High-resolution monophenolase/diphenolase/radical scavenging profiling for rapid screening of natural whitening candidates from Rosa rugosa cv. 'Plena'.

Antioxidants and tyrosinase inhibitory components were successfully screened and separated from Rosa rugosa cv. 'Plena' by high-performance liquid chromatography microfractionation bioactive screening combined with several separation and purification methods. Ethyl acetate extract of Rosa rugosa cv. 'Plena' showed high antioxidant activity and tyrosinase inhibitory activity. High-speed countercurrent chromatography, silica gel column chromatography, and semi-preparative high-performance liquid chromatography were used for the preparative separation of four bioactive components from ethyl acetate extract. Two tyrosinase-inhibiting active substances, flavogallonic acid, and N1 -N5 -N10 -tri-4-p-coumaroylspermidine, were isolated from Rosa rugosa cv. 'Plena', and they showed great monophenolase inhibition activity (half-maximal inhibitory concentration: 664.60 and 23.77 µg/ml, respectively) and excellent diphenolase inhibition activity (half-maximal inhibitory concentration: 23 614.61 and 16.80 µg/ml, respectively). Meanwhile, gallic acid, flavogallonic acid, and ellagic acid were shown to have excellent 1,1-diphenyl-2-picryl-hydrazyl antioxidant activity (half maximal inhibitory concentration: 6.66, 20.17, and 13.45 µg/ml), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) antioxidant activity (half maximal inhibitory concentration: 3.53, 3.83, and 2.78 µg/ml). Molecular docking revealed that flavogallonic acid and N1 -N5 -N10 -tri-4-p-coumaroylspermidine had a strong binding affinity (-9.3 and -10 kcal/mol, respectively) to tyrosinase through hydrogen bonding and hydrophobic interactions.

BOX38, a DNA Marker for Selection of Essential Oil Yield of Rosa × rugosa.

Rosa rugosa L. was a famous aromatic plant whose cultivars (Rosa × rugosa) have been widely used in the perfume industry in Asia. The perfume market looks for rose cultivars bearing higher essential oil, while the oil yields of most R. × rugosa have not been evaluated due to limiting conditions, such as insufficient cultivation areas. Here, we tested the yield and the aroma components of essential oil of 19 R. × rugosa. The results indicated that the yields of nerol, citronellol, and geraniol could represent an alternative index of the total yield of essential oil. Sequence syntenic analysis indicated that the Rosa genus specific cis-element Box38 was highly polymorphic. The Box38 region isolation of Rosa × rugosa by flanked primers proved that Box38 repeat number was significantly positively correlated with the essential oil yield of the corresponding cultivar. In the breeding of Rosa × rugosa, six-Box38-repeat could be a robust threshold for selection of high-essential-oil roses. Together, we found that Box38 was a DNA marker for essential oil yield and that it would be helpful in the early selection and breeding of essential oil roses.

GT Transcription Factors of Rosa rugosa Thunb. Involved in Salt Stress Response.

Rosa rugosa was a famous aromatic plant while poor salt tolerance of commercial cultivars has hindered its culture in saline-alkali soil. In many plants, the roles of GT (or trihelix) genes in salt stresses responses have been emerging. In the wild R. rugosa, a total of 37 GTs (RrGTs) were grouped into GT-1, GT-2, GTγ, SH4, and SIP1 lineages. SIP1 lineage expanded by transposition. The motifs involved in the binding of GT cis-elements were conserved. Four RrGTs (RrGT11/14/16/18) significantly differentially expressed in roots or leaves under salt stress. The responsive patterns within 8 h NaCl treatment indicated that RrGTγ-4 (RrGT18) and RrGT-1 (RrGT16) were significantly induced by salt in roots of R. rugosa. Subcellular localizations of RrSIP1 (RrGT11) and RrGTγ-4 were on chloroplasts while RrGT-1 and RrSIP2 (RrGT14) located on cell nucleus. Regulation of ion transport could be the most important role of RrSIPs and RrGTγ-4. And RrGT-1 could be a halophytic gene with higher transcription abundance than glycophytic GT-1. These results provide key clue for further investigations of roles of RrGTs in salt stress response and would be helpful in the understanding the salt tolerance regulation mechanism of R. rugosa.

Systematic Identification and Analysis of OSC Gene Family of Rosa rugosa Thunb.

