Mechanism of Rosae Rugosae Flos flavonoids in the treatment of hyperlipidemia and optimization of extraction process based on network pharmacology

This study aims to identify a natural plant chemical with hypolipidemic effects that can be used to treat high cholesterol without adverse reactions.Through network pharmacology screening,it was found that Rosae Rugosae Flos(RF)flavonoids had potential therapeutic effects on hyperlipidemia and its mechanism of action was discussed.TCMSP and GeneCards databases were used to obtain active ingredients and disease targets.Venn diagrams were drawn to illustrate the findings.The interaction network diagram was created using Cytoscape 3.8.0 software.The PPI protein network was constructed using String.GO and KEGG enrichment analysis was performed using Metascape.The results revealed 2 active flavonoid ingredients and 60 potential targets in RF.The key targets,including CCL2,PPARG,and PPARA,were found to play a role in multiple pathways such as the AGE-RAGE signaling pathway,lipid and atherosclerosis,and cancer pathway in diabetic complications.The solvent extraction method was optimized for efficient flavonoid extraction based on network pharmacology prediction results.This was achieved through a single factor and orthogonal test,resulting in an optimum process with a reflux time of 1.5 h,a solid-liquid ratio of 1:13 g/mL,and an ethanol concentration of 50%.

Exploring the mechanism of Crocus sativus and Rosa rugosa for the treatment of coronary heart disease based on network pharmacology and molecular docking

Coronary atherosclerotic heart disease(CHD)is the main type of cardiovascular disease.The efficacy of Uyghur drug compound Saffron formula in CHD has been clinically proven.However,the underlying mechanism remains unclear.In this study,researchers investigated the active ingredients and mechanism of action of Crocus sativus and Rosa rugosa in the treatment of CHD by network pharmacology and molecular docking techniques,collected target information with the help of TCMSP,GEO,GeneCards,and other databases,constructed protein-protein interaction(PPI)network diagrams by STRING database,performed GO and KEGG pathway enrichment analysis on common targets,and finally molecularly docked the active ingredients with core targets.C.sativus-R.rugosa have a variety of polyphenol compounds,a total of 12 active ingredients,including quercetin and kaempferol,were screened.The first three targets intersected with the core targets of CHD as AKT1,TNF,and IL-1B.Enrichment results of KEGG pathway showed that C.sativus-R.rugosa against CHD involved atherosclerosis pathways.The molecular docking results showed that quercetin and kaempferol were well bound to the core targets,and it was speculated that these components might be the main active ingredients for the treatment of CHD.The potential mechanism of action of C.sativus-R.rugosa for the treatment of coronary heart disease was initially revealed.

Conjugation of Candida rugosa lipase with hydrophobic polymer improves esterification activity of vitamin E in nonaqueous solvent

We described a novel polymer-lipase conjugate for high-efficient esterification of vitamin E using vitamin E and succinic anhydride as the substrates in nonaqueous media.In this work,the monomer,N-isopropylacrylamide(NIPAM),was grafted onto Candida rugosa lipase(CRL)to synthesize poly(NIPAM)(pNIPAM)-CRL conjugate by atom transfer radical polymerization via the initiator coupled on the surface of CRL.The result showed that the catalytic efficiencies of pNIPAM-CRL conjugates(19.5-30.3 L·s^(-1)·mmol^(-1))were at least 7 times higher than that of free CRL(2.36 L·s^(-1)·mmol^(-1))in DMSO.It was attributed to a significant increase in Kcat of the conjugates in nonaqueous media.The synthesis catalyzed by pNIPAM-CRL co njugates was influenced by the length and density of the grafted polymer,water content,solvent polarity and molar ratio of the substrates.In the optimal synthesis,the reaction time was shortened at least 7 times,and yields of vitamin E succinate by pNIPAM-g-CRL and free CRL were obtained to be 75.4%and 6.6%at 55℃after the reaction for 1.5 h.The result argued that conjugation with pNIPAM induced conformational change of the lid on CRL based on hydrophobic interaction,thus providing a higher possibility of catalysis-favorable conformation on CRL in nonaqueous media.Moreover,pNIPAM conjugation improved the thermal stability of CRL greatly,and the stability improved further with an increase of chain length of pNIPAM.At the optimal reaction conditions(55℃and 1.5 h),pNIPAM-g-CRL also exhibited good reusability in the enzymatic synthesis of vitamin E succinate and kept~70%of its catalytic activity after ten consecutive cycles.The research demonstrated that pNIPAM-g-CRL was a more competitive biocatalyst in the enzymatic synthesis of vitamin E succinate and exhibited good application potential under harsh industrial conditions.

