Polysaccharide extract from Rosa laevigata fruit attenuates inflammatory obesity by targeting redox balance and gut interface in high-fat diet-fed rats

Low-molecular-weight polysaccharides(RLPs)extracted from Rosa laevigata fruits showed multiple biofunctions in Asia.This study aimed to investigate its anti-inflammatory obesity effect in high-fat dietfed rats and further elucidated the underlying molecular mechanism by multi-omics methods.The results showed that RLPs administration had significantly restored immune organ indexes and reduced body weight gain.RNA-seq revealed that the effect of RLPs was partially attributed to its regulation on PPARs signaling by increasing the expressions of Scd,Acox3 and Hmgcs2,and on other redox-related pathways by decreasing the expressions of Cyp2 e1,Il1-r1 and Lbp.Moreover,16 S rRNA sequencing coupled with metagenome sequencing showed that RLPs had significantly reduced the ratio of Firmicutes/Bacteroidetes from 8.01 to2.37,and significantly increased the relative abundances of Alistipes,Prevotella,and Akkermansia from 0.36%,1.10%and 2.61%to 0.65%,2.37%and 4.42%,respectively.Spearman correlation analysis result indicated that the abundances of Lachnospiraceae,Prevotella and Bacteroidales were significantly negatively correlated with obesity phenotype,liver function and inflammatory factors.These results revealed that RLPs exerted significant anti-inflammatory obesity property partially via regulation on gut microbiota interface and the redox balance.Therefore,RLPs could be a promising functional food resource with the potential for redox imb alance-related diseases chemoprevention.

Exploring the Protective Effect of the Ethanolic Extract of Rosa laevigata Michx.Fruit on Rats with Mesangial Proliferative Glomerulonephritis Based on NLRP3 Inflammasome Pathway

Objective:To investigate the effect of the ethanolic extract of Rosa laevigata Michx.fruit on rats with mesangial proliferative glomerulonephritis based on the NLRP3 inflammasome pathway.Methods:Thirty Wistar rats were divided into three groups,a blank control group,a diabetic nephropathy(DN)model group,and an ethanolic extract intervention group,according to the random number table method,with 10 rats in each group.One day before the experiment,basic feeding was initiated for all the rats;the changes in activity and weight of each group of rats were observed and recorded after 7 d,and a rat model of renal function injury was established after 1 d.Results:Compared with the control group,the model group had significantly higher kidney/body ratio,24 h urine protein,serum creatinine(SCr),blood urea nitrogen(BUN),glomerular mesangial cell(GMC)count,and extracellular matrix(ECM)positive area ratio(P<0.05);the same indicators were significantly lower in the intervention group than in the model group(P<0.05).The NLRP3 inflammasome pathway in renal intrinsic cells was activated in the intervention group.The overactivation of NLRP3 inflammasome is known to promote interleukin(IL)-1βrelease,which was inhibited in the intervention group.Conclusion:The ethanolic extract of Rosa laevigata Michx.fruit has a protective effect on renal intrinsic cells and may be related to NLRP3 inflammasome pathway,suggesting that the fruit of Rosa laevigata Michx.has a potential role in protecting renal intrinsic cells from inflammatory damage.NLRP3 inflammasomes are involved in the development of various chronic inflammatory diseases,such as acute and chronic glomerulonephritis and renal fibrosis.

湿地植物燕子花(Iris laevigata)对模拟施氮的响应研究

选取吉林龙湾自然保护区一种常见的湿地植物——燕子花(Iris laevigata Fisch.)为研究对象,通过模拟控制实验研究了其在不同氮素添加处理下形态、生理和繁殖三方面特征的响应规律.研究共设置3个氮素处理梯度,即对照(0)、低氮(5g/m2)和高氮(15g/m2).结果表明:随着氮素添加量的增加,燕子花株高、生物量呈现下降的趋势,对照组的株高和生物量均显著高于其他处理下的株高和生物量;而叶片数各处理间无显著差异(P>0.05).随着氮素添加量的增加,净光合作用速率和叶绿素含量也逐渐下降;而丙二醛、可溶性糖、脯氨酸等呈现增加的趋势.各处理间,植株的开花物候和花部综合特征有所差异,但开花持续时间、花粉数、胚珠数、花粉活力、柱头可授性均无显著差异(P>0.05).燕子花在氮元素增加的情况下,其生长表现并不佳,反而可能受到抑制.在今后湿地的保护管理中,应科学控制湿地周围农田耕作等人为活动过程中氮肥的使用,避免氮素通过地下途径进入湿地对燕子花种群更新和生长产生影响.

