Study on quality standard for Miao medicine Clinopodium gracile / 中国药房

OBJECTIVE To establish the quality standard of Clinopodium gracile. METHODS Ten batches of C. gracile were collected to perform appearance and property identification， microscopic identification and thin layer chromatography （TLC） identification. Moisture， total ash， acid-insoluble ash and dilute ethanol extract were detected， and the content of rosmarinic acid was determined by HPLC. RESULTS The stem of C. gracile was slender， square columnar， covered by white fluff， the surface was grayish green or greenish brown； epidermal cells， non-glandular hairs， cortical cells and so on were seen in the cross section of the stem. Non-glandular hairs， ducts， wood fibers， mesophyll cells and so on could be seen in the powder. Results of TLC identification showed that there were spots of the same color in the chromatographic position corresponding to the chromatographic position of buddlejasaponin Ⅳb control. The contents of water， total ash， acid-insoluble ash， dilute ethanol extract and rosmarinic acid in 10 batches of samples were 8.69%-12.33%， 5.96%-13.33%， 0.14%-3.29%， 18.57%-32.61%， 0.35%-0.82%， respectively. The average values were 10.10%， 9.73%， 1.06%， 23.54% and 0.56%， respectively. CONCLUSIONS The established method can be used for quality control of C. gracile. It is preliminarily proposed that the ash content in the herb should not exceed 12.0%， the total ash content should not exceed 12.0%， the acid-insoluble ash content should not exceed 1.5%， the dilute ethanol extract should not be less than 18.0%， and the rosmarinic acid content should not be less than 0.45%.

Applicability of [18F]FDG/PET for investigating rosmarinic acid preconditioning efficacy in a global stroke model in mice

ABSTRACT Positron emission tomography (PET) is a non-invasive nuclear imaging technique that uses radiotracers to track cell activity. The radiopharmaceutical 18F-fluoro-2-deoxyglucose ([18F] FDG) is most commonly used in nuclear medicine for the diagnosis of various diseases, including stroke. A stroke is a serious condition with high mortality and morbidity rates. Rosmarinic acid (RA) is a promising therapeutic agent that exerts neuroprotective effects against various neurological diseases. Therefore, this study aimed to evaluate the applicability of [18F]FDG/PET for investigating the neuroprotective effects of RA in case of a global stroke model in mice. The [18F]FDG/PET technique facilitates the observation of ischemia and reperfusion injuries in the brain. Moreover, the recovery of glucose metabolism in three specific brain regions, the striatum, superior colliculus, and inferior colliculus, was observed after preconditioning with RA. It was concluded that the [18F]FDG/PET technique may be useful for stroke diagnosis and the assessment of treatment response. In addition, a long-term longitudinal study using biochemical analysis in conjunction with functional imaging may provide further conclusive results regarding the effect of RA on cerebral ischemia.

Optimization of extraction process of Sarcandra glabra by combination of entropy weight method with Box-Behnken response surface method / 中国药房

OBJECTIVE To optimize the extraction process of Sarcandra glabra. METHODS The contents of rosmarinic acid and isofraxidin in S. glabra were determined by HPLC； ultrasonic time， ultrasonic temperature， solid-liquid ratio （mL/g） and methanol volume fraction were investigated by single factor test. Based on the results of single factor test， experimental scheme was designed by Box-Behnken response surface method， and the entropy weight method was used to assign the weight of each index and calculate the comprehensive score. Taking the comprehensive score as the evaluation index， the extraction process of S. glabra was optimized， and then optimized extraction process was verified. RESULTS The optimal extraction technology of S. glabra included ultrasonic time of 40 min， ultrasonic temperature of 45 ℃， liquid-solid ratio of 50∶1， methanol volume fraction of 70%. The results of 3 times of verification experiment showed that average comprehensive score was 0.988 6， and the RSD was 0.50%. The deviation between the actual value and the predicted value （0.985 1） of each comprehensive score was within ±1%. CONCLUSIONS The optimized extraction method is stable， feasible and repeatable， which can provide reference for extraction of S. glabra.

