁻¹, the colony almost disappeared, which indicated the enzyme activity was significantly weakened. The results of qRT-PCR showed that SAP1, SAP2, SAP3, SAP4, SAP7, SAP9 and SAP10 were down-regulated by 62%, 55%, 62%, 84%, 61%, 51%, 68%, respectively, except for SAP5 and SAP6; and PLB1, LIP3, LIP4, LIP6 were down-regulated by 67%, 51%, 54%, 55%, respectively. The findings suggested that CELX may inhibit the activities of Sap, PL, and Lip, which are important virulence factors of C. albicans.

10.
Zhongguo Zhong Yao Za Zhi ; 42(18): 3572-3577, 2017 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-29218944

RESUMO

To explore the activity of essential oil extracted from Artemisia argyi (AAEO) in inducing the apoptosis of Candida albicans SC5314. The effect of AAEO on reactive oxygen species(ROS) and mitochondria membrane potential(MMP) of C. albicans SC5314 was detected by flow cytometry. Phosphatidylserine externalization was observed under fluorescence microscopic with Annexin-V/PI staining at the early stage of apoptosis in C. albicans. Metacaspase activity was observed under fluorescence microscopic with FITC-VAD-FMK staining at the early stage of apoptosis in C. albicans. C. albicans morphology was observed by DAPI nuclear staining and fluorescence microscopy. After intervention with 0.5 mL•L⁻¹ AAEO, apoptosis of C. albicans significantly increased, metacaspase activity increased, nuclear pyknosis and fragmentation, and intracellular ROS were significantly increased, and mitochondrial membrane potential decreased significantly. The certain concentrations of AAEO could induce the apoptosis of C. albicans.

11.
Zhongguo Zhong Yao Za Zhi ; 42(16): 3182-3190, 2017 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-29171239

RESUMO

To study the inhibitory effect of butyl alcohol extract of Baitouweng decoction(BAEB) on Candida albicans cell membrane. The effects of BAEB on the activity of C. albicans were observed by Spot assay. The changes of intracellular osmotic pressure of C. albicans after BAEB intervention were detected by microtiter plate reader. The effect of BAEB on cell membrane permeability of C. albicans were observed by fluorescence microscopy. The content of ergosterol in C. albicans cell membrane was detected by high performance liquid chromatography, and the expression of ergosterol biosynthesis related genes in cell membrane was detected by qRT-PCR. The results showed that the activity of C. albicans was significantly decreased in 256, 512 and 1 024 mg•L⁻¹ BAEB group. The intracellular glycerol content of C. albicans was significantly increased in 512 and 1 024 mg•L⁻¹ BAEB group(P<0.05). The gene HOG1 associated with intracellular osmotic pressure of C. albicans was down-regulated by 9.1, 9.3 and 5.5 times, respectively. C. albicans with red fluorescent were increased significantly in 512 and 1 024 mg•L⁻¹ BAEB group. The peak area of ergosterol in the 1 024 mg•L⁻¹ BAEB group was 35.884 95, with a significant difference(P<0.05); ERG1, ERG2, ERG3, ERG4, ERG5, ERG6, ERG10, ERG11, ERG13, ERG24, ERG25, ERG251, ERG26 and UPC2 were down-regulated by 6.58, 4.89, 4.15, 9.24,3.41, 9.84, 3.08, 7.50, 5.53, 5.90, 2.45, 3.25,1.98 and 10.07 times respectively in 1 024 mg•L⁻¹ BAEB group. The study indicated that BAEB could inhibit ergosterol and its biosynthesis related genes expression in the cell membrane and inhibit the activity of C. albicans.

