Effects of 1-methylcyclopropene on skin greasiness and quality of'Yuluxiang'pear during storage at 20℃

During storage at 20℃,specific pear cultivars may exhibit a greasy texture and decline in quality due to fruit senescence.Among these varieties,‘Yuluxiang’is particularly susceptible to peel greasiness,resulting in significant economic losses.Therefore,there is an urgent need for a preservative that can effectively inhibit the development of greasiness.Previous studies have demonstrated the efficacy of 1-methylcyclopropene(1-MCP)in extending the storage period of fruits.We hypothesize that it may also influence the occurrence of postharvest peel greasiness in the‘Yuluxiang’pears.In this study,we treated‘Yuluxiang’pears with 1-MCP.We stored them at 20℃while analyzing the composition and morphology of the surface waxes,recording enzyme activities related to wax synthesis,and measuring indicators associated with fruit storage quality and physiological characteristics.The results demonstrate that prolonged storage at 20℃leads to a rapid increase in skin greasiness,consistent with the observed elevations in L^(*),greasiness score,and the content of total wax and greasy wax components.Moreover,there were indications that cuticular waxes underwent melting,resulting in the formation of an amorphous structure.In comparison to controls,the application of 1-MCP significantly inhibited increments in L^(*) values as well as grease scores while also reducing accumulation rates for oily waxes throughout most stages over its shelf period,additionally delaying transitions from flaky-wax structures towards their amorphous counterparts.During the initial 7 d of storage,several enzymes involved in the biosynthesis and metabolism of greasy wax components,including lipoxygenase(LOX),phospholipase D(PLD),andβ-ketoacyl-CoA synthase(KCS),exhibited an increase followed by a subsequent decline.The activity of LOX during early shelf life(0–7 d)and the KCS activity during middle to late shelf life(14–21 d)were significantly suppressed by 1-MCP.Additionally,1-MCP effectively maintained firmness,total soluble solid(TSS)and titratable acid(TA)contents,peroxidase(POD),and phenylalanine ammonia-lyase(PAL)activities while inhibiting vitamin C degradation and weight loss.Furthermore,it restrained polyphenol oxidase(PPO)activity,ethylene production,and respiration rate increase.These findings demonstrate that 1-MCP not only delays the onset of peel greasiness but also preserves the overall storage quality of‘Yuluxiang’pear at a temperature of 20℃.This study presents a novel approach for developing new preservatives to inhibit pear fruit peel greasiness and provides a theoretical foundation for further research on pear fruit preservation.

Effect of Internal Ethylene and 1-Methylcyclopropene on Ethylene Metabolism and Membrane Lipid Peroxidation in Papaya Fruit

[Objective] This study was conducted to evaluate the effect of 1-methylcy- clopropene （1-MCP） on fruit firmness, and the activity of the enzymes involved in ethylene metabolism and membrane lipid peroxidation. [Method] The nearly ripe fruits of the papaya cultivar Risheng were randomly assigned to one of four groups. Two groups were treated under hypobaric and hypoxic （HH） atmosphere condition for six hours, and immediately soaked in deionized water （HH alone）, or fumigated with 2.0 mg/L 1-MCP （HH＋I-MCP） for 24 h. The other two groups untreated under HH condition were also soaked in deionized water （negative control）, or fumigated with 2.0 mg/L 1-MCP （1-MCP alone） for 24 h. After that, the fruits of all the four treatments were stored at room temperature （23＋1） ℃. Cell membrane permeability, fruit firmness, respiration rate, ethylene release rate, SOD activity, POD activity, CAT activity, MAD content and LOX activity were measured once every three days during storage. [Result] Treatment with 1-MCP delayed the occurrence of the peaks of respiration rate and ethylene release rate, significantly reduced the accumulation of malondialdehyde （MDA）, and inhibited the decrease in papaya fruit firmness. Compared with the control, 1-MCP treatment significantly increased the SOD （su- peroxide dismutase）, POD （peroxidase） and CAT （catalase） activity, reduced the ac- tivity of lipoxygenase （LOX）, a product of lipid peroxidatlon in membranes, and in- hibited ethylene biosynthesis, thus delaying the aging process and prolonging the storage life of papaya fruits. [Conclusion] The results will provide a theoretical basis for analvzina the key factors controllinq postharvest maturity and aging of papaya fruits.

