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.

牛至渣饲料发酵过程中迷迭香酸等活性成分的变化规律研究

本试验旨在研究中草药牛至渣微生物发酵过程中迷迭香酸、绿原酸、总苷、多酚、总酚酸等活性成分的变化规律,为生产制备绿色、环保的发酵饲料提供理论依据。设计单因素及正交试验考察培养基成分、菌种及发酵条件对上述5种活性成分含量的影响。单因素试验方差分析和显著性检验结果显示:总苷、总多酚和总酚酸在发酵20 d时含量最高,迷迭香酸和绿原酸在发酵15 d时含量最高。正交试验结果表明采用高含量菌种发酵中药渣,并以粗麸皮+次粉为辅料,所得饲料营养成分含量最高。其中总苷含量1.351%,迷迭香酸含量2.733%,绿原酸含量12.323%,总多酚含量21.604%,总酚酸含量0.16%。经微生物发酵后,所得牛至渣发酵饲料中总苷、迷迭香酸、绿原酸和总多酚的含量均有显著提高。该研究为生产制备以牛至渣等中药渣为原料的发酵饲料、实现牛至中药材资源的综合开发利用提供参考。

利用纳滤传质模型定量计算迷迭香酸分子状态的实验教学设计

该文设计了利用纳滤传质模型定量计算迷迭香酸分子状态的教学实验。实验以丹参中代表性的酚酸为研究对象,计算了迷迭香酸的纳滤膜通量、传质系数和截留率,构建了迷迭香酸分子态比例定量计算数学模型,对制药过程中的醇沉、树脂吸附、澄清过滤、浓缩、干燥等工艺的转移率和成分存在状态进行了相关性分析,研究了制药工艺成分量值传递的内在机制。实验结果表明,迷迭香酸存在状态对制药工艺中的成分转移率有重要影响。这一结论对提升生产过程的规范化和标准化很有意义。本实验与制药生产热点问题相结合,通过科研反哺教学,有助于增强学生的科研素养和实践能力,并实现科研应用与教学拓展的协同提升。

迷迭香酸对子痫前期大鼠炎症和内皮功能障碍的影响

目的 探究迷迭香酸(RA)对子痫前期(PE)大鼠的治疗作用及机制。方法 从孕第11天开始,对孕鼠连续4 d每天皮下注射左旋硝基精氨酸甲酯(L-NAME,200 mg/kg)建立PE大鼠模型,将30只成功建模的大鼠随机分为3组:模型组、50RA组(50 mg/kg RA)和100RA组(100 mg/kg RA),每组10只。另选10只同期孕鼠作为对照组。孕第15天时,50RA组和100RA组大鼠灌胃不同浓度RA,对照组和模型组大鼠灌胃等体积生理盐水,连续灌胃7 d。孕第21天时,检测各组大鼠的收缩压(SBP)、24 h尿蛋白、胎鼠体质量和死亡率,并对胎盘组织进行苏木精-伊红(HE)染色,对肾脏组织进行过碘酸-雪夫(PAS)染色。按照试剂盒说明检测大鼠血清超氧化物歧化酶(SOD)和丙二醛(MDA)水平。使用ELISA试剂盒检测大鼠血清中细胞因子白细胞介素(IL)-1β、IL-6、干扰素-γ(IFN-γ)和单核细胞趋化蛋白(MCP)-1的水平。通过Western blot检测胎盘组织中细胞间黏附分子-1(ICAM-1)、白细胞功能相关抗原(LFA-1)、血管细胞黏附分子-1(VCAM-1)、Toll样受体4(TLR4)和髓样分化因子88(MyD88)的蛋白表达水平。结果 与对照组相比,模型组大鼠的SBP和24 h尿蛋白水平升高(P<0.05),胎鼠体质量降低(P<0.05),胎鼠死亡率升高(P<0.05),胎盘和肾脏组织出现明显损伤,血清SOD水平降低(P<0.05),MDA水平升高(P<0.05),血清IL-1β、IL-6、IFN-γ和MCP-1水平升高(P<0.05),胎盘组织中ICAM-1、LFA-1、VCAM-1、TLR4和MyD88的蛋白表达水平升高(P<0.05)。与模型组相比,50RA组和100RA组大鼠的SBP和24 h尿蛋白水平均降低(P<0.05),胎鼠体质量均升高(P<0.05),胎鼠死亡率均降低(P<0.05),胎盘和肾脏组织形态明显改善,血清SOD水平均升高(P<0.05),MDA水平均降低(P<0.05),血清IL-1β、IL-6、IFN-γ和MCP-1水平均降低(P<0.05),胎盘组织中ICAM-1、LFA-1、VCAM-1、TLR4和MyD88的蛋白表达水平均降低(P<0.05)。与50RA组相比,100RA组大鼠的SBP和24 h尿蛋白水平降低(P<0.05),胎鼠体质量升高(P<0.05),胎盘和肾脏组织形态明显改善,血清SOD水平升高(P<0.05),MDA水平降低(P<0.05),血清IL-1β、IL-6、IFN-γ和MCP-1水平降低(P<0.05),胎盘组织中ICAM-1、LFA-1、VCAM-1、TLR4和MyD88的蛋白表达水平降低(P<0.05)。结论 RA有效减轻了PE大鼠的病理改变,RA可能通过减轻TLR4信号通路介导的炎症、氧化应激和内皮功能障碍来发挥其对PE的治疗作用。