12.
Biomed Pharmacother ; 93: 709-712, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28700974

RESUMO

BACKGROUND: Candida tropicalis was one of the most common non-albicans Candida (NACA) species creating relatively high morbidity and mortality. We previously demonstrated that the antifungal effect of BBR in combination with FLC might be attributed to reactive oxygen species production, ergosterol content and efflux pump levels in a clinical C. tropicalis isolate. METHODS: we employed thirteen clinical C. tropicalis isolates as well as four standard reference strains to evaluate the antifungal activity of BBR in combination with FLC under planktonic and biofilm states by analyzing the mRNA expressions of ERG11, CDR1 and MDR1 by qRT-PCR. RESULTS: The susceptibility tests showed that BBR in combination with FLC produced a FICI range of 0.13-1 under planktonic state and 0.25-1 under biofilm state in the seventeen strains. The mRNA expressions of ERG11, CDR1 and MDR1 were upregulated 1.43-2.10 and 1.47-4.42 fold in the case of BBR/FLC used alone compared with the control. The combined usage of BBR plus FLC resulted in a decrease of 2.84- (p<0.01) and 2.39-fold (p<0.05) in the expression of ERG11 mRNA, 2.98- (p<0.05) and 3.06-fold (p<0.05) in the expression of CDR1 mRNA, 1.41- and 4.33-fold in the expression of MDR1 mRNA compared with the BBR/FLC used alone. CONCLUSION: The synergism of BBR with FLC might be relevant with the inhibitions on the mRNA expressions of ERG11 and efflux pumps.


Assuntos
Antifúngicos/farmacologia , Berberina/farmacologia , Candida tropicalis/efeitos dos fármacos , Farmacorresistência Fúngica/efeitos dos fármacos , Fluconazol/farmacologia , Biofilmes/efeitos dos fármacos , Candidíase/tratamento farmacológico , Candidíase/metabolismo , Ergosterol/metabolismo , Proteínas Fúngicas/metabolismo , Humanos , Proteínas de Membrana Transportadoras , Testes de Sensibilidade Microbiana/métodos , RNA Mensageiro/metabolismo
13.
Pharm Biol ; 55(1): 355-359, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27931143

RESUMO

CONTEXT: Fluconazole resistance is an intractable problem of treating Candida albicans, calling for more antifungal agents to enhance the activity of fluconazole. OBJECTIVE: This work investigates the anti-C. albicans activities of sodium houttuyfonate (SH) and/or fluconazole and the associated mechanism. MATERIALS AND METHODS: The minimum inhibitory concentrations (MICs) of SH and fluconazole both ranging from 0.5 to 1024 µg/mL were determined by broth microdilution method in 19 C. albicans isolates, and their fractional inhibitory concentration index (FICI) was evaluated by checkerboard assay. After MICSH and/or MICfluconazole treatments, the expressions of IFD6, PHR1, ZAP1, ADH5, BGL2, XOG1 and FKS1 were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in C. albicans 1601. RESULTS AND CONCLUSION: The MICs of SH alone ranged from 32 to 256 µg/mL and decreased 2-16-fold in combination. SH showed strong synergism with fluconazole with FICI <0.13-0.5. In C. albicans 1601, we observed that (i) the expression of the seven genes increased notably in a range between 3.71- and 12.63-fold (p < 0.05) when SH was used alone, (ii) the combined use of SH and fluconazole slightly inhibited the expression of IFD6 and PHR1 by 1.23- and 1.35-fold (p > 0.05), but promoted evidently the expression of ZAP1, ADH5, XOG1 and FKS1 by 1.98-, 3.56-, 4.10- and 2.86-fold (p < 0.05). The results suggested SH to be a potential synergist to enhance the antifungal activity of fluconazole in C. albicans resistant isolates, and the underlying mechanism may be associated with ß-1,3-glucan synthesis and transportation.


Assuntos
Alcanos/farmacologia , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Farmacorresistência Fúngica/efeitos dos fármacos , Fluconazol/farmacologia , Sulfitos/farmacologia , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/genética , Candida albicans/genética , Candida albicans/crescimento & desenvolvimento , Candida albicans/metabolismo , Metabolismo dos Carboidratos/efeitos dos fármacos , Metabolismo dos Carboidratos/genética , Farmacorresistência Fúngica/genética , Sinergismo Farmacológico , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Reação em Cadeia da Polimerase Via Transcriptase Reversa , beta-Glucanas/metabolismo
14.
Front Pharmacol ; 8: 938, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29311939