The antioxidant activity and hypolipidemic activity of the total flavonoids from the fruit of Rosa laevigata Michx

In the present work, the antioxidant activity in vitro and hypolipidemic activity of the total fla-vonoids (TFs) from the Rosa laevigata Michx fruit was evaluated, and the antioxidant effect in vivo was also discussed. The TFs exhibited a high scavenging effect on 2, 2-diphenyl-1- picrylhydrazyl (?DPPH) with IC50 values 0.01 mg/mL, and a stong reduce power in the test. Hyperli-pemic mice were intragastric administrated with TFs (25, 50 mg/kg/day) for 4 weeks, and fenofi-brate was used as the positive reference sub-stance. After the experiment, the levels of TC (total cholesterol), TG (triglyceride), LDL- C (low density lipoprotein-cholesterol) of the mice ad-ministrated with high-dose of TFs were mark-edly declined by 45.02%, 33.86% and 73.68%, re-spectively, while HDL-C (high density lipopro-tein-cholesterol) was significantly increased com-pared with model group. To investigate the hepatoprotective effect, histopathological assay, ALT (alanine aminotransferase), AST (aspartate aminotransefrase) and ALP (alkaline phosphatase) were also studied, and the results showed that TFs exhibited a favorable effect on liver protec-tion, of which the levels of ALT, AST and ALP were elevated by 55.85%, 29.15% and 25.68%, respectively. Furthermore, the TFs could sig-nificantly decrease the MDA (malondialdehyde) level and improve the levels of CAT (Catalas), SOD (superoxide dismutase), GSH (reduced glutathione), and GPX (glutathione peroxidase) compared with hyperlipemia mice. Our results suggested that TFs has a high antioxidant ac-tivity and hypolipidemic activity, which can be used as a potential medicine for cardiovascular diseases.

RAPD Analysis of Rosa laevigata Michx. from Different Origins

[ Objective] This study aimed to investigate the genetic diversity and phylogenetic relationship of 30 populations of Rosa laevigata Michx. from different origins. [ Method] Using RAPD molecular marker technique, eight effective primers were screened from 35 RAPD random primers for amplification. UPGMA clus- ter analysis was performed using NTSYSpc. ver. 2.02 software. [ Result] A total of 86 loci were amplified using the screened primers, including 84 polymorphic loci, indicating an average polymorphic percentage （PPB） of 97.67%. The similarity coefficients of 30 populations of R. laevigata Michx. ranged from O. 11 to O. 58, and these populations were clustered into groups A, B and C. [ Conclusion ] R. laevigata Michx. exhibits multiple genetic polymorphism. R. laevigata Michx. populations from Guizhou Province have closer genetic relationships.

Study on Thin Layer Drying of <i>Rosa laevigata</i>Michx

The drying process of traditional Chinese medicine Rosa laevigata Michx was investigated in a laboratory scale dryer. Drying experiments were conducted at five temperatures of 40℃, 50℃, 60℃, 70℃ and 80℃. The experimental data were fitted to ten different thin layer drying models to select a suitable model for describing the drying process of Rosa laevigata Michx fruits. The results showed that the Midilli model had the highest value of R2 (0.999767), the lowest χ2 (0.000034) and RMSE (0.003616) among the ten models considered. The drying process of Rosa laevigata Michx fruits consisted of 1st falling rate period and 2nd falling rate period. The effective moisture diffusivity was found in the range of 1.49 - 30.20 × 10-11 m2·s-1. The value of activation energy obtained by plotting lnD versus 1/T is about 36 kJ·mol-1.

