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OBJECTIVE To s tudy the impr ovement effects of tilianin on the atherosclerosis （AS）model mice and its potential mechanism. METHODS Eight C 57BL/6J mice were taken as the normal group. Forty ApoE－/－ mice were randomly divided into model group ，tilianin low-dose ，medium-dose and high-dose groups [ 2.1，3.5，7.0 mg/（kg·d）] and simvastatin group [positive control drug ，3.5 mg/（kg·d）]，with 8 mice in each group. Normal group was given normal diet ，and other groups were given high-lipid diet to induce AS model. At the same time ，normal group and model group were given normal saline intragastrically ， administration groups were given relevant drug intragastrically ，once a day ，for 12 consecutive weeks. The levels of TC ，TG， LDL-C，HDL-C，Ox-LDL，IL-1β，IL-6，MCP-1 and TNF-α in plasma were determined. The pathomorphological changes of the aorta in mice were observed. The positive rate of ICAM- 1，VCAM-1 and PCNA in the aorta were determined. mRNA expressions of MMP- 2，MMP-9，TGF-β1，Smad2 and Smad 3 as well as protein expressions of TGF-β1，Smad2/3 and p-Smad 2/3 were also determined in aorta of mice. RESULTS Compared with normal group ，the plasma levels of TC ，TG，LDL-C，Ox-LDL，IL-1β， IL-6，MCP-1 and TNF-α in model group were increased significantly（P＜0.01），while HDL-C level was significantly reduced （P＜0.01）. Lipid plaques were formed in the aorta ，and the plaque area was large and caused severe stenosis of the lumen. mRNA expressions of MMP- 2，MMP-9，TGF-β1，Smad2 and Smad 3 as well as positive rate of ICAM- 1，VCAM-1，PCNA and protein expression TGF-β1，Smad2/3，and p-Smad 2/3 in the aorta were significantly increased （P＜0.01）. Compared with model group ， most of above indexes of medication groups were improved significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Tilianin can inhibit the activation of TGF-β1/Smads signaling pathway and then inhibit the proliferation of vascular smooth muscle cells ，reduce ， inflammation and regulate lipid metabolism to inhibit the No.81960766） formation of AS.
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Z-VAD-FMK was combined with hypoxia-reoxygenation (H/R) injury to establish a necroptosis model of H9c2 cells to mimic the pathological changes of myocardial ischemia reperfusion injury (MIRI) in vitro and to study the effect and mechanism of tilianin against myocardial ischemia-reperfusion injury. A cell counting kit-8 (CCK-8) was used to detect cell viability, and commercial kits were used to detect lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in the cell culture supernatant. Hoechst 33342/PI immunofluorescence staining was used to detect cell death. DCFH-DA, BBcellProbeTMM61, and JC-1 probes were used to detect reactive oxygen species (ROS), mitochondrial permeability transition pore (mPTP), and mitochondrial membrane potential (MMP), respectively. An enzyme-linked immunosorbent assay (ELISA) method was used to detect the release of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6). The results show that the cell viability, SOD activity, and MMP of the model group induced by H/R injury decreased, as compared with control group, but the necroptosis rate, LDH level, and ROS release increased significantly. Furthermore, mPTP of the model group cells opened, and TNF-α, IL-1β, and IL-6 levels were significantly higher. Molecular docking modeling showed that tilianin can bind to calmodulin-dependent protein kinase II (CaMKII), and Western blot results showed that compared with control group, the expression levels of p-CaMKII and phospho-mixed lineage kinase domain-like protein increased in the model group, and tilianin could decrease the expression level of these proteins. The above results indicate that tilianin can protect H9c2 cells by inhibiting the phosphorylation of CaMKⅡ at threonine 287, protecting mitochondrial function, and inhibiting the opening of mPTP to prevent necroptosis. This study has value for research on new methods to treat H/R injury.