Mechanism and Kinetics of Thermal Dissociation of Inclusion Complex of β-Cyclodextrin and 1-Methylcyclopropene

The inclusion-complex of CD-MCP （β-cyclodextrin （β-CD） including 1-methylcyclopropene （1-MCP）） was prepared and characterized. Basing on programmed-heating procedure and weight-temperature analysis, as well as the application of Satava-Sestak＇s, Ozawa＇s and Kissinger＇s methods, the mechanism and kinetics of thermal dissociation of this inclusion complex were studied. An additional mass loss is found at 170-180℃. The mechanism of thermal dissociation of CD-MCP is dominated by a one-dimensional random nucleation and subsequent growth process （A2/3）. The activation energy Es and the pre-exponential factor AS for the process are 102.14 kJ/mol and 3.63×10^10s^-1, respectively. This ES value shows that there is no strong chemical intere, ctions between β-CD and 1-MC;P,

Preharvest application of 1-methylcyclopropene and Ethephon altered cuticular wax biosynthesis and fruit quality of apples at harvest and during cold storage

To investigate the roles of Harvista(a sprayable 1-methylcyclopropene,1-MCP,available component is 150 g·hm^(-2))and Ethephon(1 mL·L^(-1))applied at preharvest in apple(Malus domestica Borkh.'Golden Delicious')fruit cuticular wax biosynthesis,the expression of genes related to fruit cuticular wax biosynthesis and ethylene biosynthesis and signaling,ethylene production rate,respiration rate,wax constituents and structure,and fruit quality were determined at harvest and during cold storage.The results showed that 1-MCP inhibited the expression levels of ethylene biosynthesis and signaling-related genes,decreased fruit ethylene production,and inhibited the expression of Md LACS1,Md CER6,Md CER4 and Md WSD1,which resulted in decreases in alcohols,acids,and esters content in fruit cuticular wax.1-MCP also reduced fruit dropping rate from 17.17%to 12%;maintained fruit firmness,soluble solids,titratable acidity during cold storage;showed about as one fifth in the total length and one third in the widest width of wax crack as that in control at harvest.In contrast,Ethephon produced the opposite effects.In conclusion,1-MCP inhibited fruit ethylene biosynthesis and signaling,and influenced fruit cuticular wax biosynthesis.Thus,the fruit cuticular wax constituents and structure was altered,and the fruit quality were maintained at harvest and during cold storage.The results provide a new technology for improving apple fruit harvest and postharvest quality by preharvest application of sprayable 1-MCP.

Current progress in application of 1-Methylcyclopropene to improve postharvest quality of cut flowers

Floral senescence or petal abscission that determine cut flower quality and longevity are primarily caused by the regulation of endogenous ethylene and perception of endogenous and exogenous ethylene in floral organs(such as the petals and gynoecium),in ethylene-sensitive species.Several attempts have been made to prolong flower quality and longevity using silver ions and other chemicals that inhibit ethylene production and perception in floral organs.Among the available compounds,1-methylcyclopropene(1-MCP)has been increasingly used as ethylene-action/perception inhibitor because of its ability to inhibit ethylene action/perception and it is environmentally safe.Although 1-MCP has been used in several cut flower crops for about 25 years,there has been no review yet that critically emphasizes the benefits or difficulties of using this compound for the prolongation of cut flower quality and longevity.Here,we outline the role of 1-MCP in inhibiting ethylene action/perception and petal senescence in cut flowers through the blocking of receptor binding sites,and we discuss how various factors affecting efficacy of 1-MCP(such as concentration,treatment time and temperature,genotype,and flower stage)are involved in the achievement of flower longevity.Moreover,we highlight the advantages of applying nonvolatile and liquid 1-MCP formulation types,as opposed to using the conventional 1-MCP treatment(powder formulation type).We expect that this review will provide useful information for the future utilization of 1-MCP for the maintenance of cut flower longevity.