迷迭香酸共价偶联对β-乳球蛋白结构及性质的影响

以β-乳球蛋白(β-lactoglobulin,β-LG)为对象,探究膳食多酚迷迭香酸(rosmarinic acid,RA)共价偶联对蛋白结构和性质的影响。采用自由基法和碱法制得β-LG-RA共价偶联复合物;利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、傅里叶变换红外光谱、紫外吸收光谱、荧光光谱、圆二色光谱等技术,分析RA共价偶联后β-LG结构变化规律;通过对1,1-二苯基-2-三硝基苯肼自由基和2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基清除率检测及酶联免疫吸附测试,评价β-LG与RA共价偶联前后抗氧化性和与血清特异性免疫球蛋白G(immunoglobulin G,IgG)结合能力的变化。结果表明:RA共价偶联后β-LG的二级结构由α-螺旋向β-折叠和无规卷曲转变,蛋白的二、三级结构发生改变。此外,与β-LG相比,β-LG-RA共价偶联复合物的抗氧化性显著增强,与血清特异性Ig G的结合能力显著减弱。综上,RA共价偶联通过引入酚羟基增强了蛋白的抗氧化性,改变了β-LG的结构,降低了其与血清特异性IgG的结合能力。

迷迭香酸⁃锌配合物的抗氧化和抗菌性研究

目的:研究迷迭香酸(rosmarinic acid,RA)和锌(zinc,Zn)离子络合后的结构、抗氧化性、抗菌性和生物相容性。方法:利用扫描电子显微镜、红外和紫外⁃可见光谱分析迷迭香酸锌配合物(RA⁃Zn)的结构。通过细胞增殖实验比较RA和RA⁃Zn对成骨细胞增殖活性的影响,通过铁离子还原抗氧化能力实验和2,2⁃联氮⁃二(⁃3⁃乙基苯并噻唑⁃6⁃磺酸)二铵盐[2,2′⁃azinobis(3⁃ethylbenzothiazoline⁃6⁃sulfonic acid)ammonium salt,ABTS]自由基清除实验分析两者的抗氧化性,最小抑菌浓度法比较两者对金黄色葡萄球菌的抑制活性。结果:成功合成了RA⁃Zn。作用第5天,RA⁃Zn促进成骨细胞增殖的作用优于RA(P<0.05),200μmol/L RA⁃Zn的抗氧化能力比RA更强(P<0.05)。RA浓度≥400μmol/L、而RA⁃Zn浓度≥25μmol/L时能抑制金黄色葡萄球菌活性(P<0.05)。结论:RA和Zn离子络合后抗氧化性和抗菌性增强,具有促进成骨细胞增殖的作用。