界面活化的溶胶凝胶包埋Candida rugosa脂肪酶催化合成维生素E琥珀酸酯

采用界面活化的溶胶凝胶包埋Candida rugosa脂肪酶(CRL)催化合成了维生素E琥珀酸酯. 考察了影响溶胶凝胶包埋固定化CRL的因素, 获得的最佳固定化条件为: 丙基三甲氧基硅烷/正硅酸四乙酯摩尔比为1/1, 水与硅烷前体摩尔比为15, 酶的添加量为0.5mg/ml, PEG400的添加量为12μl/ml溶胶. 溶胶凝胶包埋的CRL在50℃, 18h后其活性仍然保持了70.58%, 是游离酶的2.6倍,且稳定性得到了明显的改善. 基于CRL的界面特性, 采用五种表面活性剂对其进行界面活化. 结果表明, 采用橄榄油活化的溶胶凝胶包埋的CRL合成维生素E琥珀酸酯的酯化活力最高, 相比原酶和未界面活化的溶胶凝胶包埋酶分别提高了6.7和1.43倍.

脂肪酶产生菌Candida rugosa产酶条件研究

We studied some factors affecting the lipase production from candida rugosa, they mainly included medium compositions and culture condition.The result showed that the optimal medium compositions for lipase production are 0.1% glucose 4.0% olive oil (carbon source),0.3% NH 4NO 3(nitrogen source),1.2%K 2HPO 4 and 0.4%MgSO 4·7H 2O.And the optimal culture condition is initial pH6.5,temperature 30℃,agitation 180r/min and time 60h. As a result, and the lipase activity could reach 19.5u/mL.Meanwhile we found that the surfactant could be helpful to the lipase production, and the optimal surfactant concentration was 0.03% GPE.The lipase activity was improved by more than 170% after we optimized the medium compositions and culture condition.While in a 5L fermentator, the lipase activity of fermentation broth could reach 33.5u/mL within 48 hours.

Diterpenoids from the Roots of Agastache rugosa

Agastol (2), a new diterpene, was isolated from the roots of Agastache rugosa together with its isomer, named isoagastol (3) Their structures were established on the basis of spectral methods The structures of agastol (2) and isoagastol (3) was elucidated as 11,14 dihydroxy 12 methoxy 19(4→3) abeo abieta 4(18),8,11,13 tetraen 7 one and 11,14 dihydroxy 12 methoxy 19(4→3) abeo abieta 3,8,11,13 tetraen 7 one Isoagastol was isolated for the first time from natural sources

