The Protective Role of Magnoliae Flos in Preventing Ovotoxicity and Managing Ovarian Function: An In Vitro and In Vivo Study.

Magnoliae Flos (MF) is a medicinal herb widely employed in traditional medicine for relieving sinusitis, allergic rhinitis, headaches, and toothaches. Here, we investigated the potential preventive effects of MF extract (MFE) against 4-vinylcyclohexene diepoxide (VCD)-induced ovotoxicity in ovarian cells and a mouse model of premature ovarian insufficiency (POI). The cytoprotective effects of MFE were assessed using CHO-K1 or COV434 cells. In vivo, B6C3F1 female mice were intraperitoneally injected with VCD for two weeks to induce POI, while MFE was orally administered for four weeks, beginning one week before VCD administration. VCD led to a significant decline in the viabilities of CHO-K1 and COV434 cells and triggered excessive reactive oxygen species (ROS) production and apoptosis specifically in CHO-K1 cells. However, pretreatment with MFE effectively prevented VCD-induced cell death and ROS generation, while also activating the Akt signaling pathway. In vivo, MFE increased relative ovary weights, follicle numbers, and serum estradiol and anti-Müllerian hormone levels versus controls under conditions of ovary failure. Collectively, our results demonstrate that MFE has a preventive effect on VCD-induced ovotoxicity through Akt activation. These results suggest that MFE may have the potential to prevent and manage conditions such as POI and diminished ovarian reserve.

Herbal Textual Research on Magnoliae Flos in Famous Classical Formulas / 中国实验方剂学杂志

By reviewing ancient materia medica and combining with modern literature， the textual research of Magnoliae Flos has been conducted to verify the name， origin， producing area， harvesting and processing methods， in order to provide basis for the selection and use of this herb in the development of famous classical formulas. After the textual research， it could be seen that the correct name of Magnoliae Flos was Xinyi in the past dynasties， meaning spicy flower buds. The main original plants used in past dynasties are Magnolia denudata and M. biondii. The history of the research on its scientific name in recent times is complicated， many foreign scholars have given several different scientific names， but most of them are inconsistent with the actual situation of Magnoliae Flos used in ancient China， because foreign scholars failed to collect the original plants of Magnoliae Flos for accurate identification. Before the Ming dynasty， Magnoliae Flos was mainly produced in Shaanxi， and then the recorded production areas gradually increased. After the founding of the People's Republic of China， the products produced in Henan named M. biondii were highly respected， and Henan was regarded as authentic producing area， and because of the collection and distribution through Yuzhou， it was customarily called Huichunhua. In ancient times， the harvesting period of Magnoliae Flos mostly concentrated in the first and second months of the lunar calendar， and the flower buds of M. biondii were also recommended to be used as medicine， but nowadays the flower buds are mostly collected in winter and spring， and those with dry buds， large size， yellow-green color， tight inner petals， fragrant smell， and no impurities are preferred. In the past dynasties， raw products were the mainstream， and there were also frying， soaking and so on. Based on the results， it is suggested that the flower buds of M. biondii should be used in the development of famous classical formulas. If the original formula does not specify the processing requirements， the raw products can be used as medicine.

Study on the extraction process of Xinyi Powder by ambi-extracting and the inclusion process of volatile oil / 国际中医中药杂志

Objective:To screen the optimal process of ambi-extracting of Xinyi Powder and inclusion of volatile oil.Methods:Single factor experiment was used to optimize the extraction process of Xinyi Powder by taking crushing particle size, extraction times, the amount of water added and extraction time as the investigation factors. L 9(3 4) orthogonal test was used to optimize the inclusion process of volatile oil in Xinyi Powder. Results:The optimal extraction process of ambi-extracting of Xinyi Powder was as follows: the slices were not crushed, 10 times the amount of water was added, and extracted for 3 hours; the best inclusion process of volatile oil as follows: β-cyclodextrin:water=1:25, β-cyclodextrin:volatile oil=6:1, inclusion temperature 35 ℃, inclusion time 3 hours.Conclusion:The ambi-extracting process and volatile oil inclusion process are simple, stable and feasible.

