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1.
Zhongguo Zhong Yao Za Zhi ; 44(13): 2709-2718, 2019 Jul.
Article in Chinese | MEDLINE | ID: mdl-31359681

ABSTRACT

To screen the active ingredients of Gardenia jasminoides and potential targets,and investigate the mechanisms against cholestasis based on network pharmacology technology. Twenty-one active components of G. jasminoides were retrieved and the target sites were screened by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform( TCMSP). Cytoscape3. 2. 1 was used to construct the component-target network. Two hundred and eight targets related to cholestasis were searched and screened through Dis Ge NET,KEGG and OMIM databases. The key targets of G. jasminoides components and cholestasis were integrated and screened,and the component-target-disease network was constructed with Cytoscape 3. 2. 1 software to screen out the core network whose freedom degree was greater than the average value. The Clue GO plug-in of Cytoscape 3. 2. 1 software was used to analyze the biological processes and pathway enrichment of G. jasminoides in regulation of cholestasis. GO biological process analysis revealed 17 biological processes,involving 3 signaling biological processes related to cholestasis,i.e. acute inflammatory response,positive regulation of reactive oxygen species metabolic process,and nitric oxide biosynthetic process. KEGG-KEEG-305 terms and REACTOME pathways analysis revealed 17 regulatory pathways,involving 4 signaling pathways related to cholestasis,i.e. metabolism of xenobiotics by cytochrome P450,nuclear receptor transcription pathway,GPVI-mediated activation cascade and platelet activation. It was found that aqueous extract of G. jasminoides could improve serum biochemical abnormalities in ANIT-induced cholestasis rats. Aqueous extract of G. jasminoides could decrease the protein and mRNA expression levels of ESR1 in liver tissues,and increase the protein and mRNA expression levels of PPARG,NOS2,F2 R,NOS3,and NR3 C1. To sum up,the possible mechanisms of G. jasminoides against cholestasis may be related with the above three processes and four pathways.


Subject(s)
Cholestasis/drug therapy , Drugs, Chinese Herbal/pharmacology , Gardenia/chemistry , Plant Extracts/pharmacology , Animals , Medicine, Chinese Traditional , Rats , Signal Transduction
2.
Zhongguo Zhong Yao Za Zhi ; 44(4): 787-795, 2019 Feb.
Article in Chinese | MEDLINE | ID: mdl-30989893

ABSTRACT

To investigate the effects of geniposidic acid( GPA) on hepato-enteric circulation in cholestasis rats,and to explore the mechanism based on the sirtuin 1( Sirt1)-farnesol X receptor( FXR) pathway,sixty SD rats were randomly divided into 6 groups:blank control group,ANIT model group,ursodeoxycholic acid group( 100 mg·kg~(-1)·d-1 UDCA),and GPA high,medium and low( 100,50 and 25 mg·kg~(-1)·d-1) dosage groups,10 rats in each group. Corresponding drugs were intragastrically( ig) administered for10 days. After administration on day 8,all rats except blank rats were administered with 65 mg·kg~(-1)α-naphthalene isothiocyanate( ANIT) once. After the last administration,the serum levels of alanine aminotransferase( ALT),glutamine oxalacetate aminotransferase( AST),gamma-glutamyltransferase( γ-GGT),alkaline phosphatase( ALP),total bilirubin( TB) and total bile acid( TBA)were measured,and the mRNA transcription levels of Sirt1,FXR,multidrug resistant associated protein 2( MRP2),bile salt export pump( BSEP),sodium taurocholate contractible polypeptide( NTCP) in liver and apical sodium bile acid transporter( ASBT),ileum bile acid binding protein( IBABP) in ileum were detected by reverse transcription-polymerase chain reaction( RT-PCR). The protein expression levels of Sirt1,FXR and NTCP were detected by Western blot; the expression of MRP2,BSEP in liver and ASBT,IBABP in ileum were determined by immunofluorescence three staining. Primary rat hepatocytes were cultured in vitro to investigate the inhibitory effect of GPA on a potent and selective Sirt1 inhibitor( EX 527),and the mRNA and protein expression levels of Sirt1 and FXR were detected by RT-PCR and Western blot. GPA significantly decreased the levels of ALT,AST,γ-GGT,ALP,TB,TBA in serum( P<0.01) and improved the pathological damage of liver tissues in ANIT-induced cholestasis rats; significantly increased the mRNA and protein expression levels of Sirt1,FXR,MRP2,BSEP,NTCP in liver and ASBT,IBABP in ileum( P< 0.01). In vitro primary hepatocytes experiment indicated that the gene and protein expression levels of FXR and Sirt1 were noticeably improved by GPA in primary hepatocytes inhibited by EX-527( P<0.01). It was found that the improvement of GPA was in a dose-dependent manner. GPA could improve bile acid hepatointestinal circulation and play a liver protection and cholagogu role in cholestasis rats induced by ANIT.The mechanism may be that GPA activated FXR by regulating Sirt1,a key regulator of oxidative stress injury,and then the activated FXR could regulate protein of bile acid hepato-enteric circulation.