‘紫枝’玫瑰(Rosa rugosa ‘Zi zhi’)开花过程花青素相关化合物及代谢途径分析

【目的】分析‘紫枝’玫瑰5个开花时期花瓣中的花青苷、类黄酮苷和类胡萝卜素的种类和含量,推定‘紫枝’玫瑰花的花青苷代谢途径,为探讨玫瑰花色的呈色机理和花色育种提供参考。【方法】以‘紫枝’玫瑰不同开花时期的花瓣为试材,用高效液相色谱(HPLC)和超高效液相色质联用分析法(UPLC-DAD-Q-TOF-MS)对其花青苷、类黄酮苷和类萝卜素进行结构推定和定量分析,结合化学反应过程推测‘紫枝’玫瑰花青苷代谢途径。【结果】在‘紫枝’玫瑰中鉴定出8种花青苷、16种类黄酮苷和β-胡萝卜素,没有检测到叶黄素;花青苷主要以芍药素、飞燕草素、矢车菊素和天竺葵素的双糖苷为主,四类花青苷都在花蕾期和初开期相对含量最高;检测到芍药苷的两种甲基化衍生物,没有发现飞燕草苷的甲基化衍生物;类黄酮苷以槲皮素和山萘酚的糖苷化、酰基化和甲基化的衍生物为主。定量分析显示,芍药苷和飞燕草苷占‘紫枝’玫瑰总花青苷含量的90%以上;芍药苷含量在‘紫枝’玫瑰开花过程中随花色变浅而降低,飞燕草苷在开花过程含量变化不大;芍药苷和飞燕草苷的比例(Pn/Dp)随花色变浅而降低。【结论】‘紫枝’玫瑰花中含有芍药素、飞燕草素、矢车菊素和天竺葵素,这四类花青素在半开期之前完成全部积累,盛开期后只降解不合成。芍药素是形成‘紫枝’玫瑰花色的主要成分。‘紫枝’玫瑰的花青苷代谢途径中,甲基酶(Rr AOMT)的催化作用有底物特异性。

胶东海岸野生玫瑰(Rosa rugosa)的濒危现状与保护策略

玫瑰是具有重要种质和生态价值的濒危植物 ,胶东海岸是玫瑰在我国的主要分布区之一 .通过野外调查和种群特征分析 ,基本查清了该区域内现存的玫瑰野生分布区 (点 )及主要分布区的生境特征、玫瑰种群的数量、空间格局 ,揭示了人为导致的生境破坏、玫瑰种子传播受阻是导致野生玫瑰种群数量持续减少、有性繁殖成功率低的主要原因 .在此基础上 ,提出应对该区域的野生玫瑰种群采取就地保护、迁地保护和利用性保护相结合的多途径保护策略 .

皱褶假丝酵母Candida rugosa产脂蛋白酯酶的发酵培养基优化

将筛选得到的产脂蛋白酯酶的皱褶假丝酵母进行了底物诱导和发酵培养基的优化,确定了其最适培养基配方:橄榄油0.6%,蛋白胨0.5%,酵母膏0.3%,麦芽浸膏0.3%,Tween-80 0.1%,K2HPO4 0.05%,Na2HPO4 0.1%,MgSO4.7H2O 0.1%。在最适培养基中培养,脂蛋白酯酶的酶活力可达121.65U/L,比未优化前提高了80倍。

A study on JA-and BTH-induced resistance of Rosa rugosa‘Plena’to powdery mildew(Sphaerotheca pannosa)

Different concentrations of jasmonic acid(JA)and benzothiadiazole(BTH) were sprayed on 2-year-old Rosa rugosa‘Plena’ seedlings. The induced resistance of JA and BTH to Sphaerotheca pannosa(Wallr.) and the changes of their related physiological indices were investigated. Results showed that JA and BTH treatments had inhibitory impacts on S. pannosa infection. The optimal concentration of JA and BTH was 0.5 mmol/L for the disease-resistance induction of the leaves, its inductive effect was up to 66.36% for BTH and 54.49% for JA. Our results confirmed that exogenous JA and BTH significantly improved R. rugose ‘Plena’ resistance to S. pannosa. When treated with JA and BTH, activities of the three defense enzymes(POD, PPO, and PAL) increased significantly.Contents of total phenolics, flavonoids, and lignin also increased significantly. It is inferred from these results that exogenous JA and BTH could improve the resistance of R.rugose ‘Plena’ to S. pannosa through enhancing activities of the defensive enzymes and accumulation of secondary metabolites in the leaves.