RESUMO

Intrahepatic cholestasis is a serious symptom of liver disorders with limited therapies. In this study, we investigated the efficacy of Huangqi decoction (HQD), a two-herb classic traditional Chinese medicine (TCM), in the treatment of alpha-naphthylisothiocyanate (ANIT)-induced intrahepatic cholestasis in mice. HQD treatment ameliorated impaired hepatic function and tissue damage. A metabolomics study revealed that the endogenous metabolites significantly affected by HQD were related to bile acid (BA) biosynthesis and glutathione metabolism pathways. HQD treatment decreased the intrahepatic accumulation of cytotoxic BAs, normalized serum BA levels, and increased biliary and urinary BA excretion. Additionally, HQD restored the hepatic glutathione content and suppressed reactive oxygen species (ROS) in cholestatic mice. Protein and gene analysis revealed that HQD increased the expression of the hepatic metabolizing enzymes cytochrome P450 (CYP) 2B10 and UDP glucuronosyltransferase family 1 member A1 (UGT1A1), as well as multidrug resistance-associated protein 2 (Mrp2), Mrp3, and Mrp4, which play crucial roles in BA homeostasis. Further, HQD increased the protein expression of glutamate-cysteine ligase, which is involved in the synthesis of glutathione. Importantly, HQD increased the nuclear expression of nuclear factor-E2-related factor-2 (Nrf2). In conclusion, HQD protects against intrahepatic cholestasis by reversing the disordered homeostasis of BAs and glutathione.

15.
Front Microbiol ; 7: 1516, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27721812

RESUMO

Candida tropicalis has emerged as an important pathogenic fungus in nosocomial infections due to its recalcitrant resistance to conventional antifungal agents, especially to fluconazole (FLC). Berberine (BBR) is a bioactive herbal-originated alkaloids and has been reported to possess antifungal functions against C. albicans. In this paper, we tried to figure out the antifungal mechanisms of BBR and/or FLC in a clinical C. tropicalis isolate 2006. In the microdilution test, the minimum inhibitory concentration (MIC) of BBR was found 16 µg/mL with fractional inhibitory concentration index (FICI) 0.13 in C. tropicalis 2006. The synergism of BBR and FLC was also confirmed microscopically. After the treatments of BBR and/or FLC, the studies revealed that (i) FLC facilitated BBR to increase reactive oxygen species (ROS), (ii) FLC enhanced the intranuclear accumulation of BBR, (iii) BBR decreased the extracellular rhodamine 123 (Rh123) via inhibiting efflux transporters, (iv) FLC assisted BBR to reduce ergosterol content, and (v) BBR in combined with FLC largely downregulated the expressions of Candida drug resistance 1 (CDR1) and CDR2 but impact slightly multidrug resistance 1 (MDR1), and upregulate the expression of ergosterol 11 (ERG11). These results suggested that BBR could become a potent antifungal drug to strengthen FLC efficacy in FLC-resistant C. tropicalis via ROS increase, intracellular BBR accumulation, ergosterol decrease and efflux inhibition.

16.
J Exp Zool A Ecol Genet Physiol ; 325(7): 453-66, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27455909

RESUMO

Gonadotropin-releasing hormone (GnRH) plays a vital role in the regulation of reproduction through interaction with a specific receptor (the GnRH receptor). In this study, the GnRH receptor gene from the cuttlefish Sepiella japonica (SjGnRHR) was identified and characterized. The cloned full-length SjGnRHR cDNA was 1,468 bp long and contained a 1,029 bp open reading frame encoding 342 amino acid residues, 8 bp of 5' untranslated regions (UTR), and 431 bp of 3' UTR. The putative protein was predicted to have a molecular weight of 38.75 kDa and an isoelectric point of 9.47. In addition, this protein was identified as belonging to the rhodopsin-type (class A) G protein-coupled receptor family. The predicted amino acid sequence contained two N-linked glycosylation sites and 18 phosphorylation sites. Multiple sequence alignment, phylogenetic tree analysis, and three-dimensional structure modeling were conducted to clarify SjGnRHR bioinformatics characteristics. In vitro SjGnRHR expression was carried out using HEK293 cells and the pEGFP-N1 plasmid, to verify the transmembrane properties of this protein. The interaction between the S. japonica GnRH receptor and its ligand was clarified using internalization analysis. SjGnRHR transcriptional quantification confirmed the wide distribution of SjGnRHR in various S. japonica mature tissues. In addition, the transcriptional profile of SjGnRHR in the female brain and ovary during gonadal development was analyzed. Results indicate that GnRHR may be involved in diverse S. japonica physiological functions, especially in the control of reproduction.


