Phenolic compounds from Chaenomeles speciosa alleviate inflammation in lipopolysaccharide-treated RAW264.7 macrophages via the NF-κB and MAPK pathways

Chaenomeles speciosa(Sweet)Nakai cultivated widely in temperate regions possesses anti-inflammatory properties,however,the underlying molecular mechanisms remain not fully understood.In this study,a purified phenolic extract of C.speciosa rich in chlorogenic acid,procyanidin B1 and catechin(determined by HPLC-Q-TOF-MS/MS)exhibited dose-dependent anti-inflammatory effects on lipopolysaccharide(LPS)-treated RAW264.7 macrophages.The extract at 30μg/mL was most potent and enabled most cells in normal morphology under LPS stimulation without causing cytotoxicity.The extract suppressed the levels of nitric oxide(NO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β),and the mRNA and protein expressions of nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2).The mechanisms underlying such anti-inflammatory actions included the regulation of phosphorylation of related proteins to monitor the expressions of inflammatory mediators and genes in the nuclear factor kappa-B(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways.Therefore,the phenolic extract from C.speciosa is a desirable anti-inflammatory agent for inflammatory conditions to meet the rising demand for natural and cost-effective therapeutics.

The CsMYB123 and CsbHLH111 are involved in drought stress‑induced anthocyanin biosynthesis in Chaenomeles speciosa

Drought stress has been demonstrated to enhance the biosynthesis of anthocyanins in the leaves,resulting in an increased aesthetic appeal.However,the molecular mechanisms underlying drought-induced anthocyanin biosynthesis in Chaenomeles speciosa remain unclear.In this study,the metabolites of C.speciosa leaves were analyzed,and it was found that the content of cyanidin-3-O-rutinoside increased significantly under drought stress.The differentially expressed genes CsMYB123 and CsbHLH111 were isolated by transcriptomics data analysis and gene cloning,and gene overexpression and VIGS experiments verified that both play important roles in anthocyanin biosynthesis.Subsequently,Y1H and Dual-luciferase reporter assay showed that CsMYB123 binds to the promoters of anthocyanin biosynthesis-related structural genes(such as CsCHI,CsF3H,and CsANS),while CsbHLH111 was shown to bind to the promoter of CsCHI,positively regulating its activity.Furthermore,BIFC and Y2H assays unveiled potential protein–protein interactions between CsMYB123 and CsbHLH111 at the cell nucleus.Collectively,these results shed light on the critical roles played by CsMYB123 and CsbHLH111 in anthocyanin biosynthesis,thus providing a valuable insight into understanding the molecular mechanisms of how the MYB and bHLH genes regulate anthocyanin biosynthesis in the process of leaf coloration in C.speciosa.

Physicochemical and Functional Properties of Water Soluble Gum from Wrinkle Floweringquince(Chaenomeles Speciosa)Seeds

Wrinkle Floweringquince(Chaenomeles speciose(Sweet)Nakai)seed,as an unexploited forestry residue,contains considerable amount of bioactive carbohydrates with potential functionality,which was not widely concerned.The aim of this study is to determine the basic characterizations(molecular weight and functional group),specificcomponents(carbohydrate,protein and uronic acid contents),and functional properties of Chaenomeles speciosa seedgum(CSG).Results indicated that carbohydrate(63.80%),protein(13.69%)and uronic acid(10.30%)contents wereachieved.The CSG(average molecular weight,9.85×106 u)consists rhamnose,arabinose,xylose and glucose in a molarpercentage of 29.77꞉10.54꞉18.55꞉15.84,respectively.The Fourier transform infrared(FT-IR)analysis revealed hydroxyl,carboxyl and methyl groups andα-glycosidic linkages are founded in the CSG.The CSG was surface active and its abilityto decrease surface tension was comparable to commercial gums.Moreover,the CSG solutions showed pseudoplasticflow behavior under dynamic shear rate at high concentrations.The GSC also presented good emulsifying and foamingproperties,indicating the potential of the GSC as bioresource stabilizer and thickener in industry.