Anti-Inflammatory and Antioxidant in Vitro Activities of Magnoliae Flos Ethanol Extract.

This study evaluated Magnoliea Flos ethanol extract (MFE) as a potential natural anti-inflammatory and antioxidant in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and in vitro antioxidant assays. MFE (10, 30, and 50 µg/mL) dose-dependently inhibited LSP-induced nitric oxide production, which is mediated by down-regulating gene and protein expression of inducible nitric oxide synthase and cyclooxygenase-2. MFE also down-regulated both gene and protein expression of nuclear factor-kappa B and its downstream genes, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), compared with vehicle-treated cells. As a result, MFE treatment of LPS-stimulated macrophages significantly suppressed release of pro-inflammatory cytokines, such as TNF-α and IL-6. The antioxidant in vitro test revealed 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activities of MFE (0.25∼5 mg/mL) of 16.62% to 75.17% and 38.54% to 92.91%, respectively. The ferric reducing antioxidant ability of MFE was 0.54 mM to 2.14 mM. Overall, MFE exhibited antioxidant activity and an effective anti-inflammatory response in LPS-stimulated macrophages, which is potentially valuable for application as a natural functional material.

Magnoliae Flos Essential Oil as an Immunosuppressant in Dendritic Cell Activation and Contact Hypersensitivity Responses.

Magnoliae Flos is a commonly used traditional medicinal material in Asia. It is used to treat sinusitis, nasal congestion, and hypersensitive skin. Because Magonlia Flos was described as an aromatic material in ancient Chinese texts, we hypothesized that its essential oil may be used to treat immune disorders. Dendritic cells (DCs), regarded as a major target of immunomodulators to control immune responses, play a critical role in the adaptive immune response. In this study, Magnoliae Flos essential oil (MFEO) decreased the production of the cytokines TNF-α, IL-6, and IL-12p70 in lipopolysaccharide (LPS)-stimulated DCs. It also suppressed the surface markers MHC II, CD80, and CD86 in LPS-stimulated DCs. Animal models demonstrated that the 2,4-Dinitro-1-fluorobenzene (DNFB) inducing a contact hypersensitivity response was inhibited following treatment with MFEO. In addition, MFEO inhibited the infiltration of T cells in the ears of DNFB-induced mice. To explore its bioactive compounds, the components of MFEO were analyzed using gas chromatography (GC) and GC-mass spectrometry. The results revealed that the major compounds in MFEO are camphor and 1,8-cineole. Additional DC bioassays confirmed that these compounds substantially suppressed cytokine production in LPS-induced DCs. Therefore, we demonstrated that MFEO exhibits an immunosuppressive effect both in vivo and in vitro, and camphor and 1,8-cineole may be the major components responsible for its immunosuppressive ability. The findings indicate that MFEO has the potential to be developed as a new immunosuppressant for excessive diseases.

Comparative study of volatile components of Magnoliae Flos, Magnoliae Flos kernels, and Magnoliae Flos bracts / 中草药

Objective To analyze the volatile components of Magnoliae Flos, Magnoliae Flos kernels, and Magnoliae Flos bracts and compare the differences of these components in order to provide a theoretical basis for the usage of kernels of Magnolia Flos in classical famous prescriptions. Methods The volatile components were extracted by steam distillation and analyzed by gas chromatography-mass spectrometry (GC-MS). The volatile components in different parts of Magnoliae Flos were identified by searching the retention index in the mass spectrometry database. The relative content of volatile components was calculated by peak area normalization method, and the data of GC-MS was further proceeded by principal component analysis (PCA). Results The average volatile components content in Magnoliae Flos, Magnoliae Flos kernels, and Magnoliae Flos bracts was 1.70%, 6.79%, and 0.45%, respectively. The numbers of compounds identified by GC-MS in those three different parts were 87, 82, and 85, respectively. These compounds mainly included terpene olefins and alcohols type chemicals. PCA method was then applied to process the GC-MS data and distinguish different parts and habitats of Magnoliae Flos. Conclusion The volatile components in the kernels of Magnoliae Flos were generally the same as those of Magnoliea Flos, while the content of which was much higher than those of whole flower bud of Magnoliae Flos. The results indicated that it was rational to use the kernels of Magnolia Flos in classical famous prescriptions.