Candida rugosa脂肪酶催化(R,S)-2-辛醇酯交换反应的对映选择性

考察了皱落假丝酵母脂肪酶(CRL)在有机溶剂、反胶束和离子液体中催化(R,S)-2-辛醇不对称酯交换反应的对映选择性,发现反应介质和酰基供体对CRL的对映选择性有显著的影响。在甲苯中,CRL催化氯乙酸乙酯与(R,S)-2-辛醇酯交换反应的对映体选择率E颇高(E=92.2);在CTAB/丁醇/异辛烷/水反胶束体系中,以乙酸乙烯酯为酰基供体时,CRL的对映选择率很高(E》100);在离子液体1-丁基-3-甲基咪唑六氟磷酸盐中,CRL的对映选择率很低。

The Grey Analysis, Kriging and Selection Index of Flower Yield in Rugosa Rose

The analysis of grey system, kriging interpolation, and integration selection index were employed to investigate the relationships between the flower yield/plant （FY） and 15 other quantitative traits of 20 rugosa rose cultivars. The result showed that： The grey relational grade （GRG） of the number of flowers/plant （NF）, the number of branches/plant （NB）, the width of floral bud （WB）, and the weight/flower （WF） to the FY were larger （〉 0.5）; FY improved with the increase of NF and NB. Moreover, the indirect selection of either trait could not achieve improvement of FY. It is necessary to improve FY by multi-trait selection. The integration selection index （ISI） equation of FY was established with the characters NF, NB, WB, and WF： I= 0.3187x1 - 318.6x2 ＋ 670.1 x4 ＋ 6.3xa, index heritability = 0.8014, selective response of the integration breeding value = 245.8811. This will provide a theoretic basis for the genetic breeding of rugosa rose.

Candida rugosa脂肪酶交联酶晶体稳定性研究

交联酶晶体（ＣｒｏｓｓＬｉｎｋｅｄＥｎｚｙｍｅＣｒｙｓｔａｌ，ＣＬＥＣｓ）是一种新型的生物催化剂．本研究对Ｃａｎｄｉｄａｒｕｇｏｓａ脂肪酶交联酶晶体的制备以及其稳定性进行了探索，制备得的交联酶晶体不仅保持酶原有的特性，而且提高了酶在极端环境条件（高酸碱度、高温）下的稳定性。

Stability of Surfactant-coated Candida Rugosa Lipase in Isooctane

The stability of Candida rugosa lipase coated with glutamic acid didodecyl ester ribitol amide was investigated taking esterification of lauryl alcohol and lauric acid in isooctane as a model reaction. At 30C, the half-life of the activity of the coated lipase was ca 10 h, the enzyme activity became less changed after 12 h and the residual activity was 39% of the initial value. The coated lipase obeyed a first-order deactivation model with a deactivation energy of 29.9J.mol-1.

Combination Features,Paleobiogeographic Affinity and Mass Extinction of the Latest Ordovician(Hirnantian) Rugosan Fauna from Northern Guizhou,China

The rugosan fauna from the Guanyinqiao Bed （latest Ordovician, Hirnantian） of northern Guizhou, China is known to belong to the cold or cool-water type corals. The components of the fauna are solitary corals only, and corallite septa are generally strongly dilated, especially the streptelasmatid corals are dominant comprising 98% of the whole fauna. The Guanyinqiao Bed is rich in rugosans of 18 genera, which are streptelasnmtid Streptelasma （=Helicelasma）, Brachyelasma, Amplexobrachyelasma, Salvadorea, Grewingkia, Borelasma, CrassUasma, Leolasma, KenophyUum, UUernelasma, Paramplexoides, Siphonolasma, Pycnactoides, Dalmanophyllum, Bodophyllum, Axiphoria, Lambeophyllum and cystiphyllid Sinkiangolasma. Although this fauna was fairly abundant in a confined area （northern-northeastern Guizhou, southern Sichuan） during the Hirnantian age, the rugosan mass extinction （generic extinction rate 81%） happened at the end of the Hirnantian Stage. It is conduded that the mass extinction is related to the ending of maximum glaciation and ice cap melting in Gondwana in the southern hemisphere in the latest Hirnantian, resulting in rapid global sea-level rise in the earliest Silurian. In the Upper Yangtze Basin, the sea bottom environments were replaced by anoxic and warmer water during that time, so that the cool-water type rugosan became extinct. The present paper attempts to revise some already described rugose coral genera and species （He, 1978, 1985） and to supplement a few new forms from the Guanyinqiao Bed. Fourteen species of 12 genera are re-described and illustrated, of which one species- Grewingkia latifossulata is new. As a whole, the rugosan fauna of the Guanyinqiao Bed may be correlated with those contemporaneous of North Europe, Estonia and North America, indicating a dose biogeographic affinity to North Europe.