SmHPPR1 from Salvia miltiorrhiza regulated the biosynthesis of salvianolic acids / 药学学报

italic>Salvia miltiorrhiza Bunge is a traditional Chinese medicinal herb widely used to treat cardiovascular and cerebrovascular diseases at clinic. Its main water-soluble components are rosmarinic acid (RA) and salvianolic acid B (SAB), which are produced by phenylpropanoid pathway. 4-Hydroxyphenylpyruvate reductase (HPPR) is a key enzyme in phenylpropanoid metabolism pathway. SmHPPR1 was cloned from S. miltiorrhiza and was constructed into plant expression vector pJR-SmHPPR1. On this basis, SmHPPR1 transgenic Arabidopsis plants were induced and the content of 4-hydroxyphenyllactic acid (pHPL) was determined. SmHPPR1-overexpressing (SmHPPR1-OE) hairy roots of S. miltiorrhiza were obtained and the concentration of active components and transcriptome analysis were performed. The results showed that the concentration of pHPL in SmHPPR1 transgenic Arabidopsis T1 was 0.594 mg·g-1 dry weight. The concentration of RA, SAB and total salvianolic acid in SmHPPR1-OE-3 hairy roots were 1.09, 1.29, 1.15 times of that in control-3, respectively, and the content of Danshensu was 36.26% of that in control-3. Transcriptomic analysis revealed that overexpression of SmHPPR1 caused the upregulation of other phenylpropanoid pathway genes like SmTAT2. Protein-protein interaction indicated CYT (TR74706_c0_g1), NADP+ (TR26565_c0_g1) and NADP+ (TR68771_c0_g1) is the central node of the network and participated in metabolic process and cellular process. The tracking work in this study proved that SmHPPR1 could catalyze the reduction of 4-hydroxyphenylpyruvic acid to 4-hydroxyphenyllactic acid in SmHPPR1 transgenic Arabidopsis, and SmHPPR1-overexpressing in hairy roots of S. miltiorrhiza could increase the concentration of salvianolic acids through synergistically regulating other pathway genes.

Ameliorative impacts of floral extract of Salvia species on oxidative stress and inflammation in rats renal ischemia/reperfusion

I/R injury is a potentially serious problem that is encountered during a variety of medical and surgical procedures, such as thrombolytic therapy, organ transplantation and coronary angioplasty, The basic pathophysiology of I/R injury is microvascular dysfunction which is developed following reperfusion of ischemic tissues. It has clinical importance because of its frequent occurrence and mortality in some surgical conditions such as renal transplantation. Here, we investigated the protective effect of Salvia extracts on kidneys against I/R injury. Forty Spraque Dawley rats were divided into 5 groups. Right nephrectomy was performed to all groups. Gr. I, control; Gr. II, I/R; Gr. III & IV, I/R+50 and I/R+100 mg/kg Salvia floral extract; and Gr. V with I/R+50 mg/kg Rosmarinic acid. Salvia and Rosmarinic acid for 7 days was given single dose as a gavage.60 min ischemia, 60 min reperfusion were applied to groups except control. Intracardiac blood samples were taken, Blood urea nitrogen, creatine, malondialdehyde, myeloperoxidase, nitric oxide and chitotriosidase levels were detected. Mean values were evaluated by statistical analysis. The renal tissues were examined under light microscopy. Based on our biochemical and histological data, Salvia floral extract has potent anti-inflammatory and antioxidant effects against renal structure and function.

Cognition enhancing effect of rosemary (Rosmarinus officinalis L.) in lab animal studies: a systematic review and meta-analysis

Patients with mild cognitive impairment eventually progress to Alzheimer's disease (AD) causing a strong impact on public health. Rosmarinus officinalis has long been known as the herb of remembrance and can be a potential cognition enhancer for AD. The aim of this review was to summarize the qualitative and quantitative aspects of R. officinalis and its active constituents in enhancing cognition. A structured search was conducted on Google Scholar and PubMed to find relevant studies that assessed the effect of R. officinalis extract or any of its active constituents on cognitive performance in animals. The following information was extracted from each study: 1) article information; 2) characteristics of study animals; 3) type of intervention: type, dose, duration, and frequency of administration of R. officinalis; and 4) type of outcome measure. Data were analyzed using Review Manager and meta-analysis was performed by computing the standardized mean difference. Twenty-three studies were selected for qualitative analysis and fifteen for meta-analysis. From the fifteen included papers, 22 with 35 comparisons were meta-analyzed. Effect sizes for intact and cognitively impaired animals were 1.19 (0.74, 1.64) and 0.57 (0.19, 0.96), indicating a positive effect on both groups. The subgroup analyses showed substantial unexplained heterogeneity among studies. Overall, R. officinalis improved cognitive outcomes in normal and impaired animals, and results were robust across species, type of extract, treatment duration, and type of memory. However, studies had a considerable amount of heterogeneity, and subgroup analyses failed to find any heterogeneity moderator.