High Temperature and the Ethylene Antagonist 1-Methylcyclopropene Alter Ethylene Evolution Patterns,Antioxidant Responses,and Boll Growth in Gossypium hirsutum

The cotton (Gossypium hirsutum L.) crop experiences high temperatures during flowering and boll development, but information regarding the impact of ethylene inhibition and high temperature on early boll development is limited. The objective of this study was to determine the effects of high temperature and the anti-ethylene compound 1-methylcy-cloprone (1-MCP) on G. hirsutum boll development. Treatments consisted of temperature regime (38/20?C and 30/ 20?C), 1-MCP treatment, and days past anthesis (DPA). High temperature decreased ethylene synthesis by 61% at 2 DPA, and 1-MCP caused a 40% decrease in ethylene production at 1 DPA. Glutathione reductase activity increased under high temperature, whereas superoxide dismutase activity (SOD) and membrane peroxidation (malondialdehyde content) remained unchanged. 1-MCP treatment did not affect GR activity in developing bolls. High temperature and 1-MCP treatment increased the weight of cotton bolls collected 8 DPA with an increase of 0.7 and 1 g, respectively. We propose that increased GR activity in bolls exposed to high temperature may mitigate oxidative damage. Additionally, we conclude that ethylene inhibition (either high temperature or 1-MCP-induced) immediately after flowering (1 or 2 DPA) could potentially have positive impacts on early boll growth.

Effects of Exogenous Ethylene and 1-Methylcyclopropene on of DpXTH Gene Expression in Dahlia(Dahlia pinnata Cav.) Flowers

Objectives] This study was conducted to investigate the regulatory mechanism of ethylene on DpXTH gene expression in dahlia （ Dahlia pinnata Cav.） flower petals.[Methods] Dahlia flower petals were treated with ethephon （ETH） and 1-methylcyclopropene （1-MCP） at different flower development stages and for different durations in vivo . Then, the relative expression of DpXTH1 and DpXTH2 in flower samples in different treatments was quantified with real-time PCR, using β-actin as a reference gene.[Results] The expression of DpXTH1 and DpXTH2 in ETH- or 1-MCP-treated dahlia flower petals increased at first, and decreased subsequently with increasing duration of treatment. The relative expression of both DpXTH1 and DpXTH2 in ETH-treated samples reached the maximum level at 12 h. Moreover, the relative expression of both DpXTH1 and DpXTH2 in dahlia flower petals treated with ETH at early stage of flowering was higher than that of the samples treated at other flowering stages; in 1-MCP-treated samples, it was higher at full-flowering stage than at other stages. The maximum relative expression levels of DpXTH1 and DpXTH2 in ETH- and 1-MCP-treated samples were all higher than those of control.[Conclusions] Exogenous ethylene can accelerate the opening of dahlia flowers, and 1-MCP can inhibit this process, so they can be used to regulate development of dahlia flowers.

Effect of Internal Ethylene and 1-methylcyclopropene on Ripening of Papaya Fruit(Carica papaya L.)

With papaya fruit （ Carica papaya ） as an experimental material, variations of its physiological indices were investigated through hypobaric hypoxia （HH, 10 kPa, 1.5 kPa O 2） treatment followed by fumigation with 1-methylcyclopropene （1-MCP）, so as to discuss the effect of interaction between 1-MCP and internal ethylene on ripening of papaya fruit during postharvest storage. The results indicated that HH treatment significantly reduced internal ethylene concentration （IEC） by 57.6%. However, there were no remarkable differences in other physiological indices between various treatments and control （CK）. The 1-MCP and HH＋1-MCP treatments could significant postpone the softening process and color change of papaya fruit. The two treatments achieved the effects of significantly inhibiting the occurrence of fruit disease and the increase of pericarp cell membrane permeability, effectively delaying the variation of soluble solid and Vc contents, and better maintaining soluble solid and Vc contents in fruit, thereby better keeping nutritional substances and flavor quality in fruit in later storage period and prolonging the storage time of fruit.