The oxidosqualene cyclase family of Rosa rugosa (RrOSC) provides a starting point for the triterpenoid pathway, which contributes to the medicinal value of the extraction of tissues of Rosa rugosa. However, the structure and function of key RrOSCs of active triterpenoids remain ambiguous. In this study, a total of 18 RrOSC members with conservative gene structures and motifs were identified based on the genome of Rosa rugosa. The RrOSCs were located on three chromosomes including two gene clusters that derived from gene replication. The phylogenetic relationship divided RrOSCs into six groups, and the RrOSCs of GI and GIV that were represented by lupeol or α-amyrin were identified as likely to include candidate genes for producing active triterpenoids. Considering the high expression or specific-tissue expression of the candidates, RrOSC1, RrOSC10, RrOSC12, and RrOSC18 were considered the key genes. RrOSC12 was identified in vitro as lupeol synthase. The results provided fundamental information and candidate genes for further illustration of the triterpenoid pathway involved in the pharmacological activities of Rosa rugosa.

Rosa rugosa polysaccharide induces autophagy-mediated apoptosis in human cervical cancer cells via the PI3K/AKT/mTOR pathway.

The aim of this study was to investigate the effect of a polysaccharide from Rosa rugosa Thunb. on human cervical cancer cells (HCCCs) and the underlying mechanism. Here, a novel Rosa rugosa polysaccharide, named as RRP, was purified from Rosa rugosa petals. RRP consisted of glucose, galacturonic acid, mannose, rhamnose, galactose, arabinose, xylose, and glucuronic acid (molar ratio: 7.78:7.59:4.23:3.22:3.15:1.65:1.00), with Mw of 327.92 kDa. RRP remarkably inhibited cell proliferation, migration, and cell cycle arrest in HeLa and SiHa cells. Furthermore, RRP induced apoptosis by activating the caspase family of proteins and mediating the reactive oxygen species (ROS)-mediated mitochondrial pathway. In addition, RRP was found to dose-dependently induce autophagy, which occurred prior to apoptosis. RRP also primarily induced autophagy-mediated apoptosis in HCCCs via the PI3K/AKT/mTOR pathway. Thus, RRP might serve as a legitimate therapeutic drug candidates against human cervical cancer.

Analysis of microRNAs, phased small interfering RNAs and their potential targets in Rosarugosa Thunb.

BACKGROUND: MicroRNAs (miRNAs) are small non-coding RNAs that play important roles by regulating other genes. Rosa rugosa Thunb. is an important ornamental and edible plant, yet there are only a few studies on the miRNAs and their functions in R. rugosa. RESULTS: We sequenced 10 samll RNA profiles from the roots, petals, pollens, stamens, and leaves and 4 RNA-seq profiles in leaves and petals to analysis miRNA, phasiRNAs and mRNAs in R. rugosa. In addition, we acquired a degradome sequencing profile from leaf of R. rugosa to identify miRNA and phasiRNA targets using the SeqTar algorithm. We have identified 321 conserved miRNA homologs including primary transcripts for 25 conserved miRNAs, and 22 novel miRNAs. We identified 592 putative targets of the conserved miRNAs or tasiRNAs that showed significant accumulations of degradome reads. We found differential expression patterns of conserved miRNAs in five different tissues of R. rugosa. We identified three hundred and thirty nine 21 nucleotide (nt) PHAS loci, and forty nine 24 nt PHAS loci, respectively. Our results suggest that miR482 triggers generations of phasiRNAs by targeting nucleotide-binding, leucine-rich repeat (NB-LRR) disease resistance genes in R. rugosa. Our results also suggest that the deregulated genes in leaves and petals are significantly enriched in GO terms and KEGG pathways related to metabolic processes and photosynthesis. CONCLUSIONS: These results significantly enhanced our knowledge of the miRNAs and phasiRNAs, as well as their potential functions, in R. rugosa.

Ultrasound-assisted extraction and antioxidant activity of phenolic and flavonoid compounds and ascorbic acid from rugosa rose (Rosa rugosa Thunb.) fruit.

