Rosmarinic acid improves tracheal smooth muscle responsiveness and lung pathological changes in ovalbumin-sensitized rats

Objective:To evaluate the effect of rosmarinic acid on tracheal smooth muscle responsiveness and lung pathological changes in ovalbumin-sensitized rats.Methods:Rats were randomly divided into six groups:the control group,the asthmatic group,and the asthmatic groups treated with dexamethasone(1 mg/kg;oral gavage)or three doses of rosmarinic acid(0.5,1,and 2 mg/kg;oral gavage).For induction of asthma,rats received intraperitoneal injections and inhalation of ovalbumin.After 21 days,bronchoalveolar lavage fluid and lung samples were collected for histopathological analyses.Moreover,total and differential white blood cell counts were determined.Results:The rosmarinic acid-treated group had significantly lower tracheal smooth muscle responses to methacholine than the asthmatic group.In addition,rosmarinic acid reduced white blood cell count and the percentages of eosinophils,monocytes,and neutrophils while increasing the percentage of lymphocytes.Ovalbumin-induced lung pathological changes were significantly improved by treatment with rosmarinic acid.Conclusions:Rosmarinic acid improves tracheal smooth muscle responsiveness and lung pathological changes in ovalbumin-sensitized rats.

Preparation of Rosmarinic Acid Injection from Folium perillae

[Objective] To extract the effective component rosmarinic acid from Folium perillae,study preparation technology and quality control of rosmarinic acid injection and provide test basis for hemorheological drug development and clinic application.[Method]The coarse powder of Folium perillae(40 μm)was lixiviated with hot water and acidized to make aqueous extracts.Rosmarinic acid from the aqueous mixture was extracted with Ethyl Acetate.And Et Ac was evaporated to obtain primary product.The injection was purified by pH adjustment.Concentration of rosmarinic acid in the injection was detected by high performance liquid chromatography(HPLC).Contents of tannin,resin and oxalate were eliminated according to the Chinese Pharmacopeia(2005 edition).Stability,irritation,haemolyticus,LD50 and thermogenic substance of this injection were also detected.[Result]The content of rosmarinic acid in Folium perillae injection is 2.81 mg/ml,and the content of LD50 was 406.82 mg/kg.This injection was consistent with the stipulation of Chinese Pharmacopeia(2005 edition).[Conclusion]The preparation technology of this experiment was reasonable.The stable qualities of the prepared injection meet the injection requirement.

Rosmarinic acid elicits neuroprotection in ischemic stroke via Nrf2 and heme oxygenase 1 signaling

Rosmarinic acid（RA） can elicit a neuroprotective effect against ischemic stroke, but the precise molecular mechanism remains poorly understood. In this study, an experimental ischemic stroke model was established in CD-1 mice（Beijing Vital River Laboratory Animal Technology, Beijing, China） by occluding the right middle cerebral artery for 1 hour and allowing reperfusion for 24 hours. After intraperitoneally injecting model mice with 10, 20, or 40 mg/kg RA, functional neurological deficits were evaluated using modified Longa scores. Subsequently, cerebral infarct volume was measured using TTC staining and ischemic brain tissue was examined for cell apoptosis with TUNEL staining. Superoxide dismutase activity and malondialdehyde levels were measured by spectrophometry. Expression of heme oxygenase-1（HO-1）, nuclear factor erythroid 2-related factor 2（Nrf2）, Bcl-2, Bax, Akt, and phospho-Ser473 Akt proteins in ischemic brain tissue was detected by western blot, while mRNA levels of Nrf2, HO-1, Bcl-2, and Bax were analyzed using real time quantitative PCR. In addition, HO-1 enzyme activity was measured spectrophotometrically. RA（20 and 40 mg/kg） greatly improved neurological function, reduced infarct volume, decreased cell apoptosis, upregulated Bcl-2 protein and mRNA expression, downregulated Bax protein and mRNA expression, increased HO-1 and Nrf2 protein and mRNA expression, increased superoxide dismutase activity, and decreased malondialdehyde levels in ischemic brain tissue of model mice. However, intraperitoneal injection of a HO-1 inhibitor（10 mg/kg zinc protoporphyrin IX） reversed the neuroprotective effects of RA on HO-1 enzyme activity and Bcl-2 and Bax protein expression. The PI3 K/Akt signaling pathway inhibitor LY294002（10 mM） inhibited Akt phosphorylation, as well as Nrf2 and HO-1 expression. Our findings suggest that RA has anti-oxidative and anti-apoptotic properties that protect against ischemic stroke by a mechanism involving upregulation of Nrf2 and HO-1 expression via the PI3 K/Akt signaling pathway.