Use of Prosopis laevigata Seed Gum and Opuntia ficus-indica Mucilage for the Treatment of Municipal Wastewaters by Coagulation-Flocculation

Prosopis laevigata and Opuntia ficus-indica grow in arid and semiarid regions of Mexico and other countries. Both produce biopolymers with interesting characteristics from the rheological point of view as well as because of their coagulating-flocculating capabilities. Prosopis produce galactomannans inside the endosperm, very similar to those found in guar, locust bean, and tara gums. Opuntia sp. produces mucilage that contains polygalacturonic acid and five neutral sugars. Prosopis seed gum has not been proposed to be used as coagulant-flocculant before. In the case of Opuntia mucilage, some authors have suggested its use in the treatment of waters, using either the mucilage or the whole cladode powder. The use of these products in the treatment of municipal or even industrial wastewaters could give rise to diverse benefits. From the environmental point of view, treated waters with neither Fe nor Al, nor synthetic polymers would be obtained (with less toxicity risk). Besides, the produced sludges would be smaller in amount, with better biodegradability, and lower metals content. From the economical point of view, the use of these biopolymers would give an added value to the Opuntia and Prosopis culture in Mexico, helping small communities to enhance their incomes by producing environmental-friendly products. This work shows that both Prosopis galactomannan and Opuntia mucilage can be used to treat municipal wastewaters with an initial organic charge of about 827 mg/L as COD by the coagulation-flocculation process, with COD removals for the mesquite seed gum of up to 90% (pH 10, dose of 75 mg/L) and of 60% (pH 7, doses of 50 and 150 mg/L). In the case of mucilage, 65% of the initial COD was removed at pH 10 (dose of 50 mg/L). These figures are very promising for the treatment of wastewaters, with environmental-friendly products.

Differences of Wood Elements of Prosopis laevigata from Two Areas of Northeast Mexico

Anatomical features of wood have a great variation among species as a result of genetic and environmental factors. The anatomical heartwood characteristics of Prosopis laevigata species from two areas with differences on temperature and rain precipitation on northeast Mexico were compared. Fibers length (μm), diameter of vessels (μm), and the area of the vessels (μm-2) were measured using light microscopy coupled with a digitized-image analysis system. The differences were statistically analyzed with analysis of variance. Statistical differences between fibers length (p 0.0001) and diameter of the vessels (p 0.001) from the two localities were found. The locality Linares, Nuevo León, Mexico, with higher precipitation and lower temperature, showed higher fiber length and higher diameter of the vessels than China, Nuevo León. Hard environmental conditions, where low precipitation values and high temperatures prevail, condition P. laevigata trees do reduce the risk of losing water.

金樱子叶衍生碳点的制备及其对金樱子不定芽的影响

为了探讨碳点对植物组织培养的影响,本论文以金樱子叶为碳源,采用一步水热法制备荧光碳点(JCDs),再以金樱子茎段为外植体建立再生体系,研究J-CDs对金樱子不定芽生长的影响。结果表明,J-CDs生物相溶性好,在pH值2-8范围内保持稳定,适用于植物组织培养。添加J-CDs可以促进金樱子不定芽生长,实验组金樱子不定芽株高比对照组高3-4倍,植株健壮,枝叶伸展,生长良好。当J-CDs浓度为0.9mg·mL^(-1)时,金樱子不定芽叶绿素总含量和可溶性蛋白含量达到最大值,SOD和POD酶活性最大,分别比对照组增加了56.25%和41.82%。本研究利用金樱子生物质制备荧光CDs,促进金樱子不定芽生长,为植物繁育提供了新方法。