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Objective: To prepare sustained-release tablets of tilianin nanosuspension lyophilized powder. The factors that might influence drug release and release mechanism were studied in present study. Methods: High pressure homogenization method was used to prepare tilianin nanosuspension. Lactose and mannitol (3:1) were employed as freeze-drying protective agent to prepare lyophilized powder. HPMC was used as framework material to prepare sustained-release tablets of tilianin nanosuspension lyophilized powder. Based on single factor test, the effects of proportion and amounts of HPMC K4M and HPMC K15, amounts of PEG 4000 and magnesium stearate on in vitro drug release of sustained-release tablets were investigated. Orthogonal test was designed to gain the optimum prescription. Results: The particle size and zeta potential of tilianin nanosuspension were (164.41 ± 9.72) nm and (-37.21 ± 2.38) mV, respectively. The particle size and zeta potential of re-dispersed freeze-drying products were (211.83 ± 11.26) nm and (-31.66 ± 2.92) mV, respectively. The optimum prescription was as follow: the proportion and amounts of HPMC K4M and HPMC K15 were 2:1 and 40 mg, amounts of PEG 4000 was 20 mg, and amounts of magnesium stearate were 0.5%. Sustained release tablets of tilianin nanosuspension were well accorded with Higuchi kinetics model. The equation was Mt/M∞ = 0.286 8 t1/2-0.073 8, r2 = 0.981 4. And the cumulative release could achieve 92.36% in 12 h. The drug release from the tablets was controlled by diffusion and degradation of the matrix. Conclusion: The preparation technology of sustained release tablets of tilianin nanosuspension lyophilized powder has good reproducibility. This sustained release tablets could control the release of tilianin
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Objective To optimize the preparation technology of transcription activator (TAT) and polyethylene glycols (PEG) co-modified tilianin-loaded composite phospholipid liposome (TAT & PEG tilianin CPL, T&PTCPL) and investigate its protective effect on cardiomyocytes. Methods The composite phospholipid liposome was prepared by thin film-ultrasonic method. A three- factor, three-level Box-Behnken experimental design was employed. The weight ratio of total phospholipid to tilianin (X1), the concentration of DSPE-PEG2000-TAT (X2), and hydration volume (X3) were observed. The encapsulation efficiency (Y1), particle size (Y2), and polydispersion coefficient (Y3) were evaluated to optimize optimal formula. In addition, hypoxia/reoxygenation model was established with Na2S2O4 in H9C2 cells. Superoxide dismutase (SOD) activity, malonaldehyde (MDA) level and release of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) were assessed to evaluate the effect of T&PTCPL, meanwhile, the in vitro release rate (dynamic dialysis method) and absorption rate of tilianin and T&PTCPL in Caco-2 cell were examined. Results The optimal formula was as following: X1 = 20, X2 = 1.7%, and X3 = 3.2 mL; The encapsulation efficiency was (86.62 ± 2.51)%, particle size was (149.7 ± 8.2) nm and PDI was 0.15 ± 0.05. Compared with model group, T&PTCPL and tilianin groups increased SOD activity, inhibited level of MDA, LDH and CK-MB leakage (P < 0.05), and the effect of T&PTCPL group was better than tilianin group, meanwhile, T&PTCPL was completely released at 48 h, with a cumulative release of 88.65%, and Caco-2 cells had better absorption of T&PTCPL. Conclusion The Box-Behnken design is suitable for optimizing the formulation of T&PTCPL, and the observed responses are in close agreement with the predicted values of the mathematic models; Moreover, T&PTCPL shows a better sustained release effect in vitro release, which promots the absorption of tilianin in Caco-2 cells and suggests that T&PTCPL may have protective effect on myocardial ischemia reperfusion injury.