Assuntos
Decapodiformes/metabolismo , Expressão Gênica , Receptores LHRH/química , Sequência de Aminoácidos , Animais , Clonagem Molecular , Decapodiformes/fisiologia , Feminino , Humanos , Masculino , Modelos Moleculares , Especificidade de Órgãos , Filogenia , Receptores LHRH/genética , Receptores LHRH/metabolismo , Receptores LHRH/fisiologia , Reprodução , Alinhamento de Sequência
17.
J Exp Zool A Ecol Genet Physiol ; 325(3): 181-93, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27076436

RESUMO

Sex steroid hormones are widely detected in molluscs and play important roles in sex determination, gonadal tissue maturation, and gametogenesis. Nevertheless, the signaling pathways of sex steroids in cephalopod have not yet been clearly elucidated. In the present study, a full-length sequence encoding the estrogen receptor (ER) was isolated from common Chinese cuttlefish, Sepiella japonica. The sjER cDNA clone was found to contain 1,788 nucleotides including a 1,470 bp open reading frame encoding 489 amino acid (aa) residues. The deduced ER protein consisted of six nuclear receptor characteristic domains. Based on a phylogenetic analysis, the ER DNA-binding domain and ligand-binding domain are highly conserved compared to other mollusc ERs. Highest aa identities were found for sjER with common octopus (Octopus vulgaris) ER (89%) and pacific oyster (Crassostrea gigas) ER (61%). Tissue expression analysis confirmed that sjER was widely distributed among tissues and predominantly expressed in the brain, liver, gonad (testis and ovary), and other accessory sexual gland (nidamental gland). The ER expression was temporally upregulated in the brain, liver, and ovary during the early sexual maturation period in S. japonica, which is coincident with the fluctuation of ovary estradiol content. These suggest that sjER may be involved in regulating the reproductive cycle of S. japonica. A fusion protein transient transfections assay showed that sjER was mainly located in the nucleus, suggesting a possible orthodox working mechanism of S. japonica ER in the nucleus through a ligand-dependent activation of specific gene transcription.


Assuntos
Decapodiformes/metabolismo , Receptores Estrogênicos/genética , Animais , Decapodiformes/genética , Feminino , Perfilação da Expressão Gênica , Masculino , Especificidade de Órgãos , Ovário/metabolismo , Filogenia , Domínios Proteicos , Receptores Estrogênicos/metabolismo , Reprodução , Testículo/metabolismo
18.
Ecol Evol ; 6(10): 3311-24, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27103988

RESUMO

Climate oscillations are the key factors to understand the patterns in modern biodiversity. East Asia harbors the most diverse temperate flora, largely because an extensive terrestrial ice cap was absent during repeated Pleistocene glaciation-interglacial cycles. Comparing the demographic histories of species that are codistributed and are close relatives may provide insight into how the process of climate change influences species ranges. In this study, we compared the spatial genetic structure and demographic histories of two coexisting Eleutherococcus species, Eleutherococcus senticosus and E. sessiliflorus. Both species are distributed in northern China, regions that are generally considered to be sensitive to climatic fluctuations. These regions once hosted temperate forest, but this temperate forest was replaced by tundra and taiga forest during the Last Glacial Maximum (LGM), according to pollen records. Using three chloroplast DNA fragments, we assessed the genetic structure of 20 and 9 natural populations of E. senticosus and E. sessiliflorus, respectively. Extremely contrasting genetic patterns were found between the two species; E. sessiliflorus had little genetic variation, whereas E. senticosus had considerably higher levels of genetic variation (15 haplotypes). We speculated that a recent severe bottleneck may have resulted in the extremely low genetic diversity in E. sessiliflorus. In E. senticosus, populations in Northeast China (NEC) harbored all of the haplotypes found in this species and included private haplotypes. The populations in NEC had higher levels of genetic diversity than did those from North China (NC). Therefore, we suggest that both the NC and NEC regions can sustain LGM refugia and that lineage admixture from multiple refugia took place after the LGM elevated the local genetic diversity in NEC. In NEC, multiple genetic hot spots were found in the Changbai Mountains and the Xiaoxing'an Range, which implied that multiple locations in NEC may sustain LGM refugia, even in the Xiaoxing'an Range.