Quality study of Prunellae spica dispensing granules based on standard decoction / 国际中医中药杂志

Objective:To establish the quality evaluation method of Prunellae spica dispensing granules based on three quality indexes of standard decoction. Methods:Fourteen batches of Prunellae spica were collected from different habitats. According to technical requirements, fourteen batches of Prunellae spica standard decoction and three batches of formula granules were prepared and the paste-forming rates were calculated. The fingerprints of Prunellae spica standard decoction and formula granules were established by Ultra High Performance Liquid Chromatography (UPLC). The similarity values of fingerprints between dispensing granules and standard decoction were calculated. The content and transferring rate of Rosmarinic acid were determined and calculated. Results:The average paste-forming rate of Prunellae spica was (12.59±2.32)%. The paste-forming rates of the three batches were 11.14%, 10.78% and 10.39% respectively. The average content of Rosmarinic acid in standard decoction was (18.99±9.74)mg/g. The average transferring rate was (60.58±7.87)%. The contents of three batches were 7.40 mg/g, 7.49 mg/g and 7.09 mg/g. The transferring rates were 52.06%, 50.10% and 50.40% respectively. Nine common fingerprint peaks were identified in the fingerprints of standard decoction and formula granules, two of which were identified as Rosmarinic acid and Caffeic acid by comparison of reference substance. The fingerprints similarity of Prunellae spica dispensing granules and standard decoction were 0.954, 0.973 and 0.952, respectively. Conclusions:The quality indexes of three batches of formulation granules are consistent with standard decoction. This method could provide reference for the establishment of quality standard of Prunellae spica dispensing granules.

Inhibitory effect of rosmarinic acid on high glucose-induced angiogenesis of human retinal microvascular endothelial cells and the activation of NLRP3 inflammasome pathway / 中华实验眼科杂志

Objective:To investigate the inhibitory effects of rosmarinic acid (RA) on high glucose-induced angiogenesis of human retinal microvascular endothelial cells (HRMEC) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome pathway-related proteins.Methods:The HRMEC were divided into control group, high glucose group, high glucose+ low concentration RA group, high glucose+ medium concentration RA group, and high glucose+ high concentration RA group, and were cultured in vitro with conventional medium, 30 mmol/L D-glucose medium, 30 mmol/L D-glucose+ 25 μmol/L RA medium, 30 mmol/L D-glucose+ 50 μmol/L RA medium and 30 mmol/L D-glucose+ 100 μmol/L RA medium accordingly.The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to detect the cell proliferation.Transwell assay was performed to detect the cell migration.Matrigel assay was employed to determine the tube formation ability of cells.Western blot was utilized to detect the expression levels of NLRP3, apoptosis-associated speck like protein (ASC) and cysteinyl aspartate-specific protease-1 (Caspase-1). Enzyme-linked immunosorbent assay (ELISA) kit was used to detect the concentrations of interleukin (IL)-1β and IL-18 in supernatant of cell culture. Results:The cell proliferation rate, the number of migrated cells and the number of formed tubes were (100.00±0.92)%, 37.67±9.02 and 45.00±4.58 in the control group, (163.56±1.46)%, 117.33±7.23 and 95.00±9.54 in the high glucose group, (152.29±2.90)%, 78.67±4.04 and 84.67±1.53 in the high glucose+ low concentration RA group, (147.72±2.22)%, 65.33±4.16 and 71.00±3.61 in the high glucose+ medium concentration RA group, (132.47±0.74)%, 52.67±6.81 and 60.00±1.00 in the high glucose+ high concentration RA group, respectively.There were statistically significant differences in cell proliferation rate, the number of migrated cells and formed tubes among all groups ( F=537.07, 64.63, 45.58; all at P<0.001). Compared with the control group, the cell proliferation rate, the number of migrated cells and formed tubes were significantly increased in the high glucose group, high glucose+ low concentration RA group, high glucose+ medium concentration RA group and high glucose+ high concentration RA group, showing statistical significances (all at P<0.05). Compared with the high glucose group, the cell proliferation rate, the number of migrated cells and formed tubes were significantly decreased in the different concentrations RA groups (all at P<0.05). With the increase of RA concentration, the cell proliferation rate, the number of migrated cells and formed tubes were decreased, and there were statistical differences among high glucose+ low/medium/high concentrations RA groups (all at P<0.05). There were significantly differences in the relative expression levels of NLRP3, ASC and Caspase-1 proteins in cells and the concentrations of IL-1β and IL-18 in cell culture supernatant among all the five groups ( F=145.12, 422.82, 463.79, 2 019.96, 33 406.97; all at P<0.001). Compared with the control group, the relative expression levels of NLRP3, ASC and Caspase-1 proteins as well as the concentrations of IL-1β and IL-18 in cell culture supernatant were significantly increased in the high glucose group, high glucose+ low concentration RA group, high glucose+ medium concentration RA group and high glucose+ high concentration RA group (all at P<0.05). Compared with the high glucose group, the expression levels of NLRP3, ASC and Caspase-1 proteins as well as the concentrations of IL-1β and IL-18 were decreased in the different concentrations RA groups, and the differences were statistically significant (all at P<0.05). With the increase of RA concentration, the expression levels of NLRP3, ASC and Caspase-1 proteins as well as the concentrations of IL-1β and IL-18 were decreased, and there were statistically significant differences among high glucose+ low/medium/high concentrations RA groups (all at P<0.05). Conclusions:RA can inhibit proliferation, migration and tube formation of HRMEC induced by high glucose, and inhibit high glucose-induced activation of NLRP3 inflammasome signaling pathway.