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OBJECTIVE:To study the effects of tilianin(TIL)on brain tissue in rats with cerebral ischemia-reperfusion injury. METHODS:Totally 120 male SD rats were randomly divided into sham operation group(0.9% sodium chloride solution),model group(0.9% sodium chloride solution),nimodipine group(32 mg/kg)and TIL low-dose and medium-dose,high-dose groups(4, 8,16 mg/kg),with 20 rats in each group. The rats were given relevant medicine intragastrically,once a day,for consecutive 7 d. 15 min after last medication,cerebral ischemia-reperfusion injury model was established by reforming suture-occluded method. The neurological deficit score in rats were evaluated, and percentage of cerebral infarction volume of rats was determined. Histopathological changes of brain tissue were observed by HE staining. The activities of SOD,CAT and LDH,MDA content in cerebral tissue of rats were determined. The expression of calcitonin gene-related peptide(CGRP)and peripheral vascular endothelial growth factor receptor 2 (VEGFR2) protein were determined by Western blot assay. RESULTS:Compared with sham operation group,neurological deficit score and percentage of cerebral infarction volume of model group were increased significantly(P＜0.01);the nerve cells in brain tissue were significantly reduced and the interstitial edema was obvious. SOD and CAT activities were decreased significantly,LDH activity was increased significantly,MDA content was decreased significantly,protein expression of CGRP and VEGFR2were increased significantly(P＜0.05 or P＜0.01). Compared with model group,neurological deficit score of nimodipine group,TIL medium-dose and high-dose groups were decreased significantly;percentage of cerebral infarction volume was decreased significantly (P＜0.05 or P＜0.01);above pathological conditions of cerebral tissue in rats were relieved significantly;SOD and CAT activities were strengthened significantly,MDA content and LDH activities were decreased significantly,protein expression of CGRP and VEGFR2were increased significantly (P＜0.05 or P＜0.01). CONCLUSIONS: TIL has certain protective effects on cerebral ischemia-reperfusion injury model rats,and its mechanism may be related to the up-regulation of CGRP and VEGFR2expression.
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OBJECTIVE: To prepare nano-micelles with amphiphilic self-assembly poly (ethylene glycol)-co-poly (propylene sulfide) (PEG-PPS) copolymer as carrier to study the release characteristics of tilianin and investigate its activity to against H9c2 cell apoptosis in vitro. METHODS: An amphiphilic diblock PEG-PPS polymer was used as a carrier material to prepare the tilianin-containing nano-micelles by solvent evaporation. The morphology, particle size and distribution, drug loading and encapsulation rate and in vitro drug release behavior were characterized, H9c2 rat myocardial cell injury model was established by hypoxia/reoxygenation process. Using propranolol (Pro) as a positive control, the morphology of injured cardiomyocytes was observed by microscope. Cell proliferation and cell apoptosis was detected to evaluate the protective effect of blank micelles, tilianin and tilianin loaded nano-micelles on H9c2 cells induced by hypoxia/reoxygenation. RESULTS: Tilianin-loaded nano-micelles was spherical with uniform particle size distribution. The drug loading was 3.82%. The average particle diameter of tilianin-loaded nano-micelles was 137 nm, polydispersity coefficient was 0.162 and the encapsulation efficiency was 91.45%. In vitro drug release studies showed that there was no drug-induced burst release of tilianin-containing nano-micelles and sustained release characteristics, and the presence of hydrogen peroxide significantly promoted the release of tilianin from the nano-micelles. In vitro cytotoxicity experiments showed that when the concentration of tilianin 5 μg•mL-1, the cell viability of tilianin-loaded nano-micelles was significantly higher than the corresponding concentration of tilianin and PEG-PPS polymer nano-micelles. In vitro anti-apoptotic activity experiments show that tilianin-loaded nano-micelles on H9c2 cell apoptosis induced by hypoxia-reoxygenation have a significant inhibitory effect and was provided inhibition of apoptosis with propranolol. CONCLUSION: Tilianin-loaded nano-micelles have uniform particle size and distribution, sustained release and oxidation characteristics, has a significant protective and apoptosis-inhibiting effect on H9c2 cell injury induced by hypoxia-reoxygenation, which can be used as a promising drug delivery system for the treatment of myocardial ischemia-reperfusion injury.
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Objective: To prepare and optimize tilianin loaded solid lipid nanoparticles (T-SLNs), and investigate the physicochemical properties, absorption and transport behavour of T-SLNs in vitro. Methods: T-SLNs were prepared by high shear homogenization followed by ultrasonication and optimized by central composite design and response surface methodology. In the study, the physicochemical properties of T-SLNs including size, polydispersity (PDI), Zeta potential, shape, entrapment efficiency and release profile in vitro were investigated, the absorption and transport behavour of T-SLNs in Caco-2 cell model were also measured. Results: The optimum formulation of T-SLNs consisted of: drug/lipid of 0.11, soy lecithin/lipid of 1.26, and content of tween-80 was 5.05%. The prepared T-SLNs were spherical and uniform with the mean particle diameter at (86.40 ± 0.62) nm, PDI (0.165 ± 0.080) and Zeta potential of (-24.2 ± 0.6) mV, respectively. The average EE was (89.81 ± 1.07)%, and the release in vitro showed that tilianin was released about (98.72 ± 1.57)% in 48 h. Besides, the absorption and transport assays of T-SLNs in Caco-2 cells model indicated that T-SLNs had a higher absorption and transport than tilianin. Conclusion: The method of high shear homogenization followed by ultrasonication is suitable for T-SLNs preparation. The optimal T-SLNs have a smaller particle size and high EE. Moreover, in the same concentration of tilianin, the absorption and transport amounts of T-SLNs in Caco-2 cell model were higher than tilianin.