Postharvest Application of 1-Methylcyclopropene Maintains Quality and Extends the Shelf-Life of Zizania latifolia during Storage at 25˚C

The effects of 1-methylcyclopropene (1-MCP) on postharvest quality of Zizania latifolia during storage at 25˚C were investigated. The results pointed out that a postharvest application of 1-MCP maintained the good visual appearance of fresh Z. latifolia, inhibited browning, mildew and weight loss at the bottom of Z. latifolia, and there is no significant changes on L, a*, b* and ΔE during the whole storage period. In addition, 1-MCP treatment inhibited the respiratory intensity of Z. latifolia during the first three days of storage, but it was significantly higher than that of the control on the sixth day of storage. Compared with the control, 1-MCP treatment maintained relatively high SOD, CAT, APX activities and low PAL, POD, PLD, lipase and LOX activities, delayed the decline of AsA content, reduced the accumulation of O2-, H2O2 and MDA, and ultimately maintained the integrity of cell structure and delayed the senescence of Z. latifolia. In addition, positive effects of 1-MCP on maintaining the cell structure integrity were observed in this investigation throughout the storage period at 25˚C.

Effects of 1-methylcyclopropene and modified atmosphere packaging on fruit quality and superficial scald in Yali pears during storage

The Yali pear（Pyrus bretschneideri Rehd.） is susceptible to superficial scald during prolonged cold storage and at shelf life. This study investigated the effects of 1-methylcyclopropene（1-MCP） and modified atmosphere packaging（MAP） on changes of fruit quality and superficial scald during cold storage and at shelf life in Yali pear. Compared with MAP, the combination of MAP and 1-MCP（MAP＋1-MCP） treatment reduced the carbon dioxide and ethylene content inside the packaging bag. The 1-MCP, MAP, and MAP＋1-MCP treatments reduced the superficial scald index, malondialdehyde content, O2^-· production rate and relative conductivity and inhibited the accumulation of α-farnesene and conjugated trienes in the peel. 1-MCP and MAP＋1-MCP treatments maintained a higher phenolic content and enhanced the catalase and superoxide dismutase activities in the fruit, while reduced activities of lipoxygenase and polyphenol oxidase in the peel preceding the onset of superficial scald. Comprehensive analysis indicated that the MAP＋1-MCP treatment is the most effective method tested for improving the quality of Yali pears during cold storage and at shelf life.

Effects of 1-Methylcyclopropene on NO Content,NOS Activity,and H_2O_2 Content in Postharvest Suli Pears

Studies were conducted over the effects of 1-methylcycolpropene （1-MCP） treatment on postharvest life of Suli pears （Pyrus bretschneideri Rehd.） stored at room and cold temperatures on nitric oxide （NO）,nitric oxide synthase （NOS） activity,and hydrogen dioxide （H2O2）.Results showed that the 1-MCP treatment had little effect on total soluble solids （TSS） at both room and cold temperatures.1-MCP delayed softening of Suli pear fruits,decreased respiratory rate and H2O2 accumulation,and increased NO and NOS activity at room temperature storage,while the effect of cold temperature storage was relatively inferior.There was a significant positive correlation between NOS activity and NO content.It is concluded that 1-MCP had effects on endogenous NO content and accumulation of H2O2.Similarly,H2O2 acting as a signaling molecule via regulating NO level affects the ripening and senescence of Suli pears.

Regulation of Propylene and 1-Methylcyclopropene on Expressions of ACS and ACO Genes in Persimmon Fruits

The regulation of postharvest treatment with propylene and 1-MCP on ethylene release rate and expressions of 1- aminocyclopropane-l-carboxylic acid synthase （ACS） and 1-aminocyclopropane-1-carboxylic acid oxidase （ACO） genes in Fuping Janshi persimmon （Diospyros kaki L.） fruit were investigated. Fruits were treated with propylene and 1-MCP, then stored at 20℃, ethylene release rate of the treated fruits was measured at regular intervals and RNA was extracted for Northern blotting analysis. The results suggested that treatment with propylene accelerated the expressions of ACS and ACO genes and then enhanced the ethylene biosynthesis, while treatment with 1-MCP inhibited the expressions of two genes and their ethylene biosynthesis. Furthermore, different effects on expressions caused by treatments with propylene and 1-MCP existed in various fruit tissues, the inhibitory effect on ACS and ACO genes by 1-MCP was the strongest in pericarp, followed by pulp and core tissues, in the area near fruit stalk, the inhibitory effect was the weakest. While the enhanced effect on ACS and ACO genes by propylene increased from pulp, core, and pericarp to the area near fruit stalk. Expression of each member of ACS and ACO families in various tissues was also completely different, in control and propylene treatment, DKACS3 gene just expressed in the area near fruit stalk and did not express in other tissues, but DKACS2 gene expressed in four tissues by treatment with propylene.