The present study aimed to extract total phenolic compounds (TPC), total flavonoid compounds (TFC), and ascorbic acid (AA) from the fruit of rugosa rose (Rosa rugosa Thunb.) by ultrasound-assisted extraction (UAE), and to evaluate their antioxidant activities. UAE significantly increased the extract yield compared with that obtained using the conventional method. TPC, TFC, and AA were extracted, depending on the extraction conditions (temperature, time, and ethanol concentration), in the range of 50.73-96.69, 15.93-31.88, and 3.06-6.08 mg/g, respectively. TPC and TFC were effectively extracted at a relatively high temperature (50 °C) than AA was (30 °C). The solvent condition used to extract TPC, TFC, and AA was 50% ethanol. The UAE condition for the highest antioxidant activity was obtained 30 °C, 30 min, and 50% ethanol, which were the same condition for the highest AA extraction. Among the extracts, AA showed a strong correlation with antioxidant activity at p-value of 0.001.

Polyphenol-enriched extract of Rosa rugosa Thunb regulates lipid metabolism in diabetic rats by activation of AMPK pathway.

This study was designed to investigate the mechanism of polyphenol-enriched extract of Rosa rugosa Thunb (RPE) in the control of dyslipidemia in diabetic rats. RPE was tested at three dosages (37.5 mg/kg, 75 mg/kg and 150 mg/kg) in the rat dyslipidemia model established with high fat diet feeding in combination with STZ injection (30 mg/kg). The RPE effect was evaluated after 4 weeks of treatment. In the RPE-treated rats, hepatic total cholesterol (TC) and triglyceride (TG) were significantly reduced, lipoprotein lipase (LPL) and liver lipase (HL) were significantly increased. The levels of alanine transaminase (ALT) and aspartate transaminase (AST) were decreased in the serum. Those effects of RPE were observed primarily at the mediate and high dosages. Expression of FGF21 was increased in the liver tissue and hepatic cell line 1c1c7 by RPE. The signals of p-AMPK, p-ACC, ACC, p-SIRT, and PGC-1α were significantly induced in the liver by RPE. The results suggest that RPE may improve hepatic steatosis and liver function by induction of AMPK signaling activity in the control of dyslipidemia.

Effects of Uygur Medicine Branchlets roses extracts on blood glucose and glucose tolerance in diabetes mice induced by alloxan / 药物评价研究

Objective To observe the effect of extract from Branchlets roses on blood glucose and glucose tolerance in diabetes mice induced by alloxan.Methods Diabetes animal model was established by alloxan.Dividing the model mice into eight groups:model group,water extract high,middle,and low dose (3.70,1.85,and 0.93 g/kg) group,and ethanol extract high,middle,and low dose (2.75,1.37,and 0.70 g/kg) group,and metformin (positive drug,200 mg/kg) group,and normal mice were taken as control group.Drug was ig administered to mice 3 d after molding once daily.Blood glucose test paper was used to determine fasting blood glucose 0,10,20,and 28 d after modeling,and the glucose tolerance test was performed 30 d after modeling.Results The extract of Branchlets roses from all the groups could decrease the blood glucose and improve the glucose tolerance,and showed a certain dose-effect relationship.In all the extracts,the alcohol extract had the best effect,but the effect was not as good as the positive control drug metformin hydrochloride group.Conclusion The extract of Branchlets roses can reduce the blood sugar content of diabetic mice,and improve the glucose tolerance.

Anti-Inflammatory Effect of Rosa rugosa Flower Extract in Lipopolysaccharide-Stimulated RAW264.7 Macrophages.

Rosa rugosa Thunb, a deciduous shrub of the genus Rosa, has been widely used to treat stomach aches, diarrhoea, pain, and chronic inflammatorydisease in eastern Asia. In recent years, our research team has extensively studied the Rosa rugosa flowerextract, and specificallyundertook pharmacological experiments which have optimized the extraction process. Our methods have yielded a standard extract enriched in phenolic compounds, named PRE. Herein, we expand our efforts and evaluated the antiinflammatoryactivity of PRE on lipopolysaccharide (LPS)-induced inflammationin RAW 264.7 macrophages. PRE significantlyinhibited production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and interleukin 1ß (IL-1ß), as well as expression of their synthesizing enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase2 (COX-2). Furthermore, PRE inhibited activity of mitogen-activated protein kinases (MAPK) as well as nuclear factor-kappa B (NF-κB) signaling pathway. Our findingsare the firstto explain the anti-inflammatorymechanism by PRE in LPS-stimulated macrophages. Given these results, we propose that PRE has therapeutic potential in the prevention of inflammatory disorders.