Rosmarinic acid ameliorates hypoxia/ischemia induced cognitive deficits and promotes remyelination

Rosmarinic acid,a common ester extracted from Rosemary,Perilla frutescens,and Salvia miltiorrhiza Bunge,has been shown to have protective effects against various diseases.This is an investigation into whether rosmarinic acid can also affect the changes of white matter fibers and cognitive deficits caused by hypoxic injury.The right common carotid artery of 3-day-old rats was ligated for 2 hours.The rats were then prewarmed in a plastic container with holes in the lid,which was placed in 37°C water bath for 30 minutes.Afterwards,the rats were exposed to an atmosphere with 8% O2 and 92% N2 for 30 minutes to establish the perinatal hypoxia/ischemia injury models.The rat models were intraperitoneally injected with rosmarinic acid 20 mg/kg for 5 consecutive days.At 22 days after birth,rosmarinic acid was found to improve motor,anxiety,learning and spatial memory impairments induced by hypoxia/ischemia injury.Furthermore,rosmarinic acid promoted the proliferation of oligodendrocyte progenitor cells in the subventricular zone.After hypoxia/ischemia injury,rosmarinic acid reversed to some extent the downregulation of myelin basic protein and the loss of myelin sheath in the corpus callosum of white matter structure.Rosmarinic acid partially slowed down the expression of oligodendrocyte marker Olig2 and myelin basic protein and the increase of oligodendrocyte apoptosis marker inhibitors of DNA binding 2.These data indicate that rosmarinic acid ameliorated the cognitive dysfunction after perinatal hypoxia/ischemia injury by improving remyelination in corpus callosum.This study was approved by the Animal Experimental Ethics Committee of Xuzhou Medical University,China (approval No.20161636721) on September 16,2017.

Melissa officinalis and rosmarinic acid in management of memory functions and Alzheimer disease

Alzheimer disease(AD) as worldwide progressive neurodegenerative disease is prevalent disease among elderly population. Due to limitation in chemical drugs along with their adverse effects of these treatments, research for finding more effective natural drugs, is one of interesting subjects among the scientists. Melissa officinalis(M. officinalis) has a long history of use in treatment of nervous system diseases. This review article evaluates the potency of M. officinalis in treatment of AD by review of experimental and clinical documents on the efficacy, safety and its mechanisms of action in management of AD. The information is extracted from electronic resources(PubMed, Wiley, Springer and Science Direct), English and Persian scientific books. In spite of different scientific and non-scientific reports on the use of M. officinalis and its main component of rosmarinic acid in neurodegenerative diseases, there is only one clinical trial on the efficacy of M. officinalis ethanol extract in management of AD. Different mechanisms of action for M. officinalis, including inhibitory effects against amyloid beta, reactive oxygen species, and acetylcholine esterase, are involved. Larger clinical trials are recommended to confirm the efficacy and safety of M. officinalis extracts in treatment of AD patients.

