Effects of ginkgo flavone aglycone on atherosclerosis based on network pharmacology,molecular docking,and in vitro experiments

Background:Ginkgo flavone aglycones(GA),a Ginkgo(Ginkgo biloba)extract,has been proven to have good biological activity in atherosclerosis(AS)treatment.Moreover,its active compounds and the corresponding mechanism for the treatment of AS remain unclear.Methods:To evaluate and identify the potential pharmacological mechanisms of GA in AS treatment,the program Cytoscape was used to generate network mappings of the GA-AS-potential target gene.GO and KEGG enrichment analyses were performed to further investigate the potential mechanism of AS and the pharmacological properties of GA.A molecular docking approach was utilized to determine the GA components that interact with Akt.In vitro experiments were carried out to identify the anti-atherosclerotic effects of GA by targeting Akt.Results:Network pharmacological research determined that the active components of GA(quercetin,kaempferol,and isorhamnetin)correlated with AS target genes such as AKT1,EGFR,SRC,ESR1,PTGS2,MMP9,KDR,GSK3B,APP,and MMP2,respectively.GO enrichment and KEGG analysis showed that PI3K-Akt signaling may play an important role in GA treatment.Molecular docking experiments indicated that quercetin,kaempferol,and isorhamnetin integrate into the binding pockets of the most potentially beneficial GA-AS target protein(Akt).Consequently,cell experiments were conducted to support the anti-atherosclerotic activity of GA on AS by inhibiting the phosphorylation of AKT1 and its downstream signaling molecules,which regulated the proliferation of HASMCs.Conclusion:Our results detailed GA's active ingredients,potential targets,and molecular basis against AS.GA may exert anti-atherosclerotic effects by suppressing Akt phosphorylation and inhibiting the proliferation of HASMCs.It also proposed a viable approach to determining the scientific foundation and therapeutic mechanism of Chinese herbal medicine extracts in disease therapy.

Fidgetin knockdown and knockout influences female reproduction distinctly in mice

Microtubule-severing proteins(MTSPs),are a family of proteins which use adenosine triphosphate to sever microtubules.MTSPs have been shown to play an important role in multiple microtubule-involved cellular processes.One member of this family,fidgetin(FIGN),is also involved in male fertility;however,no studies have explored its roles in female fertility.In this study,we found mouse fidgetin is rich within oocyte zona pellucida(ZP)and is the only MTSP member to do so.Fidgetin also appears to interact with all three ZP proteins.These findings prompted us to propose that fidgetin might prevent polyspermy.Results from in vitro maturation oocytes analysis showed that fidgetin knockdown did cause polyspermy.We then deleted all three fidgetin isoforms with CRISPR/Cas9 technologies;however,female mice remained healthy and with normal fertility.Of all mouse MTSPs,only the mRNA level of fidgetin-like 1(FIGNL1)significantly increased.Therefore,we assert that fidgetin-like 1 compensates fidgetin's roles in fidgetin knockout female mice.

ATP50 Hypo-crotonylation Caused by HDAC2 Hyper-Phosphorylation Is a Primary Detrimental Factor for Downregulated Phospholipid Metabolism under Chronic Stress

Objective.Chronic stress(CS)-induced abnormal metabolism and other subsequent aspects of abnormality are threatening human health.Little is known regarding whether and how protein post-translational-modifications(PTMs)correlate with abnormal metabolism under CS.The aim of this study was to address this issue and also identify novel key protein PTM.Methods.First,we screened which pan-PTM had significant change between control and CS female mice and whether clinical CS females had similar pan-PTM change.Second,we performed quantitative PTM-omics and metabolomics to verify the correlation between abnormal protein PTMs and atypical metabolism.Third,we performed quantitative phospho-omics to identify the key PTM-regulating enzyme and investigate the interaction between PTM protein and PTM-regulating enzyme.Fourth,we attempted to rectify the abnormal metabolism by correcting the activity of the PTM-regulating enzyme.Finally,we examined whether the selected key protein was also correlated with stress scores and atypical metabolism in clinical women.Results.We initially found that multiple tissues of CS female mice have downregulated pan-crotonylation,and verified that the plasma of clinical CS females also had downregulated pan-crotonylation.Then we determined that ATP5O-K51 crotonylation decreased the most and also caused gross ATP5O decrement,whereas the plasma of CS mice had downregulated phospholipids.Next,downregulating ATP5O crotonylation partially recapitulated the downregulated phospholipid metabolism in CS mice.Next,we verified that HDAC2-S424 phosphorylation determined its decrotonylation activity on ATP5O-K51.Furthermore,correcting HDAC2 hyper-phosphorylation recovered the gross ATP5O level and partially rescued the downregulated phospholipid metabolism in CS mice.Finally,the ATP5O level was also significantly iower and correlated with high stress scores and downregulated phospholipid metabolism in clinical female plasma.Conclusion.This study discovered a novel PTM mechanism involving two distinct types of PTM in CS and provided a novel reference for the clinical precautions and treatments of CS.