ISSR analysis and identification of medicinal plants Rosa chinensis, Rosa rugosa, and their relative species / 中草药

Objective: To study the genetic diversity and genetic relationship of medicinal plants Rosa chinensis, R. rugosa, and their relative species by ISSR molecular marker technique, and provide the reference for Rosa L. germplasm identification and breeding. Methods: Fifteen species (including 33 samples) of medicinal herbs R. chinensis, R. rugosa, and their relative species were studied by ISSR-PCR markers. Nei's genetic diversity index (H) and other parameters of genetic information were calculated by POPGEN 32, and a cluster dendrogram of different samples was established based on the unweighted pair-group method with arithmetic mean (UPGMA) by NTSYS-pc software. Results: Six ISSR primers generated 110 loci of which 109 loci were polymorphic. The average percentage of polymorphie bands (PPB) was 99.09%. Nei's genetic diversity index (H) and Shannon's information index (I) were 0.370 5 and 0.546 4, and the coefficient of genetic differentiation (Gst) and gene flow (Nm) were 0.886 8 and 0.063 8 between the species levels. The genetic distance (D) varied from 0.169 1 to 0.730 2. In the cluster dendrogram, 15 species were clustered into six groups at the level of Genetic similarity coefficient (GS) 0.60. Conclusion: The results of ISSR analysis reveal that medicinal plants R. chinensis, R. rugosa and their relative species had the plentiful genetic diversity and the genetic relationships were consistent with morphological characters of taxonomy. ISSR method is efficient for identification of R. chinensis, R. rugosa, and their relative species, which could provide a scientific basis for the resource collection and identification of the species in Rosa L.

Anti-Inflammatory Effect of Rosa rugosa Flower Extract in Lipopolysaccharide-Stimulated RAW264.7 Macrophages

Rosa rugosa Thunb, a deciduous shrub of the genus Rosa, has been widely used to treat stomach aches, diarrhoea, pain, and chronic inflammatory disease in eastern Asia. In recent years, our research team has extensively studied the Rosa rugosa flower extract, and specifically undertook pharmacological experiments which have optimized the extraction process. Our methods have yielded a standard extract enriched in phenolic compounds, named PRE. Herein, we expand our efforts and evaluated the anti-inflammatory activity of PRE on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophages. PRE significantly inhibited production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-a, interleukin (IL)-6, and interleukin 1β (IL-1β), as well as expression of their synthesizing enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase2 (COX-2). Furthermore, PRE inhibited activity of mitogen-activated protein kinases (MAPK) as well as nuclear factor-kappa B (NF-κB) signaling pathway. Our findings are the first to explain the anti-inflammatory mechanism by PRE in LPS-stimulated macrophages. Given these results, we propose that PRE has therapeutic potential in the prevention of inflammatory disorders.

Anti-hyperplasia effects of Rosa rugosa polyphenols in rats with hyperplasia of mammary gland.

Rosa rugosa (Thunb.) is used in Chinese traditional medicine with the functions of promoting blood circulation, relieving the depressed liver and attenuating breast disorders. This study was to investigate the anti-hyperplasia effects of the polyphenols-rich fraction from R. rugosa (FRR) in rat. Rat model of hyperplasia of mammary gland (HMG) was induced by intramuscularly injected with estrogen (0.5mg/kg/d) for 25 days, and followed with progestogen (5mg/kg/d) for another 5 days. Meanwhile, FRR was orally given for 30 days. Then, the levels of estradiol and oxidative stress were assessed. The mammary expressions of AKT and JNK were evaluated by Western blot analysis. The expressions of NFκB-p65, COX-2 and VEGF were measured by immunohistochemical analysis. The whole results indicated that FRR could exert anti-hyperplasia effects in rat via modulating the mammary expression of JNK and AKT, as well as alleviating the NFκB related oxidative stress and inflammatory responses.