Influences of 1-methylcyclopropene-containing papers on the metabolisms of membrane lipids in Anxi persimmons during storage

Objectives:The aim of this work was to analyse the effects of 1-methylcyclopropene(1-MCP)treatment on the metabolisms of membrane lipids in postharvest Anxi persimmons during storage.Materials and methods:Anxi persimmon(Diospyros kaki L.f.cv.Anxi)fruits were treated by paper containing 1-MCP with a concentration of 1.35μl/l.The cellular membrane permeability was analysed by the electric conductivity meter.The activities of lipoxygenase(LOX),phospholipase(PLD)and lipase were determined by spectrophotometry.The component and relative amounts of membrane fatty acids were determined using gas chromatograph(GC).Results:The 1-MCP-treated Anxi persimmons manifested a lower electrolyte leakage rate,lower LOX,PLD and lipase activities,higher levels of unsaturated fatty acids(USFAs),higher ratio of USFAs to saturated fatty acids(SFAs)(U/S),higher index of USFAs(IUFA),but lower levels of SFAs.Conclusions:The degradation and the metabolisms of membrane lipids could be suppressed by 1-MCP treatment,which might be accountable for the delaying softening of postharvest Anxi persimmons during storage.

Paper-containing 1-methylcyclopropene treatment suppresses fruit decay of fresh Anxi persimmons by enhancing disease resistance

Objectives:The purpose of this work was to evaluate the potential application of papers containing 1-methylcyclopropene(1-MCP)postharvest treatment for suppressing fruit decay of fresh Anxi persimmons and its possible mechanism.Materials and methods:Anxi persimmon fruit were treated with papers containing 1-MCP at the dosage of 1.35μL/L and stored at(25±1)℃and 85 per cent relative humidity for 35 days.During storage,the fruit decay rate and lignin content were evaluated,and the content of total phenolics,the activities of phenylalanine ammonia lyase(PAL),polyphenol oxidase(PPO),peroxidase(POD),chitinase(CHI),and p-1,3-glucanase(GLU)were determined by spectrophotometry.Results:The 1-MCP-treated persimmons displayed a lower fruit decay rate,but higher contents of lignin and total phenolics,higher activities of PAL,PPO,POD,CHI,and GLU.Conclusions:The treatment with 1-MCP could inhibit the fruit decay of postharvest Anxi persimmons,which might be because 1-MCP enhanced fruit disease resistance by increasing the activities of disease resistance-associated enzymes and retaining higher contents of disease resistance-related substances in postharvest fresh Anxi persimmons.These findings indicate that papers containing 1-MCP at the dosage of 1.35μL/L have potential application in suppressing fruit decay and extending storage life of postharvest fresh Anxi persimmons.

Integrative metabolomics and transcriptomics analyses reveal pivotal regulatory mechanisms of 1-methylcyclopropene in maintaining postharvest storage quality of‘Fuji’apples

In this study,integrative metabolomics and transcriptomics analyses were conducted to investigate the effects of 1-methylcyclopropene(1-MCP)on apple fruit quality during long-term cold storage.The results showed that 1-MCP(1μL/L)treatment could maintain fruits apparent quality(i.e.external color and firmness),inhibit the increase of rot rate and soluble solids content/titratable acidity ratio,decrease ethylene release,and respiratory intensity during cold storage,and extend shelf life.Moreover,1-MCP had long-term effects on the accumulation of many qualities related to metabolite and gene expression in fruits.1-MCP affected genes related to metabolism at the early stage of storage,specifically those of the glycolysis and tricarboxylic acid cycle pathways.Genes related to the degradation of sucrose,starch,and cellulose were inhibited,and some starch and cellulose synthesis genes were up-regulated by 1-MCP.Apart from ethylene synthesis and signal transduction being inhibited by 1-MCP,several enzymes(pectinesterase,pectate lyase,polygalacturonase)were involved in pectin degradation,and degradation products of the cell wall(i.e.D-galacturonic acid and D-glucuronic acid)were also strongly inhibited,further maintaining fruit firmness.Cysteine,as precursor glutathione(GSH)related to plant resistance,up-regulated the synthase gene.However,the expression of genes related to cyanoalanine syn-thase and amino acid utilization pathways was suppressed by 1-MCP Collectively,1-MCP could maintain the postharvest quality of apple fruits.