Rosmarinic acid attenuates hepatic fibrogenesis via suppression of hepatic stellate cell activation/proliferation and induction of apoptosis

Objective:To investigate the antifibrotic role of rosmarinic acid(RA),a natural polyphenolic compound,on HSCs activation/proliferation and apoptosis in vitro and in vivo. Methods:The impact of RA on stellate cell line(HSC-T6) proliferation,activation and apoptosis was assessed along with its safety on primary hepatocytes. In vivo,rats were divided into:(i) normal;(ii) thioacetamide(TAA)-intoxicated rats for 12 weeks;(iii) TAA+silymarin or(iv) TAA+RA. At the end of experiment,liver functions,oxidative stress,inflammatory and profibrogenic markers,tissue inhibitor metalloproteinases type-1(TIMP-1) and hydroxyproline(HP) levels were evaluated. Additionally,liver histopathology and immunohistochemical examinations of alpha-smooth muscle actin(α-SMA),caspase-3 and proliferation cellular nuclear antigen(PCNA) were determined. Results:RA exhibited anti-proliferative effects on cultured HSCs in a time and concentration dependent manner showing an IC50 of 276 μg/mL and 171 μg/mL for 24 h and 48 h,respectively,with morphological reversion of activated stellate cell morphology to quiescent form. It significantly improved ALT,AST,oxidative stress markers and reduced TIMP-1,HP levels,inflammatory markers and fibrosis score(S1 vs S4). Furthermore,reduction in α-SMA plus elevation in caspase-3 expressions of HSCs in vitro and in vivo associated with an inhibition in proliferation of damaged hepatocytes were recorded. Conclusions:RA impeded the progression of liver fibrosis through inhibition of HSCs activation/proliferation and induction of apoptosis with preservation of hepatic architecture.

Anti-fibrotic effect of rosmarinic acid on inhibition of pterygium epithelial cells

AIM： To investigate the anti-fibrosis effect of rosmarinic acid（RA） in pterygium epithelial cells（PECs） to determine if RA is a potent agent for treating pterygium.METHODS： The PECs（1×10-4 cells/mL） were treated with 100 μmol/L of RA for 1, 3 and 6h. After RA treatment, the cell viability was determined by staining with acridine orange/DAPI and analysis via a NucleoC ounter NC-3000. The protein expression levels of type I collagen, transforming growth factor beta-1（TGF-β1）, TGF-β type Ⅱ receptor（TGF-βRⅡ）, p-Smad1/5, p-Smad2, p-Smad3, and Smad4 of the cell lysates were measured by Western blot analysis.RESULTS： The cell viability of PECs was significantly decreased after RA treatment（P〈0.01）. As the result, RA reduced the protein expression of typeⅠcollagen and TGF-β1 of PECs. Additionally, RA also inhibited TGF-β1/Smad signaling by decreasing the protein expressions of TGF-βRII, p-Smad1/5, p-Smad2, p-Smad3, and Smad4.CONCLUSION： This study demonstrate that RA could inhibit fibrosis of PECs by down-regulating type I collagen expression and TGF-β1/Smad signaling. Therefore, RA is a potent therapeutic agent for the treatment of pterygium.

Synergistic effect of rosmarinic acid with antibiotics against Staphylococcus aureus and MRSA