Quality standard of Perillae Folium based on multicomponent determination with HPLC method / 中国中药杂志

This research established the HPLC methods for the determination of perillaketone, perillaldehyde, caffeic acid, scutellarin, and rosmarinic acid in 33 batches of Perillae Folium. Kromasil C_(18)(4.6 × 250 mm, 5 μm) chromatographic column was used, and the mobile phase for determination of the perillaketone and perillaldehyde was methanol-water(55∶45) solution, at a flow rate of 1.0 mL·min~(-1), with the column temperature at 30 ℃. The mobile phase for the determination of caffeic acid, scutellarin and rosmarinic acid was methanol(A)-0.2% phosphoric acid aqueous solution(B) with gradient elution(0-20 min, 25%-30% A; 20-60 min, 30%-43% A). The flow rate was 1.0 mL·min~(-1) and the column temperature was set at 30 ℃. The results showed that the established method can achieve good separation of the five components in samples, with a good linear relationship and high accuracy, indicating that the methods can be used for the determination of Perillae Folium. The results showed that all samples contained five components. And the content of rosmarinic acid(0.04%-1.57%) > scutellarin(0.03%-0.77%) > perillaldehyde(0.02%-0.66%) > perillaketone(0.03%-0.30%) > caffeic acid(0.006%-0.07%). Thirty-three Batches of Perillae Folium can be grouped into 5 categories. There are certain content rules and region specificities under different clusters. Perillaketone, perillaldehyde, and rosmarinic acid can be used as the main markers to evaluate the quality of Perillae Folium.

Cloning and functional analysis of caffeic acid and rosmarinic acid glycosyltransferases from Arnebia euchroma / 中国中药杂志

Caffeic acid and its oligomers are the main water-soluble active constituents of the traditional Chinese medicine(TCM) Arnebiae Radix. These compounds possess multiple biological activities such as antimicrobial, antioxidant, cardiovascular protective, liver protective, anti-liver fibrosis, antiviral and anticancer activities. The phenylpropanoid pathway in plants is responsible for the biosynthesis of caffeic acid and its oligomers. Glycosylation can change phenylpropanoid solubility, stability and toxic potential, as well as influencing compartmentalization and biological activity. In view of the important role played by de-glycosylation in the regulation of phenylpropanoid homeostasis, the biosynthesis of caffeic acid and its oligomers are supposed to be under the control of relative UDP-glycosyltransferases(UGTs). Through the data mining of Arnebia euchroma transcriptome, we cloned 15 full-length putative UGT genes. After recombinant expression using the prokaryotic system, the crude enzyme solution of the putative UGTs was examined for the glycosylation activities towards caffeic acid and rosmarinic acid in vitro. AeUGT_01, AeUGT_02, AeUGT_03, AeUGT_04 and AeUGT_10 were able to glycosylate caffeic acid and/or rosmarinic acid resulting in different mono-and/or di-glycosylated products in the UPLC-MS analyses. The characterized UGTs were distantly related to each other and divided into different clades of the phylogenetic tree. Based on the observation that each characterized UGT exhibited substrate or catalytic similarity with the members in their own clade, we supposed the glycosylation abilities towards caffeic acid and/or rosmarinic acid were evolved independently in different clades. The identification of caffeic acid and rosmarinic acid UGTs from A. euchroma could lead to deeper understanding of the caffeic acid oligomers biosynthesis and its regulation. Furthermore, these UGTs might be used for regiospecific glycosylation of caffeic acid and rosmarinic acid to produce bioactive compounds for potential therapeutic applications.