Epimagnolin A, a tetrahydrofurofuranoid lignan from Magnolia fargesii, reverses ABCB1-mediated drug resistance.

BACKGROUND: Epimagnolin A is an ingredient of the Chinese crude drug Shin-i, derived from the dried flower buds of Magnolia fargesii and Magnolia flos, which has been traditionally used for the treatment of allergic rhinitis and nasal congestion, empyema, and sinusitis. The pharmacokinetic activity of epimagnolin A remains to be evaluated. PURPOSE: In this study, we examined the possible interactions of epimagnolin A with human ATP-binding cassette (ABC) transporter ABCB1, a membrane protein vital in regulating the pharmacokinetics of drugs and xenobiotics. STUDY DESIGN/METHODS: The interaction of epimagnolin A with ABCB1 was evaluated in calcein, ATPase, and MTT assays by using Flp-In-293/ABCB1 cells and purified ABCB1 and simulated in molecular docking studies. RESULTS: Epimagnolin A inhibited calcein export by Flp-In-293/ABCB1 cells in a concentration-dependent manner in a calcein assay. ATPase assay revealed a concentration-dependent stimulation of the ATPase activity of ABCB1 by epimagnolin A. Epimagnolin A also showed saturation kinetics in the relationship between the compound-stimulated ATPase activity and the compound concentration, suggesting Michaelis-Menten kinetics similar to those of the control drug, verapamil. Km and Vmax values were calculated from Hanes-Woolf plots of (compound concentration) × (compound-stimulated ATPase activity)-1 vs. (compound concentration); the Km of epimagnolin and verapamil was 42.9 ±â€¯7.53  µM and 12.3 ±â€¯4.79  µM, respectively, and the corresponding Vmax values were 156 ±â€¯15.0  µM and 109 ±â€¯3.18  µM. Molecular docking studies on human ABCB1 showed that epimagnolin A docked to the same binding pocket as verapamil, and 3-(4,5-dimethyl-2-thiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays showed that the sensitivities of Flp-In-293/ABCB1 cells against anti-cancer drugs were enhanced upon exposure to 10  µM epimagnolin A. CONCLUSION: These results strongly suggest that epimagnolin A affects the transport activity of ABCB1 as a substrate.

Study on GC fingerprint of Magnoliae Flos and Asari Radix et Rhizoma in Liujing Toutong Tablets / 中草药

Objective To establish the fingerprint of Magnoliae Flos and Asari Radix et Rhizoma of Liujing Toutong Tablets (LTT), provide the basis for the evaluation of the quality of LTT. Methods Aglient DB-WAX (20 m × 0.18 mm, 180 μm) gas chromatography column, select the appropriate temperature conditions, the determination of the 11 batch of Magnoliae Flos, Asari Radix et Rhizoma, the use of SPSS clustering analysis and chromatographic fingerprint evaluation system (2004A version) the establishment of Xinyi medicine fingerprints, and through the Agilent 7890A-5975C gas quality for instrument on the common peaks were identified. Results The 11 batches of samples were measured and SPSS cluster analysis of 10 batches of medicinal materials were clustered into one group, and the establishment of Xinyi medicine GC fingerprint, the similarity was more than 0.9, a total of 22 common peaks, which belongs to the whole party into 14 components, medicinal unique ingredients 8; 10 batches of Asarum medicinal materials were clustered into one group, and the establishment of Asarum GC fingerprint, the similarity was more than 0.9, a total of 11 common peaks, which belongs to the whole Party composition for 4, 7 for a particular component of medicinal materials. Conclusion This method has strong specificity, high accuracy, good repeatability, and provide the basis for quality control of Herba Magnoliae Flos and Asari Radix et Rhizoma.