昆明二村下奥陶统红石崖组顶部Cruziana rugosa遗迹化石群落的沉积环境和生态特征

Cruziana及其相关的遗迹化石是由底栖三叶虫在沉积物表面停歇、爬行所留下的遗迹,一般认为产于潮下带至浅海陆棚环境,具有古水深指示意义。然而,本文通过对昆明民富二村下奥陶统红石崖组顶部含Cruziana rugosa遗迹化石群落层位的沉积环境分析却发现,Cruziana rugosa遗迹化石群落的沉积环境为潮上带—潮间带,说明三叶虫可以在短暂暴露环境中活动。三叶虫的这种适应性活动可能为节肢动物从海洋走向陆地做好了进化准备。

Genetic diversity of the endangered species Rosa rugosa Thunb.in China and implications for conservation strategies

Rosa rugosa Thunb. is one of the dominant and important shrub species in estuary dunes and shingle beaches of northern China. However, its area of distribution, the number of populations, and the size of each population have decreased rapidly in the past two decades because of habitat degradation and loss. Random amplified polymorphic DNA markers were used to determine the genetic diversity of four remaining large natural populations of R. rugosa and to discuss an effective conservation strategy for this endangered species in China. High genetic variations were detected in R. rugosa populations in China. The mean percentage of polymorphic loci （P%） within four local populations was 57.99%, with the P% of the total population being 75.30%. Mean Shannon＇s information index （H0） was 0.2826, whereas total Ho was 0.3513. The genetic differentiation among populations was 0.1878, which indicates that most genetic diversity occurs within populations. Population Tumenjiang （TMJ） showed the highest genetic diversity （P% = 66.27%; H0 = 0.3117） and contained two exclusive bands. Population Changshandao （CSD） showed higher genetic diversity （P% =59.04%; H0 = 0.3065）. Populations TMJ and CSD contained 95.33% and 99.33%, respectively, of loci with moderate to high frequency （P〉0.05） of the total population. These results indicate that populations TMJ and CSD should be given priority for in situ conservation and regarded as seed or propagule sources for ex situ conservation. The results of the present study also suggest that R. rugosa in China has become endangered as a result of human actions rather than genetic depression of populations; thus, human interference should be absolutely forbidden in R. rugosa habitats.

Effect of jasmonate treatments on leaves of Rosa rugosa‘Plena’and detoxification enzymes and feeding of adult Monolepta hieroglyphica