OBJECTIVE To evaluate the synergistic effect of rosmarinic acid(RA)with antibiotics against Staphylococcus aureus(S.aureus)and MRSA and to identify the possible mechanism of action responsible for synergism if any.METHODS The antibacterial activity of Rosmarinic acid was studied for its zone of inhibition by agar well diffusion and MIC determination by liquid broth dilution technique against MRSA and VRSA.The synergistic effect of RA with antibiotics like amoxicillin,ofloxacin and vancomycin was evaluated by Broth checker board method and further confirmed with time kill kinetic studies.Microbial Surface Components Recognizing Adhesive Matrix Molecules(MSCRAMMs)were isolated by protein precipitation technique and its expression was studied by SDS PAGE.Further RA was evaluated for its beta lactamase inhibition property.RESULTS Rosmarinic acid exhibited antibacterial activity against S.aureus and MRSAby showing a MIC value of 0.8mg·mL-1 against S.aureus and 10mg·mL-1 against MRSA.Rosmarinic acid at 1/4X MIC value reduced the MIC of vancomycin,amoxicillin and ofloxacin by 1/4 times against S.aureus.But against MRSA,vancomycin was found to be synergistic.All the synergistic combinations have shown FIC value of 0.5.In order to measure the kinetics of the anti-bacterial activity,the bacterial growth rate with RA,antibiotics and synergistic combinations against S.aureus and MRSA was studied.It is observed that the synergistic combinations showed better time kill kinetics as compared to RA and antibiotics.Further RA could able to destruct the cell surface proteins MSCRAMMs which was studied by SDS PAGE.RA was also found to showβlactamase inhibiting property.CONCLUSION It is concluded that RA possess antibacterial activity but at a very higher concentration(in mg/mL)against S.aureus and MRSA.However it shows synergism with antibiotics and could able to reduce the MIC of antibiotics.Thus RA could be developed as an adjuvant for antibiotics against S.aureus and MRSA caused infections.Further studies are needed to identify the mechanism for its synergism with antibiotics.

Interactions of acetylcholinesterase with salvianolic acid B and rosmarinic acid from Salvia miltiorhiza water extract investigated by NMR relaxation rate

In order to understand whether the ameliorating effect on old ages memory disorder by the root of Salvia miltiorhiza is related to the acetylcholinesterase （ACHE） inhibition, two main ingredients, salvianolic acid B （1） and rosmarinic acid （2）, which were isolated from S. miltiorhiza water extract, were investigated in vitro by NMR relaxation rate in this work. The results showed that the proton selective relaxation rates and the molecular rotational correlation time of proton pairs for compounds 1 and 2 increased significantly by adding of AChE in mixing solution. The study reveals that the two compounds might bind to the enzyme and have ACHE inhibitory effect, which could contribute to the ameliorating effect at some extent on old ages memory disorder.

Inhibition Effects of Salvianolic Acid M and Rosmarinic Acid in Salvia deserta Schang on Human Aldose Reductase

[Objectives]This study aimed to investigate the inhibition effects of two chemical constituents in Salvia deserta Schang on human aldose reductase( AR). [Methods]The isolated salvianolic acid M and rosmarinic acid were prepared into different concentrations of solutions for in vitro enzyme kinetics experiments. [Results]Salvianolic acid M and rosmarinic acid had the strongest inhibition effects at the concentration of 1 g/L. [Conclusions]Salvianolic acid M and rosmarinic acid have inhibition effects on human aldose reductase. The IC50 of salvianolic acid M is 0. 79 μg/mL,and that of rosmarinic acid is 1. 13 μg/mL.

Total Synthesis of (±)-Butyl Ester of Rosmarinic Acid

±)-Butyl ester of rosmarinic acid 1 was synthesized by 5-step reactions through the two key intermediates 2 and 3, the total yield was 23.9% and 25.1%, respectively.

Chemical Fingerprint and Anti-Sickling Activity of Rosmarinic Acid and Methanolic Extracts from Three Species of Ocimum from DR Congo

The aim of this study was to characterize the polyphenolic composition by determination of chemical fingerprints of Methanolic extracts of Ocimum canum Sims, Ocimum basilicum L. and Ocimum gratissimum L. from Democratic Republic of Congo and to compare their antisickling activity of that of rosmarinic acid, the major compound to those of methanolic extracts. Phytochemical analysis performed by TLC and HPLC analysis, showed that rosmarinic acid is the most abundant phenolic acid in these Ocimum species according to the following order O. basilicum L., O. gratissimum L. and O. canum Sims. Methanolic extracts of these three species and pure rosmarinic acid showed significant antisickling activities with minimal concentration of normalization values of 0.18 ± 0.03, 0.23 ± 0.04, 0.26 ± 0.04 and 0.31 ± 0.05 mg/mL for rosmarinic acid, O. basilicum L., O. gratissimum L. and O. canum Sims methanolic extracts respectively. The antisickling activity order is the same as that of the rosmarinic acid content, indicating that this polyphenolic acid would be among the main active molecules in these extracts.