Effect of HP-β-CD on Solubility of 1,8-Oxido-p-menthane in Volatile Oil of Magnoliae Flos and Study on the Extraction Technology of Xinzhi Nasal Drops / 中国药房

OBJECTIVE:To study the effect of HP-β-CD on solubility of 1,8-oxido-p-menthane in volatile oil of magnoliae flos,and optimize the extraction technology of Xinzhi nasal drops. METHODS:Steam distillation method was adopted to extract the volatile oil in magnoliae flos. Using the content of 1,8-oxido-p-menthane in volatile oil of magnoliae flos as index,HPLC was adopted to determine the effects of HP-β-CD with different concentration on solubility of 1,8-oxido-p-menthane in volatile oil of magnoliae flos. Using the comprehensive scores of imperatorin and extract yield as indexes,orthogonal test was used to optimize the amount of ethanol,volume fraction and extraction time in extraction technology of Xinzhi nasal drops. Verification test was car-ried out. RESULTS:50% HP-β-CD aqueous solution can improve the solubility of 1,8-oxido-p-menthane to 7.6 times. The optimal extraction technology of Xinzhi nasal drops was as follow as 10-fold 80% ethanol,extracting twice,1 h every time. In verification test,the average content of imperatorin was 0.078%(RSD=2.01%,n=3),and extract yield was 10.80%(RSD=1.85%,n=3). CONCLUSIONS:HP-β-CD shows good solubilization effect on the 1,8-oxido-p-menthane in volatile oil of magnoliae flos,and the optimized extration technology for Xinzhi nasal drops is feasible.

Study on Extraction and Inclusion Processes of Mixed Volatile Oils from Forsythiae Fructus, Saposhnikoviae Radix and Magnoliae Flos / 中国中医药信息杂志

Objective To optimize the extraction process of mixed volatile oil from Forsythiae Fructus, Saposhnikoviae Radix and Magnoliae Flos and inclusion process of β-cyclodextrin (β-CD). Methods With yield ratio of volatile oil as evaluation index, single factor experiments were used to study the extraction process of volatile oil;saturated aqueous solution was used, with inclusion rate of volatile oil as index, and orthogonal design was adopted to examine effects of charge ratio of volatile oil and β-CD, inclusion temperature and inclusion time on the inclusion process; X-ray scattering technique, infrared spectroscopy and scanning electron microscopy were used to characterize the inclusion compound. Results The optimum extraction process of volatile oil was soaking fine powder extracted 5 hours with 10 folds the amount of water. The optimum conditions of inclusion process were as follows:mixed ratio of volatile oil (mL) and β-CD (g) was 1:10; inclusion temperature was 50 ℃; the inclusion time was 2 h. X-ray scattering technique, infrared spectroscopy and scanning electron microscopy proved the inclusion compound had been formed. Conclusion Optimum extraction and inclusion processes are stable and feasible, and can provide research foundation for further research and development of preparation.

Analysis of lignans in Magnoliae Flos by turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry.

In this study, a method coupling turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry was developed for analyzing the lignans in Magnoliae Flos. By the online pretreatment of turbulent flow chromatography solid-phase extraction, the impurities removal and analytes concentration were automatically processed, and the lignans were separated rapidly and well. Seven lignans of Magnoliae Flos including epieudesmin, magnolin, 1-irioresinol-B-dimethyl ether, epi-magnolin, fargesin aschantin, and demethoxyaschantin were identified by comparing their retention behavior, UV spectra, and mass spectra with those of reference substances or literature data. The developed method was validated, and the good results showed that the method was not only automatic and rapid, but also accurate and reliable. The turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry method holds a high potential to become an effective method for the quality control of lignans in Magnoliae Flos and a useful tool for the analysis of other complex mixtures.