19.
Sci Rep ; 6: 20110, 2016 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-26822920

RESUMO

Pharmacological activities of some natural products diminish and even disappear after purification. In this study, we explored the mechanisms underlying the decrease of acute oral toxicity of Coptidis Rhizoma extract after purification. The water solubility, in vitro absorption, and plasma exposure of berberine (the major active compound) in the Coptidis Rhizoma extract were much better than those of pure berberine. Scanning electron microscopy, laser scanning confocal microscopy (LSCM), and dynamic light scattering experiments confirmed that nanoparticles attached to very fine precipitates existed in the aqueous extract solution. The LSCM experiment showed that the precipitates were absorbed with the particles by the mouse intestine. High-speed centrifugation of the extract could not remove the nanoparticles and did not influence plasma exposure or acute oral toxicity. However, after extract dilution, the attached precipitates vanished, although the nanoparticles were preserved, and there were no differences in the acute oral toxicity and plasma exposure between the extract and pure berberine. The nanoparticles were then purified and identified as proteinaceous. Furthermore, they could absorb co-dissolved berberine. Our results indicate that naturally occurring proteinaceous nanoparticles in Coptidis Rhizoma extract act as concentration-dependent carriers that facilitate berberine absorption. These findings should inspire related studies in other natural products.


Assuntos
Berberina , Medicamentos de Ervas Chinesas , Nanopartículas , Proteínas de Plantas , Animais , Berberina/farmacocinética , Berberina/toxicidade , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/toxicidade , Camundongos , Nanopartículas/química , Nanopartículas/toxicidade , Proteínas de Plantas/química , Proteínas de Plantas/toxicidade
20.
Pharm Biol ; 54(6): 984-92, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26459663

RESUMO

CONTEXT: Fungal infections caused by fluconazole-resistant Candida albicans are an intractable clinical problem, calling for new efficient antifungal drugs. Kaempferol, an active flavonoid, has been considered a potential candidate against Candida species. OBJECTIVE: This work investigates the resistance reversion of kaempferol in fluconazole-resistant C. albicans and the underlying mechanism. MATERIALS AND METHODS: The antifungal activities of fluconazole and/or kaempferol were assessed by a series of standard procedures including broth microdilution method, checkerboard assay and time-kill (T-K) test in nine clinical strains as well as a standard reference isolate of C. albicans. Subsequently, the morphological changes, the efflux of rhodamine 6G, and the expressions of CDR 1, CDR 2, and MDR 1 were analysed by scanning electron microscope (SEM), inverted fluorescence microscope and quantitative reverse transcription polymerase chain reaction (qRT-PCR) in C. albicans z2003. RESULTS: For all the tested C. albicans strains, the minimum inhibitory concentrations (MICs) of fluconazole and kaempferol ranged 0.25-32 and 128-256 µg/mL with a range of fractional inhibitory concentration index of 0.257-0.531. In C. albicans z2003, the expression of both CDR 1 and CDR 2 were decreased after exposure to kaempferol alone with negligible rhodamine 6G accumulation, while the expression of CDR 1, CDR 2 and MDR 1 were all decreased when fluconazole and kaempferol were used concomitantly with notable fluorescence of rhodamine 6G observed. DISCUSSION AND CONCLUSION: Kaempferol-induced reversion in fluconazole-resistant C. albicans might be likely due to the suppression of the expression of CDR1, CDR2 and MDR1.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Candida albicans/efeitos dos fármacos , Farmacorresistência Fúngica/efeitos dos fármacos , Fluconazol/farmacologia , Proteínas Fúngicas/antagonistas & inibidores , Quempferóis/farmacologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Candida albicans/metabolismo , Farmacorresistência Fúngica/genética , Sinergismo Farmacológico , Fluconazol/administração & dosagem , Proteínas Fúngicas/genética , Expressão Gênica/efeitos dos fármacos , Quempferóis/administração & dosagem , Proteínas de Membrana Transportadoras/genética , Testes de Sensibilidade Microbiana
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