PNAS:科学家发现新的精子运动调节器

南京医科大学生殖医学国家重点实验室、基础医学院组织胚胎学系刘明兮课题组与贝勒医学院MartinM．Matzuk课题组、大阪大学MasahitoIkawa课题组等共同解析了一个精子运动调节基因TCTE1，结果发表于《美国科学院院刊》（PNAs）。

Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

The use of two inhibitors of Mek1/2 and Gsk3β(2i)promotes the generation of mouse diploid and haploid embryonic stem cells(ESCs)from the inner cell mass of biparental and uniparental blastocysts,respectively.However,a system enabling long-term maintenance of imprints in ESCs has proven challenging.Here,we report that the use of a two-step a2i(alternative two inhibitors of Src and Gsk3β,TSa2i)derivation/culture protocol results in the establishment of androgenetic haploid ESCs(AG-haESCs)with stable DNA methylation at paternal DMRs(differentially DNA methylated regions)up to passage 60 that can efficiently support generating mice upon oocyte injection.We also show coexistence of H3K9me3 marks and ZFP57 bindings with intact DMR methylations.Furthermore,we demonstrate that TSa2itreated AG-haESCs are a heterogeneous cell population regarding paternal DMR methylation.Strikingly,AGhaESCs with late passages display increased paternal-DMR methylations and improved developmental potential compared to early-passage cells,in part through the enhanced proliferation of H19-DMR hypermethylated cells.Together,we establish AG-haESCs that can longterm maintain paternal imprints.

Effect of endostatin overexpression on angiotensin Ⅱ-induced cardiac hypertrophy in rats

Background:Endostatin,a biologically active fragment of collagen XVIII,has been observed in patients w让h ischemic heart disease.The aim of the present study was to investigate whether endostatin overexpression could attenuate cardiac hypertrophy by inhibiting the cyclic adenosine monophosphate-protein kinase A(cAMP-PKA)signaling pathway.Methods:This study was examined in vivo in rats and in vitro in primary neonatal rat cardiomyocytes treated with angiotensin(Ang)II to model cardiac hypertrophy.Twenty-four male Sprague-Dawley rats were randomized into adenovirus(Ad)-green fluorescent protein,Ang II,Ad-endostatin,and Ang II+Ad-endostatin groups(w=6 in each group).Four weeks later,all the rats were weighed and sacrificed after transthoracic echocardiography.Cardiac function was evaluated by transthoracic echocardiography,cardiomyocyte size was evaluated by hematoxylin-eosin staining.Levels of atrial natriuretic peptide(ANP)and brain natriuretic peptide(BNP)were evaluated by quantitative reverse-transcription polymerase chain reaction or Western blotting,PKA level was evaluated by Western blotting,and cAMP level was evaluated by enzyme-linked immunosorbent assay.Statistical significance among multiple groups was evaluated by one-way analysis of variance.Results:Endostatin overexpression reduced the increases in left ventricle(LV)mass(P=0.0063),LV mass/body weight(BW)(P=0.0013),interventricular septal thickness(IVS)in diastole(P=0.0013),IVS in systole(P=0.0056),left ventricular posterior wall thickness(LVPW)in diastole(P=0.0291),LVPW in systole(P=0.0080),heart weight(HW)(P=0.0138),HW/BW(P=0.0001),and HW/tibial length(P=0.0372)in Ang Il-treated rats.In addition,endostatin overexpression reduced cardiomyocyte cross-sectional area expansion,and reduced the levels of ANP and BNP in Ang Il-treated rats(P=0.0251 and 0.0477 for messenger RNA[mRNA]),and primary neonatal rat cardiomyocytes(P=0.0188 and P=0.0024 for mRNA;P=0.0023 and 0.0013 for protein,respectively).Additionally,endostatin overexpression reduced the increase of cAMP(P=0.0054)and PKA(P=0.0328)levels in cardiomyocytes treated with Ang II.Treatment with cAMP reversed the effects of endostatin overexpression on ANP(P=0.0263)and BNP(P=0.0322)levels in cardiomyocytes induced by Ang II.Conclusion:Endostatin overexpression could alleviate cardiac hypertrophy by inhibiting the cAMP-PKA signaling pathway.