Contents determination of eight phenolic compounds in Perilla frutescens leaves of different cultivation years and harvesting periods / 中国中药杂志

A method was established for content determination of two kinds of phenolic acids, including rosmarinic acid)(RA) and caffeic acid(CA), and six kinds of flavonoids including scutellarein-7-O-diglucuronide(SDG), luteolin-7-O-diglucuronide(LDG), apigenin-7-O-diglucuronide(ADG), scutellarin-7-O-glucuronide(SG), luteolin-7-O-glucuronide(LG), and apigenin-7-O-glucuronide(AG) in Perilla frutescens leaves. The content of eight chemical components was measured based on ten P. frutescens germplasms of different chemotypes of volatile oil, different cultivated years, and different harvesting periods. The results showed that there was a great difference between the two kinds of constituents of different germplasms. The total content of the two phenolic acids was 2.24-34.44 mg·g~(-1), and the total content of the six flavonoids was 11.55-34.71 mg·g~(-1). Then according to content from most to least, the order of each component was RA(2.13-33.97 mg·g~(-1)), LDG(1.31-14.80 mg·g~(-1)), SG(1.97-8.45 mg·g~(-1)), ADG(2.68-7.60 mg·g~(-1)), SDG(1.16-5.87 mg·g~(-1)), LG(0.78-1.91 mg·g~(-1)), AG(0.56-1.00 mg·g~(-1)), and CA(0.11-0.68 mg·g~(-1)). The chemical contents of the 5 PA-type germplasms in 2017 were mostly higher than those in 2018 showing a large variation with the cultivation years. These contents of two kinds of phenolic acids of 9 germplasms fluctuated with the harvesting time. The content decreased before early flower spike(the 3~(rd) to 18~(th) in August) at first and began to increase in flowering and fruiting period(the 18~(th) in August to 2~(nd) in September). However, these contents had slowly decreasing trend after 2~(nd) in September till 17~(th) in the same month. Interestingly, the content raised again in the maturity of fruits. The variation tendency of contents in six kinds of flavonoids components was inconsistent in different germplasms with the variation of harvesting time. The content of flavonoids in part of germplasms was negatively correlated with the fluctuation of phenolic acids. There was no correlation between phenolic acids and chemical type of the volatile oil. This paper may provide a reference for the high-quality germplasm of P. frutescens cultivation.

Anti-fibrotic effects of rosmarinic acid on Tenon's capsule fibroblasts stimulated with TGF-ß: therapeutic potential in ocular surgery / Efeito antifibrótico do Ácido Rosmarínico sobre os fibroblastos da Cápsula de Tenon estimulados com TGF-beta2: potencial terapêutico na cirurgia ocular

ABSTRACT Purpose: Collagen deposition and myofibroblast differentiation are critical factors related to excessive scarring in ocular surgeries. This study evaluated the anti-fibrotic activity of rosmarinic acid on rabbit Tenon's capsule fibroblasts stimulated with transforming growth factor- β2. Methods: Primary cultures of rabbit Tenon's capsule fibroblasts were treated with various concentrations of rosmarinic acid for 12 h, in the presence and absence of transforming growth factor-β2. After 48 h, the proliferation index of rabbit Tenon's capsule fibroblasts and the differentiation of myofibroblasts were investigated through immunofluorescence staining for proliferating cell nuclear antigen and alpha smooth muscle actin. An automated cell counter and colorimetric metabolic activity assay were used to evaluate cell number and viability. Collagen expression and production were determined by quantitative real-time polymerase chain reaction and hydroxyproline assay, respectively. Results: Unstimulated rabbit Tenon's capsule fibroblasts treated with any concentration of rosmarinic acid exhibited diminished collagen expression (p<0.01) but showed no differences in proliferation index. Transforming growth factor-β2 exposure induced myofibroblast differentiation and increased collagen production. Exposure to rosmarinic acid at 1.0 and 3.0 µM concentrations reduced the proliferation index (p<0.02), as well as the collagen expression and hydroxyproline content (p<0.05). Exposure to 3.0 µM rosmarinic acid reduced viability (p=0.035) in unstimulated rabbit Tenon's capsule fibroblasts and cell numbers (p=0.001) in both stimulated and unstimulated rabbit Tenon's capsule fibroblast cultures. Conclusions: Exposure to 1.0 µM rosmarinic acid was noncytotoxic and led to reduced collagen expression and proliferation of stimulated rabbit Tenon's capsule fibroblasts. These findings suggest that rosmarinic acid is a relatively non-injurious anti-fibrotic compound to rabbit Tenon's capsule fibroblasts, with potential application as an adjunctive agent in ocular procedures, particularly in glaucoma surgeries.