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Objective: To study the chemical constituents from plant of Artemisia frigida Willd. and its pharmacological activities. Methods: The compounds were isolated and purified by various chromatography methods. All compounds were identified by spectroscopic and chromatographic techniques, and were screening for their PPARγ activating activity and PTP1B inhibitory activity. Results: There were 26 compounds isolated from the plant of A. frigida and identified as pectolinarigenin (1), jaceosidin (2), chrysoeriol (3), tricin (4), 3-oxogermacra-1(10),11(13)-dien-6α,12-olide (5), achillin (6), 1,10β-epoxyachillin (7), scoparone (8), 4-hydroxyacetophenone (9), chrysosplentin (10), jaceidin (11), 11α,13-dihydroyomogin (12), chrysanthemin A (13), eupatilin (14), eupatrin (15), artemorin (16), 6-methoxytricin (17), hanphyllin (18), cirsimaritin (19), ridentin (20), desacetylmatricarin (21), subchrysine (22), luteolin (23), caffeic acid (24), agastachoside (25), and tilianin (26). Conclusion: Compounds 5, 7, 11, 18, 25, and 26 are firstly isolated from the Artemisia. Compounds 10, 12, 13, 16, 17, and 22 are firstly isolated from this plant. Compounds 2 and 15 exhibited weak activity of PPARγ. Compounds 1 and 3 had inhibitory effect on PTP1B.
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Objective: To investigate the technology for the separation and purification of extract in Dracocephalum moldevica (EDM) by macroporous resin. Methods: Static and dynamic adsorption-desorption were used to select the best one from seven different type macroporous resins; With the content of total flavonoids, tilianin, luteolin-7-O-β-D-glucuronide, and rosmarinic acid as indexes, the purification technology parameters of EDM were optimized. Results: HPD600 resin showed the best purifying profile, its optimum technology conditions were as follows: The optimum concentration of the sample liquid was 0.08 g/mL equivalent to raw material, the resin column diameter-height ratio was 1:9, the amount of used adsorption was 0.32 g dried medicinal herb/mL resin, sample flow rate was 1.5 BV/h, and adsorption time was 12 h. In the course of elution, the resin column chromatography was eluted with 6 BV of 70% ethanol after removing impurities with 4 BV of water by flow rate of 1.5 BV/h. The contents of total flavonoids, tilianin, luteolin-7-O-β-D-glucuronide, and rosmarinic acid were more than 53%, 5.5%, 4.7%, and 2.5%. Conclusion: Macroporous resin HPD600 is suitable to separate and purify EDM.
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Objective: Taking luteolin-7-O-β-D-glucuronide, rosmarinic acid, and tilianin as indexes, the concentration of extracts from Dracocephalum moldavica (EDM) and their index components in various media, surfactant solution, and apparent oil/water partition coefficient (P) was detected in different pH values, then the stability of EDM in artificial gastrointestinal fluid was investigated. This research would give the reference for the selection and preparation of further novel formulation. Methods: Apparent solubility of EDM in various media and surfactant solution was determined by precipitation method and a shake flask method was used to determine the P of octanol-water/phosphate buffer salt, the stability of EDM in artificial gastrointestinal fluid was investigated. HPLC was adopted to determine the concentration of the index components with Shim-pack ODS column (250 mm×4.6 mm, 5 μm), mobile phase of acetonitrile (A)-0.5% formic acid aqueous solution (B) for gradient elution (0-30 min, 15% A; 30-55 min, 15%→25% A; 55-80 min, 25%→35% A) as the mobile phase at a flow rate of 1.0 mL/min. The detection wavelength was set at 324 nm and the column temperature was maintained at 35℃. Results: At 37℃, the equilibrium solubility of the three index components in acid buffer solution increased obviously in water, but in alkaline the buffer solution of the equilibrium solubility decreased with pH increasing. In 32 g/L sodium dodecyl sulfate solution, the equilibrium solubility of luteolin, rosmarinic acid, and tilianin increased to 1679.61, 1249.2, and 2765.27 μg/mL. The P of luteolin-7-O-β-D-glucuronide, rosmarinic acid, and tilianin were 0.1731 (lgP=-0.7618), 0.068 4 (lgP=-1.1650), and 1.0829 (lgP=0.0346), and increased as pH rising. EDM in artificial gastrointestinal fluid was stable (except luteolin). Conclusion: The methods can be used to determine the apparent solubility, the P value of the extracts and their index components; The EDM in artificial gastrointestinal fluid is stable. The research would give the reference for the selection and preparation of the further novel formulation.