金樱子多糖、车前子多糖和冬葵果多糖体内外抗炎效果比较

旨在对比金樱子多糖、冬葵果多糖、车前子多糖的体内外抗炎活性。体外试验用脂多糖(LPS)诱导小鼠RAW264.7细胞构建炎症模型,分别加入金樱子多糖、车前子多糖、冬葵果多糖,以MTT法检测其对RAW264.7细胞活力的影响,Griess法检测LPS诱导的细胞上清液中NO含量,ELISA检测白细胞介素6(IL-6)、白细胞介素1β(IL-1β)和α肿瘤坏死因子(TNF-α)含量,RT-PCR法检测炎症细胞因子TNF-α、IL-6、IL-1β的mRNA表达量;体内试验用葡聚糖硫酸钠(DSS)构建小鼠溃疡性结肠炎模型,同时多糖组和阳性药组的小鼠分别灌服多糖(200 mg/kg)和5-氨基水杨酸(200 mg/kg),空白组和DSS组灌服生理盐水,采用ELISA检测结肠组织中炎症因子TNF-α、IL-6、IL-1β的浓度,以结肠长度、组织病理学评分比较结肠组织病理变化。体外试验结果发现:在同一浓度下,与模型组(LPS)相比,对炎性介质(NO、TNF-α、IL-6、IL-1β)含量和促炎细胞因子(TNF-α、IL-6、IL-1β)的mRNA表达量抑制作用以金樱子多糖组最高,车前子多糖组次之,而冬葵果多糖组抑制作用不明显;体内试验结果发现:与DSS组相比,金樱子多糖组的疾病活动指数(DAI)显著降低(P<0.05),结肠长度极显著增加(P<0.01),组织病理学评分和结肠组织中TNF-α、IL-6、IL-1β水平极显著降低(P<0.001),结肠组织中IL-10水平极显著升高(P<0.001),其次是车前子多糖组的结肠长度显著增加(P<0.05),组织病理学评分和结肠组织中TNF-α、IL-1β水平显著降低(P<0.05),结肠组织中IL-6水平极显著降低以及IL-10水平极显著升高(P<0.01),而冬葵果多糖组的指标变化不明显。试验结果表明,与车前子多糖和冬葵果多糖相比,金樱子多糖在体内外的抗炎效果最佳,可以作为多糖类抗炎剂的候选药。

金樱子果实中蔷薇酸的快速富集方法及活性研究

金樱子果实中富含一种5环三萜类活性成分——蔷薇酸(EA),这是一种重要的天然产物,因其多样的药理活性而被广泛研究。由于传统的分离纯化工艺效率低、耗时长,因此亟需一种方便快捷的方法提高EA的富集效率。通过一种自制的减压真空柱色谱装置对金樱子果实中富含的蔷薇酸进行富集,该方法不仅省时省力,极大提高了EA的富集效率,同时也为其他天然活性成分的快速富集提供了一种真实有效的方法。此外,对EA进行分子对接,发现它与乙酰胆碱酯酶(AChE)活性位点的结合能为-36kJ·mol^(-1),具有高度亲和力;对EA进行乙酰胆碱酯酶活性研究,发现在质量浓度为120μg·mL^(-1)时,EA和阳性药加兰它敏对AChE的抑制活性分别可达83.20%和92.91%,这意味着EA具有抗阿尔兹海默症的潜在活性。

金樱子(Rosa laevigata Michx.)叶片直接再生不定芽体系的建立

以金樱子组培苗的划伤叶片为外植体,研究了不同植物生长调节剂配比组合、暗培养时间、添加物L-脯氨酸、叶片不同部位对不定芽直接再生的影响。结果表明:当TDZ浓度为1.5 mg·L^-1、NAA浓度为0.000 5 mg·L^-1时,不定芽直接发生率最高,为9.5%;不同暗培养时间对不定芽直接诱导具有一定影响,暗培养时间为10 d时不定芽的直接发生率最高,为10.5%;添加了不同浓度的L-脯氨酸后不定芽的诱导率不增反降,所以以不添加L-脯氨酸为宜;叶片不同部位的再生率比较中,仅带叶叶柄可直接诱导出不定芽。综上所述,叶片直接再生不定芽的诱导方法是,以金樱子带叶叶柄为外植体,在直接诱导培养基上1/2MS+TDA 1.5 mg·L^-1+NAA 0.000 5 mg·L^-1+CH 100 mg·L^-1+AgNO3 10 mg·L^-1+蔗糖30 g·L^-1+琼脂7.5 g·L^-1,暗培养10 d后,转至正常光周期下培养3周左右,不定芽诱导率最高,为10.5%。所得不定芽在增殖培养基MS+NAA 0.1 mg·L^-1+6-BA 1.0 mg·L^-1生长3周后,增殖系数可达到3.5左右;将丛生芽切割成单芽后转入不含任何激素的MS培养基壮苗3周,幼苗可长高至3~5 cm;再转入MS+NAA 0.1 mg·L^-1培养基中生根,1个月后生根率达95%。