RESUMO Objetivo: A deposição de colágeno e a diferenciação de miofibroblastos são fatores chaves relacionados à cicatrização excessiva em cirurgias oculares. Este estudo avaliou a atividade anti-fibrótica do ácido rosmarínico nos fibroblastos da cápsula de Tenon de coelhos estimulados com o fator de crescimento transformador-β2. Métodos: Culturas primárias de fibroblastos da cápsula de Tenon de coelhos foram tratadas com várias concentrações de ácido rosmarínico por 12h, na presença e na ausência do fator de crescimento transformador-β2. Após 48h, o índice de proliferação dos fibroblastos da cápsula de Tenon de coelhos e a diferenciação dos miofibroblastos foram investigados por coloração por imunofluorescência para proliferação de antígeno nuclear celular e α-actina de músculo liso, respectivamente. Um contador automático de células e um ensaio de atividade metabólica colorimétrica foram utilizados para avaliar o número e a viabilidade das células. A expressão e produção do colágeno foram determinadas por reação quantitativa em cadeia da polimerase em tempo real e ensaio de hidroxiprolina, respectivamente. Resultados: Fibroblastos da cápsula de Tenon de coelhos não estimulados tratados com qualquer concentração de ácido rosmarínico exibiram diminuição de colágeno (p<0,01), mas não mostraram diferenças no índice de proliferação. A exposição ao fator de crescimento transformador-β2 induziu a diferenciação de miofibroblastos e aumentou a produção de colágeno. A exposição ao ácido rosmarínico nas concentrações de 1,0 e 3,0 µM reduziu o índice de proliferação (p<0,02), bem como a expressão de colágeno e a quantificação de hidroxiprolina (p<0.05). A exposição a 3,0 µM de ácido rosmarínico reduziu a viabilidade (p=0,035) de fibroblastos da cápsula de Tenon de coelhos não estimulados e o número de células (p=0,001) em culturas de fibroblastos da cápsula de Tenon de coelhos estimuladas e não estimuladas. Conclusões: A exposição ao ácido rosmarínico 1,0 µM foi não citotóxica e levou à expressão reduzida de colágeno e menor proliferação de fibroblastos da cápsula de Tenon estimulados pelo fator de crescimento transformador-β2. Esses achados sugerem que o ácido rosmarínico é um composto antifibrótico relativamente não lesivo aos fibroblastos da cápsula de Tenon de coelhos, com potencial aplicação como agente adjuvante em procedimentos oculares, particularmente em cirurgias de glaucoma.

Investigation of the role of rosmarinic acid treatment in regulating inflammation, cell damage, and angiogenesis in rat ovarian torsion and detorsion models

Abstract Purpose To investigate the protective effect of rosmarinic acid (RA) in ovarian ischemia/reperfusion injury using biochemical, histopathological, and immunohistochemical methods. Methods Wistar female rats (n = 32) were randomly divided into four groups: control, ischemia, ischemia-reperfusion, and ischemia-reperfusion with RA. Rosmarinic acid was given at a dose of 50 mg/kg by oral gavage three hours after reperfusion. Malondialdehyde (MDA) levels and glutathione peroxidase (GSH-Px) activities were determined in the ovary tissue homogenates for each rat. Results In the ischemia-reperfusion with RA group, the epithelial cells are regularly regulated at the periphery, and the degenerative changes in preantral and antral follicle cells are reduced. Follicle cells and cells in the corpus luteum showed a decrease in vascular endothelial growth factor (VEGF) expression, while VEGF demonstrated a positive reaction in vascular endothelial cells and stromal cells. The TNF-α expression due to the decreased degenerative effect and inflammation was positive in the macrophage cells. The expression of caspase-3 as an apoptosis change was negative in antral follicle cells and granular cells around the antral follicle. Conclusion Different doses of RA may be useful in preventing ischemic damage after vascularization, inflammation, and apoptotic development after ischemia/reperfusion.