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OBJECTIVE: To develop an RP-HPLC method for simultaneous determination of rosmarinic acid, tilianin, luteolin-7-O-β-D-glucuronide, apigenin-7-O-β-D-glucuronide, and diosmetin-7-O-β-D-glucuronide in different parts ofDracocephalum moldevica L. METHODS: The five constituents were measured on a Shim-pack ODS column(4.6 mm×250 mm, 5 μm)with gradient elution of acetonitrile (A)-0.5% formic acid aqeous solution (B) (0-30 min, 17%A;30-60 min, 17%-28%A; 60-70 min, 28%A) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 330 nm, and the column temperature was maitained at 35℃. RESULTS: The linear ranges of rosmarinic acid, tilianin, luteolin-7-O-β-D-glucuronide, apigenin-7-O-β-D-glucuronide, and diosmetin-7-O-β-D-glucuronide were 4.2-126 μg·mL-1 (r=0.999 2), 7.84-235.2 μg·mL-1 (r=0.999 3), 3.048-91.44 μg·mL-1(r=0.999 4), 1.472-44.16 μg·mL-1(r=0.999 4), and 2.816-84.48 μg·mL-1 (r=0.999 2), respectively. The average recoveries (RSD) of the five compounds were 98.97%(1.03%), 99.90%(0.92%), 99.89% (1.75%), 99.55% (0.98%), and 99.76%(1.19%) (n=6), respectively. CONCLUSION: The developed method is accurate and precise, which can be used for the quality control of different parts of Dracocephalum moldevica L.The RESULTS: of content determination indicate that the five compounds exist in all the parts of Dracocephalum moldevica L., but the mass fractions are obviously different.
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To study the extraction technology for the active constituents in Dracocephali Moldavici Herba (the aerial parts of Dracocephalum moldevica) and to compare their contents Dracocephali Moldavici Herba from various habitats. The contents of luteolin-7-O-glucuronide (I), apigenin-7-O-glucuronide (II), rosmarinic acid, diosmetin-7-O-glucuronide (III), tilianin, and acacetin-7-O-glucuronide (IV) in Dracocephali Moldavici Herba were measured using HPLC method. Based on single-factor test, and the influence of the extracting menstrua, menstruum dosage, and extracting time were investigated using orthogonal design method. Based on the optimal technology, the contents of I, II, rosmarinic acid, III, tilianin, and IV in Dracocephali Moldavici Herba from various habitats were compared. The optimal conditions for the extracting for one time, each time for 5 h with 50 times of amount of 40% ethanol. D. moldevica produced in Jimusar, Xinjiang had the higher contents of I, II, rosmarinic acid, III, tilianin, and acacetin-7-O-glucuronide. This extraction technology is reasonable, stable, and feasible. The contents of I, II, rosmarinic acid, III, tilianin, and IV in Dracocephali Moldavici Herba from different habitats have some differences. The multi-index contents of I, apigenin-7-O-glucuronide, rosmarinic acid, III, tilianin, and IV could reflect the quality of Dracocephali Moldavici Herba more comprehensively.