Subject(s)
Cholestasis , Animals , Iridoid Glucosides , Liver , Rats , Rats, Sprague-Dawley , Receptors, Cytoplasmic and Nuclear , Signal Transduction , Sirtuin 1
3.
Zhong Yao Cai ; 39(9): 2046-8, 2016 Sep.
Article in Chinese | MEDLINE | ID: mdl-30209910

ABSTRACT

Objective: To establish a high phase liquid chromatography method of the content in quercetin,luteolin,apigenin,acacetin,and to compare the difference of content from four different varieties of Dendranthema morifolium in simultaneously. Methods: The UPLC methods were adopted,and the chromatographic column was Waters ACQUITYUPLC; the column was BEH C18( 50 mm ×2. 1 mm,1. 7 µm),the mobile phase was 0. 1% phosphoric acid solution-methanol in gradient elution,the flow rate was 0. 2 m L/min;and the detection wavelength was set at 320 nm; the column temperature 25 ℃; and the sample quantity was 1 µL. Results: In the range of 0. 0027 0. 0135 mg/m L( r1= 0. 9962) concentration within quercetin in a good linear relationship between peak area. In the range of0. 0032 0. 0160 mg/m L( r2= 0. 9963) concentration within luteolin in a good linear relationship between peak area. In the range of0. 0029 0. 0145 mg/m L( r3= 0. 9964) of apigenin in the mass concentration and the peak area. In the range of 0. 0029 0. 0145 mg/m L( r4= 0. 9963) concentration within acacetin in a good linear relationship between peak area. Conclusion: This method can be determined daisy quercetin,luteolin,apigenin,acacetin content in Dendranthema morifolium.


Subject(s)
Chromatography, High Pressure Liquid , Chrysanthemum , Drugs, Chinese Herbal , Flavonoids
4.
Int Immunopharmacol ; 28(1): 344-53, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26093272

ABSTRACT

In previous study, we identified that microRNA (miR)-152 expression was down-regulated in RA model rats, and overexpression of miR-152 inhibited the canonical Wnt signaling through the DNA methyltransferase (DNMT1) inhibition. However, the exact molecular mechanisms of DNMT1 were unclear. In this work, we investigate whether DNMT1 affects the pathogenesis of RA model rats and targets the miR-152 promoter. The effects of DNMT1 on the canonical Wnt signaling, the pathogenesis of RA model rats and the SFRP1 expression were detected by the real time qPCR, Western blotting, ELISA, MTT and viable cell number assay. The interaction between miR-152 and DNMT1, methyl CpG binding protein 2 (MeCP2) was investigated by real time qPCR and chromatin immunoprecipitation (ChIP). Our results revealed that increased DNMT1 activated the canonical Wnt signaling could not only by targeting SFRP4 may also by SFRP1 in RA model rats. Furthermore, treatment of DNMT1 inhibitor, 5-aza-2'-deoxycytidine (5-azadC), or knockdown of DNMT1, or knockdown of MeCP2 led to increased miR-152 expression by reversion of its promoter hypermethylation, DNMT1 and MeCP2 binding to the CpG islands of miR-152 promoter. Interestingly, it is proved a synergistic inhibition effect of DNMT1 and MeCP2 in this process. Moreover, overexpression of miR-152 could inhibit DNMT1 expression and result in a decrease of DNMT1 and MeCP2 binding to miR-152 promoter, and inhibition of miR-152 expression would reverse it. These observations demonstrate a crucial functional crosstalk between miR-152 and the DNMT1, MeCP2 by a double-negative circuit involved in the pathogenesis of RA model rats.


Subject(s)
Arthritis, Rheumatoid/metabolism , DNA (Cytosine-5-)-Methyltransferases/metabolism , Methyl-CpG-Binding Protein 2/metabolism , MicroRNAs/metabolism , Wnt Signaling Pathway , Animals , DNA (Cytosine-5-)-Methyltransferase 1 , DNA (Cytosine-5-)-Methyltransferases/genetics , Disease Models, Animal , Male , MicroRNAs/genetics , Promoter Regions, Genetic , Rats, Sprague-Dawley , Wnt Proteins/metabolism
5.
Zhongguo Zhong Yao Za Zhi ; 32(7): 623-6, 2007 Apr.
Article in Chinese | MEDLINE | ID: mdl-17583207

ABSTRACT

OBJECTIVE: To observe the effect of cyclovirobuxinum-D (CVB-D) on cerebral ischemia-reperfusion injury in rats and explore its mechanisms. METHOD: One hundred and twenty rats were randomly divided into three CVB-D groups (2, 1, 0.5 mg x kg(-1)), Nimodipine group (2 mg x kg(-1)), model group and sham operated group, 20 rats each group. Rat cerebral ischemia-reperfusion injury model was induced by middle cerebral artery occlusion, the nerve injury symptoms was evaluated, the level of SOD and MDA in brain tissue were determined, the concentration of intracellar Ca2+ of brain was measured, and the pathological change of brain was also observed. RESULT: CVB-D could improve the nerve injury symptoms, reduce the infarction area of brain, the concentration of intracellar Ca2+ and the level of MDA, increase the activity of SOD, and decrease the pathological change of brain. CONCLUSION: CVB-D has protective effect on cerebral ischemia-reperfusion injury in rats.


Subject(s)
Brain/drug effects , Drugs, Chinese Herbal/pharmacology , Neuroprotective Agents/pharmacology , Reperfusion Injury/prevention & control , Animals , Brain/metabolism , Brain/pathology , Buxus/chemistry , Calcium/metabolism , Drugs, Chinese Herbal/isolation & purification , Infarction, Middle Cerebral Artery/complications , Male , Malondialdehyde/metabolism , Neuroprotective Agents/isolation & purification , Plants, Medicinal/chemistry , Random Allocation , Rats , Rats, Sprague-Dawley , Reperfusion Injury/etiology , Reperfusion Injury/metabolism , Superoxide Dismutase/metabolism
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