Isoflavones from flower buds of Rosa rugosa and their cytotoxicity / 中草药

Objective: To study the chemical constituents from the flower buds of Rosa rugosa. Methods: The chemical constituents from the flower buds of R. rugosa were isolated by silica gel, MCI-gel resin, Sephadex LH-20 column chromatography and high performance liquid chromatography (HPLC) methods. Their structures were elucidated by spectroscopic methods and physicochemical properties. Results: Three isoflavones, 6,8-dihydroxy-4',7-dimethoxyisoflavone (1), prunetin (2), and pratensein (3) were isolated from the flower buds of R. rugosa. Conclusion: Compound 1 is a new compound named rosa isoflavone. Compounds 2 and 3 are isolated from R. rugosa for the first time. Compound 1 displays the stronger cytotoxicity against A549 and PC3 cells with IC50 values of 2.6 and 3.2 μmol/L, respectively.

Flavonoids from flower buds of Rosa rugosa / 中草药

Objective: To identificate the bioactive natural products, and study the chemical constituents in the flower buds of Rosa rugosa. Methods: The chemical constituents in the flower buds of R. rugosa were isolated by silica gel, MCI-gel resin, Sephadex LH-20 column chromatography, and high performance liquid chromatography (HPLC) methods. Their structures were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques. Results: Nine flavonoids were isolated from the flower buds of R. rugosa, and identified as 8-acetyl-4',7-dimethoxy-6-methyl-flavone (1), kaempferol (2), kaempferol-3-O-β-D-glucopyranoside (3), kaempferol-3-O-β-D-rutinside (4), quercetin (5), quercetin-3-O-β-D-glucopyranoside (6), rutin (7), luteolin (8), and luteolin-7-O-β-D-glucopyranoside (9). Conclusion: Compound 1 is a new compound and it displays the cytotoxicity against NB4, SH-SY5Y, PC3, A549, and MCF7 cells with IC50 values of 6.8, 5.6, 2.2, 1.8, and 7.4 μmol/L, respectively. Compound 9 is isolated from R. rugosa for the first time.

A LC/QTOF-MS/MS application to investigate chemical compositions in a fraction with protein tyrosine phosphatase 1B inhibitory activity from Rosa rugosa flowers.

INTRODUCTION: Rosa rugosa flowers used as herbal medicine possess many activities. A fraction extracted by ethyl acetate exhibited strong inhibitive activity against protein tyrosine phosphatase 1B (PTP1B) in vitro. OBJECTIVE: Establish an efficient method of LC coupled to quadrupole time-of-flight (QTOF) with tandem MS/MS to investigate the compositions in the active fraction. METHODS: Chemical compositions were separated and investigated by LC/QTOF-MS/MS in negative electrospray ionisation (ESI) mode at different collision energy (CE) values. The maximal structural information was obtained for the identification of components. RESULTS: A total of 75 compounds including tannins, their related compounds and flavonoids were identified or partially characterised according to accurate mass and the characteristic fragments at low and high CE. Meanwhile, the fragmentation pathways of gallotannins and ellagitannins (hexahydroxydiphenoyl group and lactonised valoneoyl group) were studied and proposed and were used to trace tannins in crude extracts. CONCLUSION: The results suggest that this fraction is a source of PTP1B inhibitory activity with a potential for treating diabetes.

Investigation of antiradical activity of plant material by thin-layer chromatography with image processing.

A novel, easy, and cheap technique for preliminary quantitative evaluation of antiradical activity, based on HPTLC, has been proposed. This method combines chromatographic separation of polar compounds, present in plant extracts, with data analysis by means of image processing software. Bleaching of the purple DPPH colour, caused by substances with antiradical activity, was observed and recorded using a photo camera. ImageJ, a free and open source image processing program was used for quantitative measurements. For evaluation of assay efficiency, the antiradical activity of rose flower extracts (from Rosa rugosa Thunb.) was expressed as Standard Activity Coefficients (SACs), which are relative measures of the activity to the four well known antioxidants; i.e., quercetin, gallic acid, protocatechuic acid, and Trolox. The method uses small amounts of free radical and is easily applicable - only a digital camera with freely available open source software is required.

ANALYSIS OF NUTRITION COMPONENTS IN THE FRUITS OF WILD ROSA RUGOSA THUNB / 营养学报

This paper deals with the nutrition components analysis on the fruits of wild Rosa rugosa Thunb. The results showed that total sugar was 8.21%, total acid 1.08%, protein 0.89%, pectin 1.43%, tannin 1.63%, vitamin C 1857-2027 mg/100g. The contents of 16 kinds of inorganic components and 16 kinds of amino acids were assayed too.