The effect of rosmarinic acid on deformities occurring in brain tissue by craniectomy method. Histopathological evaluation of IBA-1 and GFAP expressions

Abstract Purpose To investigate the role of Rosmarinic acid (RA) in the prevention of traumatic brain injury and the immunohistochemical analysis of IBA-1 and GFAP expressions. Methods Healthy male rats were randomly divided into 3 groups consisting of 10 rats. Groups were as follows; control group, traumatic brain injury (TBI) group, and TBI+RA group. After traumatic brain injury, blood samples were taken from the animals and analyzed with various biochemical markers. And then IBA-1 and GFAP expressions were evaluated immunohistochemically. Results Significant results were obtained in all biochemical parameters between groups. Immunohistochemical sections showed IBA-1 not only in microglia and macrophage activity but also in degenerative neurons in blood vessel endothelial cells. However, GFAP reaction and post-traumatic rosmarinic acid administration showed positive expression in astrocytes with regular structure around the blood vessel. Conclusion Rosmarinic acid in blood vessel endothelial cells showed that preserving the integrity of astrocytic structure in the blood brain barrier may be an important antioxidant.

Assessment of the Antioxidative Potential of Rosmarinus officinalis L. (Lamiaceae) Irrigated with Static Magnetic Field-Treated Water

Abstract Phenolic compounds are one of the main groups of secondary metabolites in plants and are known for their antioxidant activity. Rosmarinus officinalis L. (rosemary) contains different phenolic compounds including carnosol, carnosic acid and rosmarinic acid. In Cuba, rosemary cultivation is limited because it is difficult to propagate and has a low yield. As a result, it was removed from the Herbal Medicine National Formulary. However, the National Public Health System has a strong interest in rosemary because of its value as a natural antioxidant medicine. Irrigation with water treated with a static magnetic field (SMF) is a possible strategy to increase rosemary yield. This technology has been applied to accelerate plant growth and increase crop quality. The aim of this study was to evaluate the content of phenolic compounds and antioxidant activity in aqueous leaf extracts from plants irrigated with SMF-treated water in comparison to control plants. Significant differences in phenolic content and antioxidant activity were observed between aqueous extracts of control plants and plants irrigated with SMF-treated water. Therefore, irrigation with SMF-treated water is a promising technology to improve the cultivation of rosemary as a raw material to obtain pharmaceutical products with high antioxidant activities.

Quantification of chlorogenic acid, rosmarinic acid, and caffeic acid contents in selected Thai medicinal plants using RP-HPLC-DAD

The chlorogenic acid, rosmarinic acid, and caffeic acid contents in 100 selected plants were determined using reversed phase high performance liquid chromatography equipped with diode array detector. The optimum condition was 0.2% phosphoric acid in water (solvent A) and methanol (solvent B) as the mobile phase, which was set at 45% B for 20 minutes at a flow rate of 1.2 mL/min. The column temperature was maintained at 30 ºC and the detection wavelength was 325 nm. Among 100 selected plants, 39.64% contained all 3 compounds, 40.54% contained 2 compounds, 14.41% contained only 1 compound, and 5.41% could not detect any of the 3 compounds. The highest contents of chlorogenic acid, rosmarinic acid, and caffeic acid were found in Lonicera japonica flowering buds, Melissa officinalis leaves, and Coffea canephora seeds at the concentration of 9.900 ± 0.004, 19.908 ± 0.171, and 1.233 ± 0.003 g/100 g of dried plant, respectively.

The neurogenic effects of rosmarinic acid in a mouse model of type 2 diabetes mellitus

There is emerging evidence for a dysregulation of insulin signaling in the brains of patients with Alzheimer's disease (AD) with overlapping molecular features to Type 2 Diabetes Mellitus (T2DM). In addition, T2DM is a known risk factor of AD. The goal of this study was to investigate the neurogenic and neuroprotective potential of rosmarinic acid (RA) in a streptozotocin (STZ)-induced combined with high fat diet (HFD) mouse model of diabetes. Animals were divided into four experimental groups (control, diabetic, diabetic + RA, RA only). Behavioral analysis was performed to assess spatial learning and anxiety levels of animals, whereas quantitative real time PCR was carried out to assess the gene expression levels of neuronal markers of neurogenesis (Ki67, DCX and NeuN). A significant decrease in memory and spatial learning was observed in the diabetic mice, which was substantially improved by RA treatment. RA also increased the gene expression of NeuN, DCX and Ki67, which were dysregulated in the diabetic model. This study proposes RA as a potential therapeutic agent to mitigate neuronal dysfunction associated with T2DM by promoting adult hippocampal neurogenesis.

