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1.
Article in Chinese | WPRIM | ID: wpr-771507

ABSTRACT

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.


Subject(s)
Adenosine Triphosphatases , Genetics , Candida albicans , Drugs, Chinese Herbal , Pharmacology , Fungal Proteins , Genetics , Gene Expression Regulation, Fungal , HSP70 Heat-Shock Proteins , Genetics , Hyphae , Microscopy, Electron, Scanning
2.
Article in Chinese | WPRIM | ID: wpr-335817

ABSTRACT

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.

3.
Article in Chinese | WPRIM | ID: wpr-330244

ABSTRACT

<p><b>OBJECTIVE</b>To investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.</p><p><b>METHOD</b>Serial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.</p><p><b>RESULT</b>The MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.</p><p><b>CONCLUSION</b>EAHD could effectively inhibit adherence of C. glabrata.</p>


Subject(s)
Acetates , Candida glabrata , Physiology , Drugs, Chinese Herbal , Pharmacology , Fungal Proteins , Genetics , Lectins , Genetics , Plant Extracts , Pharmacology
4.
Article in Chinese | WPRIM | ID: wpr-330173

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of butyl alcohol extract of Baitouweng decoction ( BAEB) on yeast-to-hyphae transition of Candida albicans isolates from vulvovaginal candidiasis (VVC) in alkaline pH.</p><p><b>METHOD</b>Serial 2-fold dilution assay was used to determine the minimal inhibitory concentrations (MICs) of Baitouweng decoction extracts against C. albicans isolates from VVC, XTT assay was applied to determine the metabolic activity of C. albicans hypha treated by BAEB for 6 h. The morphological change of C. albicans treated by BAEB was inspected at different pH by inverted microscope, fluorescence microscope, scanning electron microscopy (SEM). Solid agar plate and semi-solid agar were utilized to evaluate colony morphology and invasive growth of C. albicans, respectively. Quantitative Real-time PCR (qRT-PCR) was adopted to observe the expressions of hyphae-specific genes including HWP1, ALS3, CSH1, SUN41 and CaPDE2.</p><p><b>RESULT</b>The MIC of BAEB against C. albicans is less than that of other extracts; hyphae grow best at pH 8. 0; 512 mg · L(-1) and 1,024 mg · L(-1) BAEB could inhibit formation of hyphae and influence colony morphology. When treated by 512 mg · L(-1) and 1,024 mg · L(-1) BAEB, the colonies became smooth; while by 0 and 256 mg · L(-1) BAEB, the colonies became wrinkled. In semi-solid agar, the length of hyphae decreased steadily as the concentration of BAEB lowered. The expression of HWP1, ALS3, CSHl, SUN41 were downregulated by 5.12, 4.26, 3.2 and 2.74 folds, and CaPDE2 was upregulated by 2.38 fold.</p><p><b>CONCLUSION</b>BAEB could inhibit yeast-to-hyphae transition of C. albicans isolates from VVC in alkaline pH.</p>


Subject(s)
Antifungal Agents , Pharmacology , Candida albicans , Genetics , Candidiasis, Vulvovaginal , Drug Therapy , Microbiology , Drugs, Chinese Herbal , Pharmacology , Female , Humans , Hydrogen-Ion Concentration , Hyphae
5.
Article in Chinese | WPRIM | ID: wpr-330170

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.</p><p><b>METHOD</b>The minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.</p><p><b>RESULT</b>MICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.</p><p><b>CONCLUSION</b>The combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.</p>


Subject(s)
Antifungal Agents , Pharmacology , Candida albicans , Metabolism , Drug Resistance, Fungal , Drug Synergism , Drugs, Chinese Herbal , Pharmacology , Ergosterol , Fluconazole , Pharmacology , Microbial Sensitivity Tests
6.
Article in Chinese | WPRIM | ID: wpr-305306

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of andrographolide (AG) on quroum sensing (QS) and relevant virulence genes of Candida albicans.</p><p><b>METHOD</b>Gas-chromatography-mass spectrometry (GC-MS) was applied to detect the changes in the content of farnesol and tyrosol in C. albicans intervened by AG. The real-time quantitative PCR (qRT-PCR) was adopted to inspect the expressions of relevant virulence genes such as CHK1, PBS2 and HOG1 regulated by QS.</p><p><b>RESULT</b>At 2 h after the growth of C. albican, the farnesol and tyrosol secretions reduced, without notable change after intervention with AG. The secretions were highest at 12 h and decreased at 24 h. After the intervention with different concentrations of AG, the farnesol content reduces, whereas tyrosol increased, indicating a dose-dependence, particularly with 1 000 mg x L(-1) AG. qRT-PCR revealed that 1 000 mg x L(-1) AG could down-regulate CHK1 by 2.375, 3.330 and 4.043 times and PBS2 by 2.010, 4.210 and 4.760 times, with no significant change in HOG1.</p><p><b>CONCLUSION</b>AG could inhibit the farnesol secretion, promote the tyrosol secretion and down-regulate QS-related virulence genes CHK1 and PBS2 expressions.</p>


Subject(s)
Candida albicans , Genetics , Physiology , Diterpenes , Pharmacology , Farnesol , Metabolism , Gas Chromatography-Mass Spectrometry , Genes, Fungal , Phenylethyl Alcohol , Metabolism , Quorum Sensing , Real-Time Polymerase Chain Reaction , Virulence , Genetics
7.
Article in Chinese | WPRIM | ID: wpr-244568

ABSTRACT

Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.