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OBJECTIVE@#To determine the depressant-like effects and the possible mechanism of action of tilianin isolated from active methanol extract of Agastache mexicana (A. mexicana). Also, to establish the pharmacophoric requirements of tilianin, as a possible ligand of GABAA/BZD receptor, by the alignment of diazepam, CGS-9896 and diindole, using a previously described pharmacophoric model.@*METHODS@#Tilianin (30 to 300 mg/kg, ip. and 300 mg/kg, po.) and methanol crude extract (10 to 300 mg/kg, ip. and 300 mg/kg po.) from A. mexicana were evaluated for potential sedative and anxiolytic-like response drugs by using open-field, hole-board, cylinder of exploration, plus-maze and sodium pentobarbital-induced hypnosis mice methods.@*RESULTS@#Methanol extract and tilianin showed anxiolytic-like activity from a dosage of 30 mg/kg, ip. or 300 mg/kg, po. and were less potent than diazepam 0.1 mg/kg, a reference anxiolytic drug used. Moreover, depressant activity of both potentiates sodium pentobarbital (SP)-induced sleeping time. The anxiolytic-like effect of 30 mg/kg ip. observed for the extract and tilianin, by using the plus-maze model, was partially prevented in the presence of flumazenil (a GABAA/BZD antagonist, 5 mg/kg ip.) but not in the presence of WAY 100635 (a selective 5-HT1A receptor antagonist, 0.32 mg/kg, ip.). Pharmacophoric modeling alignments of three agonist of GABAA/BZD allow identify seven chemical features. Tilianin contains six of the seven features previously determined.@*CONCLUSIONS@#Results indicate that tilianin is one of the bioactive metabolites in the anxiolytic-like activity of A. mexicana, reinforcing its central nervous system uses, where GABAA/BZD, but not 5-HT1A, receptors are partially involved.
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Objective:To determine the depressant-like effects and the possible mechanism of action of tilianin isolated from active methanol extract ofAgastache mexicana(A. mexicana).Also, to establish the pharmacophoric requirements of tilianin, as a possible ligand of GABAA/BZD receptor, by the alignment of diazepam, CGS-9896 and diindole, using a previously described pharmacophoric model.Methods:Tilianin (30 to 300 mg/kg,ip.and 300 mg/kg, po.)and methanol crude extract (10 to 300 mg/kg,ip. and 300 mg/kgpo.)fromA. mexicana were evaluated for potential sedative and anxiolytic-like response drugs by using open-field, hole-board, cylinder of exploration, plus-maze and sodium pentobarbital-induced hypnosis mice methods.Results:Methanol extract and tilianin showed anxiolytic-like activity from a dosage of 30 mg/kg,ip.or 300 mg/kg,po.and were less potent than diazepam 0.1 mg/kg, a reference anxiolytic drug used. Moreover, depressant activity of both potentiates sodium pentobarbital (SP)-induced sleeping time. The anxiolytic-like effect of 30 mg/kgip.observed for the extract and tilianin, by using the plus-maze model, was partially prevented in the presence of flumazenil (a GABAA/BZD antagonist, 5 mg/kgip.)but not in the presence of WAY 100635 (a selective 5-HT1A receptor antagonist, 0.32 mg/kg,ip.).Pharmacophoric modeling alignments of three agonist of GABAA/BZD allow identify seven chemical features. Tilianin contains six of the seven features previously determined.Conclusions:Results indicate that tilianin is one of the bioactive metabolites in the anxiolytic-like activity ofA. mexicana, reinforcing its central nervous system uses, where GABAA/BZD, but not 5-HT1A, receptors are partially involved.