Effects of 1-methylcyclopropene treatment on the quality of red‘Fuji’apples fruit during short-term storage

Objectives:The aim of this study is to determine the mechanism through which 1-methylcyclopropene(1-MCP)affects the quality of red'Fuji'apples,which were stored for a short duration.Materials and Methods:Red‘Fuji’apples were treated with 1-MCP(1.0μl/L),stored at 25℃for0,12,24,48,72,and 96 h,and ethylene pro-duction was measured.An integrated metabolomic and transcriptomic analysis was performed on apples stored for 24 h.Results:The release of ethylene was significantly delayed from red‘Fuji’apples subjected to 1-MCP treatment.By performing an integrated transcriptome and metabolome analyses,we identified 117 differentially expressed genes(DEGs)and 44 differentially accumulated metabol-ites(DAMs).By performing functional enrichment analysis,we found that DEGs were involved in the following pathways;carbon metabolism(LPD2,gpmA,LTA2,ACC,PSAT1,MdCAS2),phytohormone signal transduction(EBF 7),amino acid metabolism(MdACS-1),fatty acid metab-olism(LOX1.5,KCS4,KAST),energy metabolism(Lhcb1,Lhcb6,PsbY,GPDHC1,PUMP5I,metabolic pathways(TRE1,HEXO1)and cell wall metabolism(CSLG2).Thus,these DEGs were involved in the ripening of fruits,and they controlled the quality of fruits at the post-harvest stage.The metabolites were enriched with DAMs.These were found to be individually involved in the metabolic pathway,secondary metabolite bio-synthesis,phenylpropanoid biosynthesis,flavonoids,and flavonol synthesis.Conclusions:The results indicate that 1-MCP inhibits the biosynthesis of ethylene and suppresses energy metabolism.Moreover,it also downregulates metabolic pathways and the enzymatic genes related to fruit quality.Therefore,1-MCP delays the ripening of fruits at the post-harvest stage.This study helps us understand how 1-MCP treatment affects the ripening and quality of fruits.

骨形成肽1和聚多巴胺复合涂层修饰提高聚醚醚酮表面活性

背景:聚醚醚酮具有与人体皮质骨相近的弹性模量及良好的射线透射性、化学稳定性和生物相容性等优点,有望应用于口腔种植领域,然而其具有生物惰性,难以与周围组织形成骨结合,因此如何提高聚醚醚酮的表面活性是目前的主要问题。目的:分析聚醚醚酮表面骨形成肽1和聚多巴胺复合涂层的促成骨和成血管作用。方法:将聚醚醚酮片浸泡于多巴胺溶液中24 h,制备聚醚醚酮-聚多巴胺材料;将聚醚醚酮-聚多巴胺材料浸泡于骨形成肽1溶液中24 h,制备聚醚醚酮-聚多巴胺-骨形成肽1材料,表征材料的微观形貌、亲水性与元素组成。将骨髓间充质干细胞分别接种于聚醚醚酮、聚醚醚酮-聚多巴胺、聚醚醚酮-聚多巴胺-骨形成肽1材料表面,通过活死细胞染色和细胞骨架染色评估细胞活性与黏附状态,茜素红和骨钙素免疫荧光染色检测细胞成骨分化能力。将人脐静脉内皮细胞分别接种于3组材料表面,通过活死细胞染色和细胞骨架/血管内皮生长因子免疫荧光染色评估细胞活性及成血管水平。结果与结论:①扫描电镜下可见聚醚醚酮材料表面光滑,聚醚醚酮-聚多巴胺材料表面出现凹凸不平的沉积物,聚醚醚酮-聚多巴胺-骨形成肽1材料表面有小颗粒突起;接触角测试结果显示,聚醚醚酮-聚多巴胺-骨形成肽1材料的亲水性优于其他两种材料;X射线光电子能谱测试结果显示,骨形成肽1成功修饰于聚醚醚酮材料表面;②活死细胞染色和细胞骨架染色显示,相较于其他两种材料,聚醚醚酮-聚多巴胺-骨形成肽1材料可提高骨髓间充质干细胞的活性与黏附;茜素红和骨钙素免疫荧光染色显示,相较于其他两种材料,聚醚醚酮-聚多巴胺-骨形成肽1材料可促进骨髓间充质干细胞的成骨分化;③活死细胞染色和免疫荧光染色显示,相较于其他两种材料,聚醚醚酮-聚多巴胺-骨形成肽1材料可提高人脐静脉内皮细胞的活性与黏附及血管内皮生长因子蛋白的表达;④结果表明,骨形成肽1和聚多巴胺复合涂层可提高聚醚醚酮表面的促成骨和成血管活性。