金樱子总多酚提取工艺优化及抗氧化活性研究

该研究采用超声提取法提取金樱子总多酚,以总多酚得率为评价指标,通过Plackett-Burman(PB)试验设计、单因素试验及Box-Behnken(BB)响应面法优化金樱子总多酚的提取工艺,并对其抗氧化活性进行评价。结果表明,金樱子总多酚最佳超声提取工艺为:液料比21∶1(mL∶g),乙醇体积分数61%,超声时间39 min,提取温度48℃。在此优化条件下,总多酚得率为3.84%,比优化前(2.74%)提高40.15%。金樱子总多酚具有较强的抗氧化活性,金樱子总多酚溶液质量浓度为1.2 mg/mL时,金樱子多酚对1,1-二苯基-2-三硝基苯肼(DPPH)及2,2'-联氨-双(3-乙基苯并噻唑啉-6-磺酸)二胺盐(ABTS+)自由基的半抑制浓度(IC50)值分别为0.502 mg/mL、0.425 mg/mL。

金樱叶的化学成分及抗炎活性研究

目的对金樱子Rosa laevigata Michx.叶的化学成分及其抗炎活性进行研究,为进一步开发其药用潜能提供科学依据。方法用大孔树脂色谱、ODS色谱及高效液相色谱法对金樱叶乙醇提取物进行分离纯化,根据理化性质和波谱数据对所得化合物进行结构鉴定,用荧光素酶活性检测法和ELISA法对部分化合物进行体外抗炎活性检测。结果分离得到9个化合物,分别鉴定为:isopinfaenoic acid-28-O-β-D-glucopyranoside(1);2α,3β,23-三羟基乌苏-12,19-二烯-28-O-β-D-吡喃葡萄糖苷(2);2α,3β,19α-三羟基乌苏-12-烯-28-O-β-D-吡喃葡萄糖苷(3);苦莓苷F1(4);2α,3β,23-三羟基乌苏-12,18-二烯-28-O-β-D-吡喃葡萄糖苷(5);5,7,3′,4′-四羟基-4′′-O-甲基-3-O-β-D-葡萄糖黄酮苷(6);芦丁(7);槲皮苷(8);异落叶松脂素-9-O-β-D-吡喃葡萄糖苷(9)。用脂多糖(LPS)刺激的NF-κB-293和RAW264.7W 2种炎症细胞模型筛选出具有不同程度抗炎活性的化合物1、5、8。结论从蔷薇属植物金樱叶中共分离鉴定了9个化合物,其中化合物2、3、6、9为首次从该属植物中分离得到,化合物1、5、8具有不同程度的抗炎作用。

基于网络药理学与分子对接技术探究金樱子治疗高血压的作用机制

目的:探讨金樱子治疗高血压的有效成分、潜在作用靶点及其机制。方法:通过TCMSP、ETCM、BATMAN-TCM、GeneCards等数据库筛选金樱子的有效成分、作用靶点及高血压相关靶点。采用Cytoscape3.9.1软件构建“疾病-活性成分-靶点”网络。利用STRING数据库构建蛋白质相互作用网络,并通过MCODE插件筛选核心靶点。借助DAVID数据库对药物和疾病共同靶点进行GO富集分析以及KEGG通路富集分析。应用Autodock软件进行分子对接验证。结果:通过网络药理学分析共获得24种有效成分、610个药物靶点和517个疾病靶点以及127个药物和疾病交集靶点。通过筛选共获得包括JUN、TNF、IL6等在内的10种核心靶点以及包括3-氢化松苓酸甲酯、野鸦椿酸等在内的有效成分。通过GO和KEGG富集分析找出了流体剪切应力和动脉粥样硬化、HIF-1信号通路等关于脂质代谢、氧化应激和炎症反应以及心血管保护的通路。结论:金樱子治疗高血压的机制是多成分、多靶点、多通路的。