Investigation of the Effect of Rosmarinic Acid on Cyclophosphamide-Induced Gonadal Toxicity

The aim of this study was to investigate the effect of rosmarinic acid against gonadal toxicity caused by cyclophosphamide, an important anticancer drug. A total of 28 rats were divided into 4 groups, with 7 animals in each group. The groups were created as follows;Group 1 (control) (n = 7): Subjects received only 1 ml of 0.9% saline solution per day intraperitoneally for 14 days. Group 2 (Rosmarinic Acid) (n = 7): The subjects were given 20 mg/kg Rosmarinic acid intraperitoneally for 14 days. Group 3 (Cyclophosphamide) (n = 7): Only 1 ml of 0.9% saline solution was administered intraperitoneally to the subjects for the first 7 days. Cyclophosphamide 20 mg/kg per day was administered intraperitoneally for the last 7 days (from the 8th day). Group 4 (Rosmarinic Acid + Cyclophosphamide) (n = 7): The subjects were given 20 mg/kg Rosmarinic acid daily intraperitoneally for 14 days. From the 8th day of the experiment to the end of the experiment (last 7 days of the experiment), 20 mg/kg cyclophosphamide was given intraperitoneally daily. At the end of the experiment, body weights of all rats were measured first. Afterwards, the weights of testicular tissue samples were measured and the averages of the weights were taken. Routine tissue follows up was performed on the testicular tissues taken. Hematoxylin-eosin staining was applied to tissue sections of 5 μm thickness. As a result of the statistical analysis, it was determined that cyclophosmadine decreased body and testicular weight, but rosmarinic acid had a protective effect on the contrary. It was concluded that cyclophosmadine caused damage to the basement membrane structure, Sertoli, Leydig and germ cells, but these structures were preserved due to the protective effect of rosmarinic acid. Despite the toxic effect of CP, rosmarinic acid is thought to have a significant curative effect on the spermatogenetic process and seminiferous tubule structure in the gonads.

Single-cell transcriptomics reveals the ameliorative effect of rosmarinic acid on diabetic nephropathy-induced kidney injury by modulating oxidative stress and inflammation

Diabetic nephropathy(DN)is a severe complication of diabetes,characterized by changes in kidney structure and function.The natural product rosmarinic acid(RA)has demonstrated therapeutic effects,including anti-inflammation and anti-oxidative-stress,in renal damage or dysfunction.In this study,we characterized the heterogeneity of the cellular response in kidneys to DN-induced injury and RA treatment at single cell levels.Our results demonstrated that RA significantly alleviated renal tubular epithelial injury,particularly in the proximal tubular S1 segment and on glomerular epithelial cells known as podocytes,while attenuating the inflammatory response of macrophages,oxidative stress,and cytotox-icity of natural killer cells.These findings provide a comprehensive understanding of the mechanisms by which RA alleviates kidney damage,oxidative stress,and inflammation,offering valuable guidance for the clinical application of RA in the treatment of DN.

An injectable conductive hydrogel with dual responsive release of rosmarinic acid improves cardiac function and promotes repair after myocardial infarction