蠲痹汤含药血清调控线粒体自噬抑制白细胞介素1β诱导的关节软骨细胞损伤

背景:软骨细胞线粒体自噬的缺陷会引起细胞凋亡和基质消失等软骨细胞退行性病变的变化。目的:探讨蠲痹汤含药血清对白细胞介素1β诱导的大鼠膝关节软骨细胞炎症反应和凋亡的影响及可能作用机制。方法:50只雄性SD大鼠随机给予生理盐水、蠲痹汤低、中、高剂量(1.24,2.48,4.96 g/kg)、塞来昔布(阳性药物),连续灌胃2周后获得含药血清。①分离软骨细胞,将其随机分为对照组、白细胞介素1β组、蠲痹汤低、中、高剂量含药血清组及阳性药物血清组。CCK-8法检测细胞存活率、免疫荧光双染检测线粒体自噬水平、免疫荧光检测磷酸化腺苷酸激活蛋白激酶水平、Western blot检测PTEN诱导激酶1/Parkin通路相关蛋白和裂解的半胱氨酸蛋白酶蛋白3表达、ELISA检测炎症因子水平;②分别采用PTEN诱导激酶1 siRNA和Compound C进行干预,探究AMPK/PTEN诱导激酶1/Parkin通路在蠲痹汤含药血清调控线粒体自噬中的作用。结果与结论:①与对照组比较,白细胞介素1β组软骨细胞存活率、Ⅱ型胶原蛋白表达、磷酸化腺苷酸激活蛋白激酶、PTEN诱导激酶1、Parkin和微管相关蛋白1轻链3蛋白水平以及线粒体自噬水平明显降低(P<0.05),而裂解的半胱氨酸蛋白酶蛋白3蛋白水平、白细胞介素6、白细胞介素8和肿瘤坏死因子α水平显著升高(P<0.05);与白细胞介素1β组比较,蠲痹汤各剂量含药血清组和阳性药物血清组上述各项指标呈现相反的变化(P<0.05);②PTEN诱导激酶1 siRNA可显著抑制蠲痹汤含药血清对白细胞介素1β处理软骨细胞线粒体自噬的影响,降低蠲痹汤含药血清对白细胞介素1β诱导的软骨细胞炎症与凋亡的保护作用;Compound C逆转了蠲痹汤含药血清对白细胞介素1β处理软骨细胞中PTEN诱导激酶1/Parkin信号通路的影响。结论:蠲痹汤含药血清通过影响线粒体自噬水平来抑制软骨细胞炎症和凋亡,从而减轻白细胞介素1β诱导的软骨细胞退化,其机制可能与调控AMPK/PTEN诱导激酶1/Parkin通路有关。