燕子花WRKY11基因克隆、结构分析及功能验证

为了开发燕子花WRKY家族的功能基因资源,验证WRKY11转录因子与花期调控相关性,更好地服务于燕子花定向分子育种。以东北林业大学苗圃的燕子花(Iris laevigata)为材料,燕子花花被片为cDNA模板,克隆燕子花IlWRKY11基因,对IlWRKY11蛋白质结构进行生物信息学分析、亚细胞定位,并通过该基因转化模式植物拟南芥,开展WRKY11转录因子对花期调控方面的功能解析。结果表明:以燕子花总RNA反转录的cDNA为模板,克隆获得IlWRKY11基因的长度为888 bp,编码295个氨基酸;IlWRKY11蛋白的相对分子质量为32291,分子式为C_(1398)H_(2237)N_(419)O_(421)S_(20),属于不稳定亲水性蛋白,pCAMBIA1300-IlWRKY11-GFP植物过表达载体确定IlWRKY11蛋白定位在细胞核,IlWRKY11蛋白与单子叶植物深圳拟兰(Apostasia shenzhenica)、石刁柏(Asparagus officinalis)的关系较近。过表达IlWRKY11基因的拟南芥抽薹时间和开花时间比对照提前2~3 d。通过对转基因拟南芥和转空载对照的成花期相关基因表达量进行qRT-PCR检测,推测IlWRKY11基因通过调控赤霉素途径相关基因GA20OX和春化途径基因VRN1以及年龄途径基因SPL3参与花期调控,说明IlWRKY11基因(ON399551)对燕子花花期调控具有重要作用。

光果金樱子三萜类化合物的分离鉴定与抗SARS-CoV-2活性评价

为了解光果金樱子(Rosa laevigata var. leiocapus)的三萜类化学成分及其抗严重急性呼吸综合征冠状病毒2活性(severe acute respiratory syndrome coronavirus 2, SARS-CoV-2),运用多种色谱分离技术从其95%乙醇提取物中分离得到15个三萜化合物。根据理化性质及波谱数据,分别鉴定为:laevigaterpeneA(1)、2α,23-二羟基齐墩果酸(2)、1β-羟基蔷薇酸(3)、3β-(p-hydroxytrans-cinnamoyloxy)olean-12-en-28-oic acid (4)、3β-反式对羟基肉桂酰氧基-2α-羟基齐墩果酸(5)、2α,3α-二羟基-12-烯-28-齐墩果酸(6)、坡模酸(7)、桦木酸甲酯(8)、2α,3α,19α,23-四羟基-12-烯-乌苏酸(9)、乙酰基-11α-甲氧基-β-乳香酸(10)、阿江榄仁尼酸(11)、2α,3α,19α-trihydroxy-28-norurs-12-ene (12)、蔷薇酸(13)、2α,19α-dihydroxy-3-oxo-12-ursen-28-oic acid (14)和齐墩果酸(15)。所有化合物均为首次从光果金樱子中分离获得,其中化合物4、10、12为首次从蔷薇属植物中分离得到。化合物8和15对SARS-CoV-2主蛋白酶(mainprotease,简称Mpro)具有较强的抑制作用,IC50值分别为(6.74±0.33)和(5.19±0.25)μmol/L,具有潜在的抗SARS-CoV-2活性。