Mechanism of TSCS promote anti-Fas antibody-induced FLS apoptosis via blocking MC-tryptase-PAR-2-Rho-FLS signaling pathway

OBJECTIVE It has been supposed that mast cells have important participation in the physiopathology of RA,however,the role of mast cells in the pathogenesis of RA remains unclear.In this study,we observed the antiapoptotic effects of tryptase released by mast cell on RA synovial fibroblasts.METHODS Mast cells and fibroblasts synovial were obtained from mouse.Chemical mediator release was assessed by measuringβ-hexosa-minidase release.TSCS and bone marrow-derived mast cells were co-cultured;the toxic effects of TSCS on mast cell was measured by MTT and CCK-8 method;the releasing amount of tryptase in cell supernatants was measured by Elisa assay;the expression of FLS cell membrane receptor PAR-2 was detected by flow cytometry;the apoptosis of FLS cell was detected through flow cytometry and Western blotting;the level of activated Rho-GTP was detected by the pull-down method and Western blotting.RESULTS In this study,we observed the antiapoptotic effects of tryptase released by mast cell on RA synovial fibroblasts,and found that tryptase significantly increased the expression of PAR-2 on the surface of fibroblast-like synovial cell,significantly activated Rho kinase,significantly inhibited apoptosis of fibroblast-like synovial cell induced by CH11.The release rates ofβ-hexosaminidase and the level of tryptase from bone marrow-derived mast cells after stimulation with different antigen and co-cultured with TSCS significantly decreased compared to the control group.In the co-culture system of mast cells and fibroblast-like synovial cells,TSCS treatment significantly inhibited Rho kinase(P<0.05),significantly promoted apoptosis of fibroblast-like synovial cell induced by CH11(P<0.05).CONCLUSION These results demonstrate that tryptase may play a key role in the physiopathology of RA.TSCS can inhibit mast cells activation and promote FLS cells apoptosis.This provide theoretical and experimental basis for the study of mast cells as targets for new anti-RA drugs.

基于ISSR分子标记的皱皮木瓜遗传多样性分析

为了研究不同产地皱皮木瓜遗传多样性和亲缘性关系,本研究利用10个ISSR多态性引物对12份全国皱皮木瓜主产区的木瓜种质资源进行多态性和聚类分析,结果显示,各引物多态性百分率范围为65%~100%,皱皮木瓜种质资源的遗传相似系数在0.5057-0.8621之间,各种质间遗传信息较为丰富。聚类分析将12份种质划分为3个组,第1组又分为2个亚组,其中M4(湖南省桑植县-1)和M7(湖南省桑植县-2)亲缘关系最近,M1(云南省洱源县)和M10(湖北省郧西县)亲缘关系最远。以上结果表明12份全国皱皮木瓜主产区的木瓜种质资源具有较高的遗传多样性。

Microwave-low-pressure process(MWLP):An effective technology applied in extraction of total polyphenols

The microwave-low-pressure process(MWLP)of total polyphenols from Chaenomeles speciosa fruit was studied,and the advantages of MWLP were further evaluated by comparing with ultra high pressure(UHP)and microwave-assisted extraction(MAE).The influences of liquid/solid ratio,extraction time,pressure,and ethanol concentration on the performance of MWLP were investigated.Thereafter,the interactive variables were further optimized by the stepwise multiple quadratic regression model on the basis of the previous univariate analysis.The results showed that temperature as an intermediate variable in MWLP significantly affected the yields of polyphenols and 3-o-caffeoyl-quinic acid,which was determined by pressure and ethanol concentration.The optimized parameters were proved to be valid because the results predicted by the stepwise multiple quadratic regression model equations fit well with the experimental results.Compared with UHP,the predominance of MWLP was that the extraction time was shortened and the cost of extraction equipment was lowered.MWLP is an effective technology since MWLP was superior to MAE based on extraction yield,solvent loss and reproducibility.