Myocardial infarction (MI) causes irreversible damage to the heart muscle, seriously threatening the lives of patients. Injectable hydrogels have attracted extensive attention in the treatment of MI. By promoting the coupling of mechanical and electrical signals between cardiomyocytes, combined with synergistic therapeutic strategies targeting the pathological processes of inflammation, proliferation, and fibrotic remodeling after MI, it is expected to improve the therapeutic effect. In this study, a pH/ROS dual-responsive injectable hydrogel was developed by modifying xanthan gum and gelatin with reversible imine bond and boronic ester bond double crosslinking. By encapsulating polydopamine-rosmarinic acid nanoparticles to achieve on-demand drug release in response to the microenvironment of MI, thereby exerting anti-inflammatory, anti-apoptotic, and anti-fibrosis effects. By adding conductive composites to improve the conductivity and mechanical strength of the hydrogel, restore electrical signal transmission in the infarct area, promote synchronous contraction of cardiomyocytes, avoid induced arrhythmias, and induce angiogenesis. Furthermore, the multifunctional hydrogel promoted the expression of cardiac-specific markers to restore cardiac function after MI. The in vivo and in vitro results demonstrate the effectiveness of this synergistic comprehensive treatment strategy in MI treatment, showing great application potential to promote the repair of infarcted hearts.

Novel colorimetric,photothermal and up-conversion fluorescence triple-signal sensor for rosmarinic acid detection

Rosmarinic acid(RA)is promising as a natural and nontoxic food additive.However,many analysis methods for RA generally depend on large instruments and single signals for quantitative detection.A new upconversion fluorescence,colorimetric and photothermal multi-modal sensing strategy is developed for the quantification of RA.β-cyclodextrin(CD)modified citric acid(Cit)wrapped NaYF_(4):Yb/Er-Cit-CD(Y:Yb/Er-Cit-CD)up-conversion nanocomposite has been synthesized,which emits green fluorescence at 550nm under 980nm near-infrared(NIR)excitation.In the presence of oxidized 3,3',5,5'-tetramethylbenzidine(oxTMB),the green fluorescence is significantly quenched attributed to the fluorescence inner filter effect(IFE)between oxTMB and Y:Yb/Er-Cit-CD.When RA is intervened,blue oxTMB is reduced to colorless 3,3',5,5'-tetramethylbenzidine(TMB)inducing the recovery of up-conversion fluorescence.At the same time,colorimetric and photothermal signals readout can be easily achieved thanks to the color indication and photothermal effect of the oxTMB.The constructed Y:Yb/Er-Cit-CD/oxTMB sensor displays high sensitivity,visibility and simplicity for RA,and the limits of detection(LOD)for fluorescence,colorimetric and photothermal were 0.004μmol/L,0.036μmol/L and 0.043μmol/L,respectively.This sensing system is successfully performed for the detection of RA in food samples.

A New Homogenizing Technology to Obtain Rosmarinic Acid from Perilla Oil Meal

Objective To optimize the extraction technology of the active component, rosmarinic acid, an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid, in perilla oil meal for the first time by a new homogenizing technology called smashing tissue extraction (STE). Methods Orthogonal design was used to optimize the extraction condition. The content of rosmarinic acid was quantified from the methanol crude extract with the help of HPLC. Results The optimization of STE process to get rosmarinic acid from the perilla oil meal was the ratio of liquid to solid material at 10:1 and the power of extraction at 150 V, extracting twice (2 min for each time). Conclusion STE could be applied to extracting the active ingredients from the oil meals due to its high extraction efficiency. This new homogenizing technology has advantages on saving extraction time, raising extraction efficiency, and maintaining the temperature sensitive constituents.

Effects of Rosmarinic Acid on DNA Integrity and H19 Differentially Methylated Region Methylation Levels in Human Sperm Preserved by Freeze-Drying