Optimization of water-precipitation process in pretreatment process of Salviae Miltiorrhizae (Danshen) and Ligustrazine Hydrochloride Injection based on sequential central composite design / 中草药

Objective: To optimize the water-precipitation in the pretreatment manufacture process of Salviae Miltiorrhizae (Danshen) and Ligustrazine Hydrochloride Injection (SMLHI) based on the quality by design (QbD) concept and the sequential design. Methods: A design space was obtained by the global-type sequential design methodology combined with the fishbone-diagram risk analysis and central composite experiment based on global type sequential design method. The preliminary optimized zone of critical process parameters was extended in axial direction on the basis of the statistical result gained from the first central composite design. Then an updated multivariate linear regression model was fitted and a design space was delineated and verified. Results: According to the design space, the recommended operation space of the water-precipitation was as follows: when the pH value of the material was between 3.1 and 3.4, 3.25-5.00 times of water adding into the solution was recommened, the standing time was 7-17 h and the standing temperature was 7 ℃. Conclusion: This study introduces the sequential design into the process optimization of traditional Chinese medicine pharmaceuticals. Compared with the experimental design methods such as uniform space grid design and orthogonal design with a large number of samples, while samples cover the same feasible zone in a continuous operation form, sequential design methodology can effectively reduce the workload of the optimization experiment, avoid the waste of manpower and resources as well as guarantee the prediction ability of the fitting model.

Rosmarinic acid inhibits high glucose-induced cardiomyocyte hypertrophy by activating Parkin-mediated mitophagy / 南方医科大学学报

OBJECTIVE@#To evaluate the effect of rosmarinic acid (RA) on mitophagy and hypertrophy of cardiomyocytes exposed to high glucose (HG).@*METHODS@#Rat cardiomyocytes (H9c2) exposed to HG (25 mmol/L) were treated with 50 μmol/L RA or with both RA treatment and Parkin siRNA transfection, with the cells cultured in normal glucose (5.5 mmol/L) and HG as the controls. The expressions of PINK1, Parkin and LC3II/LC3I in the cells were detected by Western blotting. The formation of mitochondrial autophagosomes was observed by transmission electron microscope. Flow cytometry was employed to detect the level of reactive oxygen species (ROS) and apoptotic rate of the cells. The activities of respiratory chain complex enzymes were measured by spectrophotometry. Fluorescence enzyme labeling and @*RESULTS@#RA treatment significantly increased the expression levels of PINK1, Parkin and LC3-II/I (@*CONCLUSIONS@#RA can protect rat cardiomyocytes against oxidative stress injury and cardiomyocyte hypertrophy induced by HG by activating Parkin-mediated mitophagy.

Phytochemical compounds of anise hyssop (Agastache foeniculum) and antibacterial, antioxidant, and acetylcholinesterase inhibitory properties of its essential oil

Agastache foeniculum (Pursh) Kuntze (anise hyssop) is known as a medicinal and flavoring spice plant. The aim ofthis research was gas chromatography-mass spectrometry (GC-MS) analysis of anise hyssop essential oil and analysisof radical scavenging ability, antimicrobial activity, and acetylcholinesterase inhibitory activity. Phytochemicalcomposition of anise hyssop wastes and the aqueous extract obtained after steam distillation was investigated.Eight compounds were identified in essential oils by GC-MS analysis, as the major ones were phenylpropenoids[estragol—93.45% of total ion current (TIC), eugenol—0.15% of TIC, and methyl isoeugenol—2.48% of TIC]. Inthe anise hyssop wastes extract, four pentacyclic triterpenes were identified (betulin—36.1 mg/g, betulinic acid—2.4mg/g, oleanolic acid—160.0 mg/g, and ursolic acid—6.7 mg/g extract). Rosmarinic acid (50.6 mg/g extract) andflavonoids—myricetin (0.5 mg/g), luteolin (0.9 mg/g), and apigenin (0.6 mg/g) were detected by high-performanceliquid chromatography with diode-array detection analysis of aqueous extract. The anise hyssop essential oil showedstrong radical scavenging ability IC50—6.54 μl/ml. The results obtained from antimicrobial screening revealed thatessential oil possessed inhibitory activity against Staphylococcus aureus ATCC 25923, Curtobacterium flaccumfaciensPM_YT, Listeria monocytogenes, Bacillus subtilis ATCC 6633, Salmonella sp., Escherichia coli ATCC 8739, Proteusvulgaris, Pseudomonas aeruginosa ATCC 9027, Klebsiella pneumoniae, and Candida albicans, while Enterococcusfaecalis remained unaffected.