To study the effect of jasmonates(JAs)on the resistance of economic forest plants to insects,R osa rugosa‘Plena'leaves were treated with 1 mmol/L jasmonic acid(JA),methyl jasmonate(MeJA)and Z-jasmone,then the content of tannin and total phenol in leaves and the feeding area of Monolepta hieroglyphica adults on leaves were measured.Changes in the activities of detoxification enzymes in adult M.hieroglyphica that had fed on leaves treated with JAs were also studied.Tannin and total phenol levels in leaves increased significantly after treatment with JAs,and tannin level was 1.36–1.55-fold higher than in the control after treatment with 1 mmol/L MeJA.The total content of phenol in leaves treated with 1.0 mmol/L Z-jasmone increased by 1.33–2.20-fold compared with those of the control.The activities of detoxification enzymes in adults were inhibited to differing degrees:activity of alkaline phosphatase(AKP)first increased,then decreased;the activities of acid phosphatase(ACP),glutathione S-transferases(GSTs)and carboxylesterase(CarE)following treatment with 1 mmol/L MeJA were significantly reduced and were 22%–31%,11%–26%,and 11%–31%lower than those of the control,respectively.Moreover,the feeding area of adult M.hieroglyphica on the leaves treated with JAs was significantly reduced(P<0.05).The feeding area of economic forest R.rugosa‘Plena'leaves treated with 1 mmol/L MeJA decreased by 17%–43%compared with that of the control.Moreover,the decrease in the adult M.hieroglyphica feeding area was highly positively correlated with the content of tannin and positively correlated with the contents of total phenol of economic forest R.rugosa‘Plena'leaves.The reduced feeding area of adult M.hieroglyphica was highly negatively correlated with the activities of AKP and ACP and negatively correlated with those of the GSTs.In conclusion,the use of 1 mmol/L MeJA can noticeably decrease the deleterious effects of adult M.hieroglyphica.

Cloning and Expression Analysis of <i>RrGT2</i>Gene Related to Anthocyanin Biosynthesis in <i>Rosa rugosa</i>

At present, the research about flower color of Rosa rugosa is a very inno-vative and practical study. Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1422bp, encoding 473 amino acids, designated as RrGT2, were isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT2 gene is C2334H3628N602O711S18, the relative molecular mass is 52,075.17 Da, and the theoretical isoelectric point is pI = 4.76. The result of the RrGT2 protein 3D model construction showed that it had the highest homology with the UDP-glycosyltransferase 74F2 protein model in the database (39.53%). Sequence alignments with the NCBI database showed that the RrGT2 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT2 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT2 enzyme activity. RrGT2 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of antho-cyanins. Therefore, we speculated that glycosylation of RrGT2 plays a crucial role in anthocyanin biosynthesis in R. rugosa.

Component Analysis of Anthocyanins in Petals at Different Flowering Stages of Three <i>Rosa rugosa</i>Hybrid Cultivars

The species and contents of anthocyanins in plant petals can make plants appear pink, red, violet and blue, etc., and play a major role in the coloration of plants. In this study, the species and contents of anthocyanins in the petals of three R. rugosa hybrid cultivars, R. rugosa “Zizhi”, R. rugosa “Fenzizhi” and R. rugosa “Baizizhi”, were analyzed, and the direct cause of the differences in flower color of three R. rugosa hybrid cultivars was inferred. This paper provides a reference for the coloration mechanism and flower color breeding of R. rugosa. The specific methods are as follows: the petals of five flowering stages of three R. rugosa hybrid cultivars were used as materials, and the types and contents of anthocyanins contained in them were qualitatively and quantitatively analyzed by high performance liquid chromatography (HPLC). The same six kinds of anthocyanins were identified in R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, mainly based on the diglycoside of paeoniflorin and cyanidin. The relative contents of the two anthocyanins were higher at budding stage and initial opening stage. In the different flowering stages of R. rugosa “Zizhi”, the content of Pn3G5G was up to 4280.84 ± 20.82 μg&middot;g-1, and the content of Cy3G5G was up to 789.41 ± 1.21 μg&middot;g-1. In R. rugosa “Fenzizhi”, the highest content of Pn3G5G reached 1293.50 ± 17.64 μg&middot;g-1, and the content of Cy3G5G was up to 358.86 ± 3.94 μg&middot;g-1. It could be speculated that the difference in the contents of Pn3G5G and Cy3G5G was the main reason for the difference in coloration between the petals of R. rugosa “Zizhi” and R. rugosa “Fenzizhi”. A total of five species of anthocyanins were identified in R. rugosa “Baizizhi” and their contents were relatively low. Compared with R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, the presence of Cy3G was not detected. Therefore, we speculated that the two reasons above might be responsible for the visual white flowers of R. rugosa “Baizizhi”.