Original Article Effects of Rosmarinic Acid on DNA Integrity and H19 Differentially Methylated Region Methylation Levels in Human Sperm Preserved by Freeze-Drying Wang Yi-Yu,Zhu Wei-Jie Published 2021-03-25 Cite as Reprod Dev Med,2021,05 Reprod Dev Med,2021,05(1):9-14.DOI:10.4103/2096-2924.309790 Abstract Objective:To investigate the effects of rosmarinic acid(RA)on the DNA integrity and methylation levels of the H19 differentially methylated region(DMR)of freeze-dried human sperm after 1 week and 6 months of storage at 4℃.Methods:Semen samples from 15 healthy normospermic donors were used in this study.The samples were divided into five groups,including the control group with fresh sperm and four experimental groups with freeze-dried sperm(1-week storage with EGTA buffer solution,Group A;1-week storage with EGTA buffer solution containing 105μmol/L RA,Group B;6-month storage with EGTA buffer solution,Group C;and 6-month storage with EGTA buffer solution containing 105μmol/L RA,Group D).DNA integrity was evaluated using the sperm chromatin dispersion test.H19 DMR methylation levels were detected by bisulfite sequencing polymerase chain reaction.Results:After 1 week of storage,no differences in sperm DNA integrity were observed among Groups A,B,and controls(P>0.05).After 6 months of storage,the sperm DNA integrity of Group D did not change significantly compared with that of the control group(P>0.05),whereas that of Group C decreased significantly(P<0.05).There were no differences in H19 DMR methylation levels among the five groups(P>0.05).Conclusions:The DNA integrity of freeze-dried human sperm can be effectively protected by adding RA within 6 months,and the H19 DMR methylation level of human sperm can be maintained for 6 months after freeze-drying.

Algerian mint species:high performance thin layer chromatography quantitative determination of rosmarinic acid and in vitro inhibitory effects on linoleic acid peroxidation

Objective:To determine the quantitative paremeters of rosmarinic acid in Algerian mints,Mentha spicata L.(M.spicata),Mentha pulegium L.and Mentha rotundifolia(L.)Huds by high performance thin layer chromatography(HPTLC)-densitometric method and screen the effects of these plant extracts on linoleic acid peroxidation.Methods:The analyses were performed on HPTLC silica gel 60 F254 plates with chloroform:acetone:formic acid(75:16.5:8.5,v/v)as the mobile phase.Rosmarinic acid was determined in UV at 365 nm and fluorescence at λ_(exc)325 nm with a 550 nm filter,respectively.The effects of plants extracts on linoleic acid peroxidation were measured by an indirect in vitro colorimetric method.Results:Chromatographic resolution permitted reliable quantification in both measurement modes and calibration curves were linear in a range of 150-1000 ng/spot.M.spicata was found to contain significantly higher concentrations of rosmarinic acid.The densitometric quantification allowed the analysis of many samples in a short time with reasonable precision(total precision for Mentha spp extracts,5.1%and 5.8%for UV and fluorescence detection,respectively).The HPTLC data,allied to assays of linoleic acid peroxidation prevention,suggested the potential of M.spicata(52%Trolox®equivalents)as a natural source for inhibitors of lipid peroxidation.Conclusions:Densitometry can be used for routine determination and quality control of rosmarinic acid in herbal and formulations containing Mentha species.

The Effect of <i>Prunella</i>on Anti-Inflammatory Activity in RAW264.7 Mouse Macrophage Cells

The extracts of Prunella vulgaris L. (Labiatae), a popular Western and Chinese herbal medicine, was shown to have anti-inflammatory properties, which might be due to partially, their rosmarinic acid content. Inhition of prostaglandine E2 (PGE2) production in lipopolysaccharide (LPS) stimulated RAW264.7 mouse macrophage cells was assessed with an enzyme immunoassay (EIA) following 8-hour treatments with Prunella vulgaris extracts or fractions. Results showed that 95% ethanol extracts from P. vulgaris significantly inhibited PGE2 production. In further studies, fraction 2 from the 95% ethanol extract of P. vulgaris significantly reduced PGE2 production at 66 μg/ml (72% reduction). Cytotoxic-ity did not play a role in the noted reduction of PGE2 seen in either the extracts or fractions from P. vulgaris. High performance liquid chromatography analysis showed that there was 1.4 mM rosmarinic acid in 95% ethanol Prunella extract (201 mg/ml crude extract). Our results suggest that rosmarinic acid may contribute toward the anti-inflammatory activity of Prunella in a dose-response manner. Prunella might have a potential to be used as a functional food for anti-inflammatory activity.