基于指纹图谱研究金樱子肉饮片、标准汤剂、配方颗粒相关性

目的 建立金樱子肉饮片、标准汤剂、配方颗粒的HPLC指纹图谱,并分析其相关性和差异性。方法 采用HPLC建立金樱子肉饮片、标准汤剂、配方颗粒指纹图谱分析方法,以乙腈为流动相A,以0.1%磷酸溶液为流动相B,梯度洗脱,流速1 mL/min;柱温30℃,检测波长260 nm,利用相似度评价、皮尔逊相关性分析、聚类分析(CA)、主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA)对金樱子肉饮片、标准汤剂及配方颗粒指纹图谱进行评价和分析。结果 金樱子肉饮片、标准汤剂、配方颗粒的HPLC指纹图谱均确定了8个共有峰,并指认了其中的1、6、7、8号峰,分别为没食子酸、鞣花酸-4-O-α-L-阿拉伯糖苷、鞣花酸和槲皮素-3-O-β-D-吡喃葡糖苷酸;三者与相应对照指纹图谱的相似度均大于0.950,三者对照指纹图谱间的相似度均大于0.990;相关性分析的结果表明大部分共有峰成显著相关关系;通过系统聚类分析,表明饮片、标准汤剂、配方颗粒三者之间无显著性差异。主成分分析结果显示,两个主成分的累计方差贡献率为72.13%;OPLS-DA将45个样品分为4类,并筛选出4个VIP值大于1的差异性成分。结论 金樱子肉饮片、标准汤剂、配方颗粒的主要化学成分组成基本一致,可为金樱子肉配方颗粒生产过程的质量控制提供参考。

金樱子总黄酮上调miR-1247-3p表达对缺氧/复氧诱导心肌细胞损伤的影响

目的:观察金樱子总黄酮(RLMTF)对缺氧/复氧(H/R)诱导的心肌细胞损伤的影响及可能作用机制。方法:采用H/R诱导大鼠心肌细胞H9C2建立细胞损伤模型,采用不同剂量的金樱子总黄酮处理细胞,实验分为Con组、H/R组、H/R+RLMTF-L组、H/R+RLMTF-M组、H/R+RLMTF-H组、H/R+miR-NC组、H/R+miR-1247-3p组、H/R+RLMTF+anti-miR-NC组、H/R+RLMTF+anti-miR-1247-3p组。使用试剂盒检测丙二醛(MDA)水平与超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性;流式细胞术检测细胞凋亡率;实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-1247-3p表达。结果:与Con组比较,H/R组MDA水平、细胞凋亡率和Cleaved-Caspase 3、Cleaved-Caspase 9蛋白水平升高(P<0.05),miR-1247-3p表达量和SOD、GSH-Px活性降低(P<0.05)。与H/R组比较,H/R+RLMTF-L组、H/R+RLMTF-M组、H/R+RLMTF-H组MDA水平、细胞凋亡率降低(P<0.05),miR-1247-3p表达量和SOD、GSH-Px活性升高(P<0.05),且呈剂量依赖性。与H/R+miR-NC组比较,H/R+miR-1247-3p组MDA水平、细胞凋亡率降低(P<0.05),SOD、GSH-Px活性升高(P<0.05);与H/R+RLMTF+anti-miR-NC组比较,H/R+RLMTF+anti-miR-1247-3p组MDA水平、细胞凋亡率升高(P<0.05),SOD、GSH-Px活性降低(P<0.05)。结论:金樱子总黄酮通过促进miR-1247-3p表达,抑制氧化应激及细胞凋亡,从而减轻H/R诱导的心肌细胞损伤。

金樱子的抗菌活性及其有效部位研究

比较金樱子不同提取方法和不同溶媒萃取部位的体外抗菌效果,确定金樱子抗菌物质最有效的提取方式和有效部位。以金樱子果实为原料,采用甲醇冷浸法、乙醇热回流法提取,得到金樱子粗提物总浸膏,再依次用石油醚、乙酸乙酯和水对粗提物总浸膏依次萃取,得到不同溶媒的萃取部位,然后利用96微孔板稀释实验法进行体外抗菌实验。两种提取方法得到的粗提物对大多数细菌表现出一定的抗菌作用,甲醇冷浸法得到的粗提物的抑菌效果优于乙醇回流法。石油醚萃取部位和乙酸乙酯萃取部位呈现出良好的抗菌效果,水萃取部位抗菌效果较差。其中甲醇冷浸法获得的石油醚萃取部位对枯草芽孢杆菌CMCC 63501的抗菌活性最强,最低抑菌浓度达到0.1562 mg/mL。乙醇回流法所得的石油醚萃取部位对变异链球菌BNCC 336931的抑菌作用最强,其最低抑菌浓度达到0.3125 mg/mL。金樱子果具有广谱的抗菌活性,初步确定金樱子抗菌有效部位主要是甲醇冷浸提取法和乙醇热回流法所得的石油醚、乙酸乙酯萃取部位。