Subject(s)
Anti-Inflammatory Agents , Pharmacology , Biofilms , Candida albicans , Genetics , Physiology , Diterpenes , Pharmacology , Dose-Response Relationship, Drug , Fungal Proteins , Genetics , Gene Expression Regulation, Fungal , HSP90 Heat-Shock Proteins , Genetics , Microscopy, Electron, Scanning , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
8.
Article in Chinese | WPRIM | ID: wpr-311003

ABSTRACT

In recent years, with the constant increase in the population with hypoimmunity, bacterial and fungal infections have been increasing. Due to the drug resistance, clinically optional anti-bacterial and antifungal medicines become increasingly limited. Scutellaria baicalensis, a species of perennial herbaceous plant of scutellaria genus of lamiaceae family, and its effective components have multiple pharmacological effects such as anti-inflammation, anti-oxidation, anti-tumor, anti-microbial. Especially, its remarkable antibacterial and antifungal activities are of great significance to treat the increasing number of cases with drug-fast bacterial and antifungal infections. In this paper, the authors summarized the advance in studies on antibacterial and antifungal effects and mechanisms in recent years on the basis of the domestic and foreign studies on S. baicalensis and its effective ingredients.


Subject(s)
Animals , Anti-Infective Agents , Chemistry , Pharmacology , Bacterial Infections , Drug Therapy , Microbiology , Humans , Mycoses , Drug Therapy , Microbiology , Plant Extracts , Chemistry , Pharmacology , Scutellaria , Chemistry , Scutellaria baicalensis , Chemistry
9.
Article in Chinese | WPRIM | ID: wpr-327866

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.</p><p><b>METHOD</b>The rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.</p><p><b>RESULT</b>The YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.</p><p><b>CONCLUSION</b>YHN could inhibit C. albicans biofilms in rats.</p>


Subject(s)
Animals , Biofilms , Candida albicans , Cell Biology , Physiology , Catheters , Microbiology , Cell Adhesion , Diterpenes , Chemistry , Pharmacology , Dose-Response Relationship, Drug , Rats
10.
Article in Chinese | WPRIM | ID: wpr-321323

ABSTRACT

<p><b>OBJECTIVE</b>To observe the inhibitory effect of different extract fractions from Longdan Xiegan decoction on biofilms of Candida albicans, and discuss its possible mechanism.</p><p><b>METHOD</b>The micro-dilution method and the XTT reduction assay were adopted to explore the antifungal activity of different extract fractions from Longdan Xiegan decoction and detect the inhibitory effect of different extracts on biofilms of C. albicans. The expression quantity of the adhesion related gene ALS1 and hypha formation SUN41 were detected by qRT-PCR.</p><p><b>RESULT</b>The MICs of extracts from Longdan Xiegan decoction, including petroleum ether, water, butanol, methanol and ethyl acetate, against C. albicans were > 1 000, > 1 000, > 1 000, 125, 125 mg x L(-1). The SMIC50 against biofilms of C. albicanswere > 1 000, > 1000, > 1 000, 500, 500 mg x L(-1). The SMIC50 were > 1 000, > 1 000, > 1 000, > 1 000 and 1 000 mg x L(-1). 1 000 mg x L(-1) ethyl acetate extracts could considerably inhibit the expression of the adhesion related gene ALS1 and hypha formation SUN41.</p><p><b>CONCLUSION</b>The ethyl acetate extract showed the greatest activity against Candida albicans biofilms.</p>


Subject(s)
Antifungal Agents , Pharmacology , Biofilms , Candida albicans , Candidiasis , Drug Therapy , Microbiology , Drugs, Chinese Herbal , Pharmacology , Humans , Hyphae , Microbial Sensitivity Tests
11.
Article in Chinese | WPRIM | ID: wpr-341807

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on hyphae development of Candida albicans.</p><p><b>METHOD</b>Inverted microscope, fluorescence microscope, SEM were applied to inspect the Morphological change of C. albicans treated by EAHD at different concentrations. Solid agar plate was utilized to evaluate the colony morphology. Quantitative Real-ime PCR(qRT-PCR) was adopted to observe the expression of hyphae-specific genes such as HWP1, ALS3, UME6, CSH1, SUN41, CaPDE2.</p><p><b>RESULT</b>EAHD with concentration of 312 and 1 250 mg . L-1 could inhibit formation of hyphae and colony morphology. The expression of HWP1, ALS3, UME6, CSH1 were downregulated 4. 13, 3. 64, 2. 46, 2. 75 folds ,while the expression of SUN41 were upregulated 7. 26 folds, CaPDE2 keep unchanged.</p><p><b>CONCLUSION</b>EAHD could inhibit formation of hyphae and colony morphologies of C. albicans through downregulating HWP1, ALS3, UME6 and CSH1.</p>


Subject(s)
Acetates , Biofilms , Candida albicans , Cell Biology , Genetics , Down-Regulation , Drugs, Chinese Herbal , Pharmacology , Fungal Proteins , Genetics , Gene Expression Regulation, Fungal , Hyphae , Medicine, Chinese Traditional , Microscopy, Fluorescence , Reverse Transcriptase Polymerase Chain Reaction
12.
Article in Chinese | WPRIM | ID: wpr-319697

ABSTRACT

Andrographolide (AG) is a main effective ingredient in leaves of Andrographis paniculata. AG and its derivatives have such effects in anti-infection, anti-tumor and immuno-regulation. Particularly, its antibacterial, antiviral and anti-protozoal activities play an increasingly important role in resisting opportunistic infections. On the basis of our studies on AG over the years, we made a summary for the basis and clinical studies on AG and its derivatives over the past 10 years.


Subject(s)
Andrographis , Chemistry , Animals , Anti-Infective Agents , Chemistry , Pharmacology , Diterpenes , Chemistry , Pharmacology , Drugs, Chinese Herbal , Chemistry , Pharmacology , Humans , Structure-Activity Relationship
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