基于网络药理学与脂质组学探讨迷迭香酸缓解食物过敏的效应机制

为研究迷迭香酸缓解食物过敏的效应机制,利用网络药理学和分子对接以及脂质组学探究迷迭香酸(Rosmarinic acid,RA)缓解食物过敏的效应机制。利用数据库分析平台(TCMSP)、Swiss Target Prediction、Pharm Mapper、Super Pred和Gene Cards等获得RA的潜在效应靶点。通过软件Cytoscape和String数据库构建“活性成分-靶点”蛋白互作(PPI)网络,并采用AutoDockTools软件进行分子对接验证。进一步通过建立卵清蛋白(Ovalbumin,OVA)食物过敏小鼠模型验证RA缓解食物过敏的效应,并基于高分辨质谱对小鼠血清进行脂质代谢分析。结果表明,网络药理分析共筛选到关联靶点46个。基因本体(Geno ontology,GO)富集分析表明RA主要通过对蛋白质磷酸化、鞘糖脂、磷脂和类固醇等的结合发挥作用。京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析表明关联靶点主要富集在Th1和Th2细胞分化通路、脂代谢和炎症性肠病通路等。动物实验结果显示RA干预可以显著(P<0.05,P<0.01,P<0.001)抑制OVA致敏引起的小鼠血清过敏反应介质上升,且RA干预组小鼠血清总胆固醇(Total Cholesterol,T-CHO)和甘油三酯(Triglyceride,TG)水平下降。脂质组学分析中,在正/负离子模式下分别筛选出25/36种潜在差异脂质代谢物。对脂质差异代谢物的富集分析表明,小鼠的血清脂质代谢影响甘油磷脂代谢和鞘脂代谢通路。因此,本研究表明RA主要通过调节脂质代谢和机体炎症反应发挥缓解食物过敏的作用。

基于PI3K/Akt通路探究迷迭香酸对自身免疫性甲状腺炎大鼠的作用及机制

目的基于磷酸肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路探究迷迭香酸对自身免疫性甲状腺炎(AIT)大鼠的作用及机制。方法通过高碘水喂养联合皮下注射甲状腺球蛋白及弗氏佐剂来诱导建立AIT大鼠模型,随机分为4组:模型组、迷迭香酸低剂量(20mg/kg)组、迷迭香酸高剂量(40mg/kg)组、迷迭香酸高剂量(40mg/kg)+IGF-1(PI3K/Akt激活剂,0.75μg/kg)组,每组10只,另选10只SD大鼠,正常水喂养并皮下注射等剂量生理盐水作为对照组,以迷迭香酸和IGF-1分组处理后,ELISA测量大鼠血清甲状腺过氧化物酶抗体(TPOAb)、抗甲状腺球蛋白抗体(TGAb)水平;HE染色检测各组大鼠甲状腺组织病理损伤;免疫组织化学染色检测各组大鼠甲状腺组织CD45R表达;TUNEL染色检测各组大鼠甲状腺细胞凋亡情况;ELISA测量大鼠血清中Th细胞分泌的炎症相关因子γ干扰素(IFN-γ)、白细胞介素(IL)-4、IL-17、IL-10水平;免疫印迹实验检测各组大鼠甲状腺组织PI3K/Akt通路相关蛋白表达。结果与对照组相比,模型组大鼠TGAb、TPOAb、IFN-γ及IL-17水平、病理损伤评分、CD45R相对阳性表达、甲状腺细胞凋亡率、p-PI3K/PI3K、p-AKT/Akt明显升高(P<0.05),IL-4、IL-10水平明显降低(P<0.05);与模型组相比,迷迭香酸低剂量组、迷迭香酸高剂量组大鼠TGAb、TPOAb、IFN-γ及IL-17水平、病理损伤评分、CD45R相对阳性表达、甲状腺细胞凋亡率、p-PI3K/PI3K、p-AKT/Akt均降低(P<0.05),IL-4、IL-10水平均升高(P<0.05),且高剂量迷迭香酸作用更强;IGF-1可减弱迷迭香酸对AIT大鼠的作用。结论迷迭香酸可通过阻断PI3K/Akt通路激活来调控Th细胞因子分泌,从而抑制免疫炎症反应,减轻甲状腺组织淋巴细胞浸润及甲状腺细胞凋亡,改善AIT大鼠症状。