组蛋白脱乙酰酶1基因抑制人脐静脉内皮细胞焦亡并减轻动脉粥样硬化及炎性反应

背景:细胞焦亡作为炎症细胞死亡的一种独特形式,在动脉粥样硬化病变的不稳定性中发挥重要作用。目的:探究重组B细胞淋巴瘤2相关蛋白A1(B-cell lymphoma 2-related protein A1,BCL2A1)在动脉粥样硬化中的作用机制。方法:①使用200μg/mL氧化低密度脂蛋白处理人脐静脉内皮细胞24 h以诱导内皮损伤。随后,分别使用50 nmol/L BCL2A1干扰质粒(sh-BCL2A1)和1.5μg/mL组蛋白脱乙酰酶1基因(histone deacetylase 1 gene,HDAC1)过表达载体(pcDNA-HDAC1)转染人脐静脉内皮细胞,或同时转染pcDNA-HDAC1和BCL2A1过表达载体(pcDNA-BCL2A1)。转染后培养48 h检测BCL2A1和HDAC1的表达水平、细胞活力、细胞焦亡水平以及BCL2A1乙酰化水平。②通过高脂喂养APOE-/-小鼠构建动脉粥样硬化小鼠模型进行体内验证。将500μL BCL2A1和HDAC1慢病毒过表达载体分别或同时尾静脉注射到小鼠体内,检测BCL2A1和HDAC1的表达水平以及小鼠动脉组织损伤情况。结果与结论:氧化低密度脂蛋白诱导的人脐静脉内皮细胞中BCL2A1上调,干扰BCL2A1可改善细胞活力并抑制细胞焦亡和炎症反应。此外,氧化低密度脂蛋白诱导的人脐静脉内皮细胞中HDAC1下调,通过促进BCL2A1去乙酰化提高了细胞活力及抑制了细胞焦亡和炎症反应。体内实验表明,BCL2A1在高脂喂养的小鼠动脉组织中高表达,而HDAC1低表达。此外,HDAC1通过促进BCL2A1去乙酰化减轻了高脂喂养诱导的ApoE-/-小鼠动脉组织病变。结果表明,HDAC1可能通过BCL2A1去乙酰化来抑制人脐静脉内皮细胞焦亡进而减轻动脉粥样硬化和炎症反应。

创伤性脊髓损伤急性期前列腺素E1对血管相关因子的调节和微循环功能的保护

背景:前列腺素E1被证明在血管扩张、炎症、白细胞迁移和黏附中发挥调节作用,但其对创伤性脊髓损伤后脊髓微循环的作用尚缺乏深入的研究。目的:探讨在大鼠创伤性脊髓损伤急性期给予前列腺素E1对血管相关因子的调节和微循环功能的保护作用机制。方法:将72只雌性SD大鼠随机分为3组(n=24),即假手术组、脊髓损伤组、前列腺素E1组。后两组用Allen’s打击法建立脊髓损伤的体内模型,前列腺素E1组大鼠在脊髓损伤后15 min内立即尾静脉注射脂质前列腺素E110μg/kg。分别在损伤后2,24 h测定脊髓微循环血流量和血氧饱和度、脊髓微血管直径和面积、脊髓含水量、血管功能调节因子(血浆血管性血友病因子、血栓素A2、前列环素、内皮素1)和炎症因子(肿瘤坏死因子α、白细胞介素1β)的表达。结果与结论:①脊髓损伤后2 h,前列腺素E1组大鼠的脊髓微血管直径及面积、脊髓微循环血流量和血氧饱和度均高于脊髓损伤组(P<0.05),脊髓含水量低于脊髓损伤组(P<0.05),血浆血管性血友病因子、脊髓组织血栓素A2/前列环素及内皮素1质量浓度均低于脊髓损伤组(P<0.05);②脊髓损伤后24 h,前列腺素E1组大鼠的脊髓微血管面积、血流量和血氧饱和度均高于脊髓损伤组(P<0.05),脊髓含水量低于脊髓损伤组(P<0.05),血浆血管性血友病因子、脊髓组织血栓素A2/前列环素及内皮素1、肿瘤坏死因子α、白细胞介素1β的质量浓度均低于脊髓损伤组(P<0.05);③脊髓损伤组大鼠损伤后24 h的脊髓微血管直径及面积、脊髓微循环血流量和血氧饱和度均高于损伤后2 h(P<0.05),血浆血管性血友病因子、脊髓组织血栓素A2/前列环素、肿瘤坏死因子α、白细胞介素1β的质量浓度均高于损伤后2 h(P<0.05),但是脊髓组织内皮素1质量浓度低于损伤后2 h(P<0.05);④前列腺素E1组大鼠损伤后24 h的脊髓微循环血流量和血氧饱和度低于损伤后2 h(P<0.05),脊髓微血管直径及面积、脊髓含水量高于损伤后2 h(P<0.05);⑤以上结果表明,脊髓损伤大鼠伤后即刻静脉给予前列腺素E1,可调节血管功能调节因子、炎症因子并改善脊髓损伤后脊髓微循环,这为寻找治疗急性脊髓损伤的药物提供了潜在的基础。