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Objective: To determine the depressant-like effects and the possible mechanism of action of tilianin isolated from active methanol extract of Agastache mexicana (A. mexicana). Also, to establish the pharmacophoric requirements of tilianin, as a possible ligand of GABAA/BZD receptor, by the alignment of diazepam, CGS-9896 and diindole, using a previously described pharmacophoric model. Methods: Tilianin (30 to 300 mg/kg, ip. and 300 mg/kg, po.) and methanol crude extract (10 to 300 mg/kg, ip. and 300 mg/kg po.) from A. mexicana were evaluated for potential sedative and anxiolytic-like response drugs by using open-field, hole-board, cylinder of exploration, plus-maze and sodium pentobarbital-induced hypnosis mice methods. Results: Methanol extract and tilianin showed anxiolytic-like activity from a dosage of 30 mg/kg, ip. or 300 mg/kg, po. and were less potent than diazepam 0.1 mg/kg, a reference anxiolytic drug used. Moreover, depressant activity of both potentiates sodium pentobarbital (SP)-induced sleeping time. The anxiolytic-like effect of 30 mg/kg ip. observed for the extract and tilianin, by using the plus-maze model, was partially prevented in the presence of flumazenil (a GABAA/BZD antagonist, 5 mg/kg ip.) but not in the presence of WAY 100635 (a selective 5-HT1A receptor antagonist, 0.32 mg/kg, ip.). Pharmacophoric modeling alignments of three agonist of GABAA/BZD allow identify seven chemical features. Tilianin contains six of the seven features previously determined. Conclusions: Results indicate that tilianin is one of the bioactive metabolites in the anxiolytic-like activity of A. mexicana, reinforcing its central nervous system uses, where GABAA/BZD, but not 5-HT1A, receptors are partially involved.
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OBJECTIVE: To establish an HPLC method for simultaneous detemination the contents of rosmarinic acid (phenylpropanoids), luteolin-7-O-β-D-glucuronide and tilianin (flavonoids) in Dracocephalum moldavica L. Extract. METHODS: The samples were separated on Purospher® STARLP RP-18 endcapped column(4.6 mm × 250 mm, 5 μm) by gradient elution with acetonitrile (A)-0.5% formic acid solution(B) (0-28 min, 20% A; 28-55 min, 20%→28% A; 55-80 min, 28%→30% A)as the mobile phase at a flow ratio of 1.0 mL · min-1. The detection wavelength was set at 330 nm and the column temperature was maintained at 35℃. RESULTS: The calibration curves of rosmarinic acid, luteolin-7-O-β-D-glucuronide, tilianin were in good linearity over the ranges of 2.184-21.84 μg · mL-1 (r =0.999 6), 3.14-31.84 μg · mL-1(r =0.999 6), 7.9-39.5 μg · mL-1(r =0.999 5) respectively. The average recoveries were 99.62%, 99.64% and 100.12% with RSD values of 1.27%, 1.05% and 1.09%. CONCLUSION: The method is reliable, simple, and accurate, and can be used for the comprehensive quality control of Dracocephalum moldavica L. extract.
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Objective: To study the chemical constituents from Mentha haplocalyx. Methods: Compounds were isolated and purified by various chromatographic techniques, and their structures were identified by spectral analysis. Results: Eleven compounds were isolated and identified, including menthalignin (1), 5-hydroxy-6, 7, 8, 4'-tetramethoxyflavone (2), betulinic acid (3), hesperetin 7-O-β-D-glucopyranoside (4), gentisic acid 5-O-β-D-(6'-salicylyl)-glucopyranoside (5), linarin (6), acatin (7), 5-hydroxy-6, 7, 8, 3', 4'-pentamethoxyflavone (8), 5, 6, 4'-trihydroxy-7, 8-dimethoxyflavone (9), tilianin (10), and agastachoside (11). Conclusion: Compound 1 is a new lignan named menthalignan; compound 2 is a new natural product; compound 4 is isolated from the plants in Labiatae for the first time, and compounds 3 and 5 are firstly reported in the plants of Mentha L.
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We investigated the effect of tilianin upon inducible nitric oxide synthesis in the plasma of low-density lipoprotein receptor knock-out (Ldlr-/-) mice fed with high cholesterol diet and in primary peritoneal macrophages of Ldlr-/- mice. High cholesterol diet induced nitric oxide production in the plasma of Ldlr-/- mice. Tilianin reduced the level of nitric oxide (NO) in plasma from Ldlr-/- mice induced by the high cholesterol diet. Tilianin also inhibited the NO production from the primary culture of peritoneal macrophages treated with lipopolysaccharide. The inhibition of NO production was caused by the suppression of inducible nitric oxide synthase (iNOS) gene expression in peritoneal macrophages isolated from Ldlr-/- mice. Moreover, tilianin inhibited the transcriptional activation of iNOS promoter that has NF-kappa B binding element. Thus, these results provide the first evidence that tilianin inhibit iNOS expression and production of NO and may act as a potential anti-inflammatory agent.