Angiogenesis function of astragaloside Ⅳ in rats with myocardial infarction via PKD1-HDAC5-VEGF pathway

OBJECTⅣE This study aimed to investigate the role and mechanism of Astragaloside Ⅳ(AS-Ⅳ) in rats with myocardial infarction.METHODS The myocardial infarction model was established by ligation of the left anterior descending artery.The rats were randomly divided into sham,DMSO,model group,AS-Ⅳ and CID755673 groups.The rats were sacrificed 4 weeks later,and segmental heart samples were used for hematoxylin and eosin staining and masson staining.The expression of PKD1,HDAC5 and VEGF were analyzed using immunohistochemistry,reverse transcription poly.merase chain reaction and western blot.RESULTS Compared with the sham operation and DMSO groups,morphology of myocardium in model group was disordered,accompanied with necrotic myocar.dial cells and obvious collagen tissues.After treatment with AS-Ⅳ,the morphology of myocardium was obviously improved,and the number of new blood vessels increased significantly.However,after treatment with CID755673,the myocardial tissue of rats became disordered again,the necrotic cells increased,and some vessels closed.The expression levels of PKD1,HDAC5 and VEGF mRNA and protein in myocardial tissue of model group were significantly lower than the other four groups(P<0.05),whereas these levels in the AS-Ⅳ group were significantly higher than those in the other four groups(P<0.01).Additionally,the CID755673 group had significantly higher levels of PKD1,HDAC5 and VEGF mRNA and protein than the sham group,DMSO group and model group(P<0.05).CONCLUSION AS-Ⅳ may partly promote the angiogenesis of myocardial tissue in rats with myocardial infarction via the PKD1-HDAC5-VEGF pathway.

Guizhi Fuling Wan reverses drug resistance by regulating PTEN and MTDH in ovarian cancer SKOV3/DDP Cells

Abscract:OBJECTIVE To detect the reversal effect of Guizhi Fuling Wan on cisplatin-resistant ovarian cancer SKOV3/DDP cells and its relationship with protein expression of phosphatase and tensin homolog(PTEN) and metadherin(MTDH).METHODS Guizhi Fuling Wan(GFW) concentrated solution was prepared according to the Chinese Pharmacopoeia 2015 edition,Wistar rats were given GFW viagavage at 4 g·kg^(-1)·d^(-1),8 g·kg^(-1)·d^(-1),16 g·kg^(-1)·d^(-1),or given saline as blank control for 5 days.Blood samples were taken and the corresponding drug-containing low-dose sera,medium-dose sear,highdose sera and blank sera were prepared.The XCELLigence RTCA S16 real-time label-free cell analyzer was used to detect the reversal effect by the sera combined with cisplatin or paclitaxel in SKOV3/DDP cells.Annexin V-FITC and PI double-staining were used to detect the apoptosis-inducing effect of the sera in the cells.RT-qPCR and western blot were used to detect the mRNA and protein expression of PTEN and MTDH after the cells treated with the sera.RESULTS The inhibition rate of low-dose sera against SKOV3/DDP cells was less than 5%.After the low-dose sera combined with cisplatin or pacli.taxel,the IC50 of SKOV3/DDP cells against cisplatin and paclitaxel decreased by 3.01 and 1.79-fold,respectively.The total apoptosis rates induced by the low-dose sera,medium-dose sear,high-dose sera and blank sera in SKOV3/DDP cells were 11.08±0.13,19.42±0.30,24.23±0.31,and 3.21±0.24,respec.tively;there was a significant difference between the groups(P<0.01).RT-qPCR results showed that,compared with the blank serum,the sera can up-regulate the expression of PTEN mRNA and downregulate the expression of MTDH mRNA in a dose-dependent manner.Western blot results showed that the induction effect to PTEN protein and the inhibition effect to MTDH protein by the sera were gradually enhanced with thesera dose increasement.CONCLUSION The resistance reversal effect of Guizhi Fuling Wan on ovarian cancer SKOV3/DDP cells may be related to the inhibition of MTDH,up-regulation of PTEN and induction of apoptosis,providing with an experiment basis for the applica.tion of Guizhi Fuling Wan as a reversal agent for chemotherapy resistance of ovarian cancer.

Downregulated nuclear lncRNA NRON inhibits SHP2/Wnt/β-catenin signaling and cardiomyocyte differentiation during the development of Tetralogy of Fallot

Tetralogy of Fallot(TOF)is the most common cyanotic congenital heart disease and the incidence of late cardiac death in long-term survivors continues to increase.1 So,there is an urgent need to explore the etiology and pathogenesis of TOF.The precise cause of TOF is currently unclear,and exploration of the pathogenesis has focused increasingly in recent years on the roles of noncoding gene products,especially long noncoding RNAs(lncRNAs).

Prediction and verifcation of the key ingredients and molecular targets of Guizhi Fuling capsule against tumour metastasis and resistance

Purpose:The well-known traditional Chinese formula Guizhi Fuling capsule(GFC)has been reported to reverse ovarian cancer drug resistance.Extrachromosomal DNA(ecDNA)plays an important role in tumour metastasis and resistance.The purpose of this study was to investigate the potential mechanisms by which GFC blocks tumour metastasis and reverses drug resistance by targeting ecDNA.Methods:CNKI and PubMed were used to obtain pharmacokinetic research data on GFC in rats,and the bioactive ingredients detected in rat serum or plasma were collected.Network databases were used to screen the abnormally expressed genes in ecDNA,tumour metastasis genes,resistance genes,and the active ingredient targets of GFC.The KOBAS3.0 database was used to enrich the KEGG pathways and GO functions;the STRING platform was used to construct the core protein interaction network;and the molecular docking online tool SwissDock was used to analyse the binding activity of the core targets and the active ingredients.RT-qPCR,Western blotting and laser confocal microscopy were used to verify the efect of the sera containing GFC on ecDNA,mRNA and protein expression of key targets.Results:Twenty-three bioactive ingredients of GFC were retrieved from PubMed and CNKI.Nine shared targets were simultaneously involved in abnormal genes in ecDNA,tumour metastasis and resistance and the active ingredient targets of GFC.GO functional analysis indicated that the cotargets involved cell proliferation,apoptotic regulation,nuclear functions,etc.The potential pathways involved in the reversal of tumour metastasis and drug resistance of GFC were the PI3K-Akt signalling,cancer,and platinum drug resistance pathways.Three shared proteins targeting ecDNA(AKT1,EGFR and MYC)stand out from the top 20 PPI targets,and all of the bioactive ingredients of GFC have strong binding afnity to the three proteins.The active ingredients can reduce the expression of MYC,EGFR and AKT1 mRNA and protein and the amount of ecDNA in drug-resistant OC cells.Conclusions:GFC targeting ecDNA to reverse tumour metastasis and drug resistance has the characteristics of multiple ingredients,multiple targets,and multiple pathways,which provides a new perspective for the development of new drugs targeting ecDNA to beneft tumour treatment.