Optimization of extraction technology for Xiaochuan Decoction based on information entropy theory / 中草药

Objective: To optimize the extraction technology of Saposhnikoviae Radix, Perillae Folium, Magnoliae Flos, Armeniacae Amarum Semen, and honey-fried Ephedrae Herba in Xiaochuan Decoction by information entropy theory. Methods: With the contents of prem-O-glucosylcimifugin, 4'-O-beta-glucopyranosyl-5-O-methylvisamminol, amygdalin, and the yield of extract as comprehensive evaluation indexes in order to optimize the extraction process parameters of orthogonal test, the weight coefficient of each index was determined by the information entropy weight method. Results: Optimum extraction technology was as follows: reflux extraction for 3 times with 10 fold water, for 1.5 h each time. Conclusion: The optimized method is stable and reliable, and can provide the reference for further development and utilization of the formula.

Pharmacokinetic Study of Magnolin in Rats / 中国药房

OBJECTIVE:To study the pharmacokinetics of magnolin in rats. METHODS:High performance liquid chromatog-raphy(HPLC)was adopted. The determination was performed on Kromasil C18 with mobile phase consisted of acetonitrile-tetrahy-drofuran-water (39∶1∶60),at the flow rate of 1.0 ml/min,with the measurement wavelength of 278 nm,column temperature of 35 ℃ and the sample size of 20 μl. 8 Wistar rats were docked to collect blood from the caudal vein before administration [10 mg(medicinal materials)/kg] and 0.25,0.5,0.75,1,1.5,2,4,8,12 and 20 h after administration,to determine the concentra-tion of the drug in blood. DAS2.1.1 software was used to calculate pharmacokinetic parameters. RESULTS:In the determination of magnolin,the mass concentration linear range was found to be 0.05-10.00 μg/ml(r=0.999 5);the RSD of precision test and sta-bility test were less than 13%(n=6);relative recovery rate was 97.32%-102.15%(n=6);extraction recovery rate was 84.63%-90.02%;t1/2α of magnolin in rats was(0.48±0.22)h,t1/2β was(7.96±2.57)h,CL/F was(0.09±0.032)L/(h·kg),and AUC0-20 h was(944.43±212.83)mg·h/L. CONCLUSIONS:This method conforms to the requirements for the determination of bio-logical samples with respect to precision,stability and accuracy. Magnolin demonstrates a good linear relationship between AUC0-20 h and the dose in rats,with the process compatible with two-compartment model.

Preparation of nasal ethosome sprays of volatile oil in Magnoliae Flos and evaluation of its quality / 中草药

Objective: To study the optimal preparation process of nasal ethosome sprays of volatile oil in Magnoliae Flos after removing a-terpineol alcohol (VOMF) and to investigate its characteristics and mucociliotoxicity. Methods: The 5% VOMF ethosome was prepared with ultrasonic injection method, and the orthogonal test was used to design the formulations of VOMF ethosome which was evaluated by entrapment efficiency and drug loading as indexes. Quality evaluation included appearance, particle diameter distribution, Zeta potential, entrapment efficiency (EE), and stability. The persistent vibration duration (PVD) and percentage of persistent vibration (PPV) in situ toad oral palate cilia were observed to evaluate the mucociliotoxicity administered by various constituents. Results: The optimal preparation was 1.5% of phospholipid, 0.15% of cholesterol, and 36% of ethyl alcohol. The quality evaluation showed that the ethosome was round and uniform, while the mean Zeta potential was (-55.9 ± 2.1) mV. The average particle diameter was getting smaller, the EE had no change, and no mucociliotoxicity was found after spraying. Conclusion: Ultrasonic injection method used to prepare VOMF ethosome is rational and stable and it can be used for nasal administration.

Simultaneous Determination of Four Lignans in Magnoliae Flos Extract by High Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry / 分析化学

A high performance liquid chromatography-electrospray ionization mass spectrometry- charged aerosol detection ( HPLC-MS-CAD) method was established for the simultaneous quantitative analysis of four Lignans in Magnoliae Flos extract. The components were separated on a YMC-Pack ODS-A column (250 mm× 4. 6 mm, 5 μm) by gradient elution with methanol and water as the mobile phase at aflow rate of 1. 0 mL/min. Then the elution solution was routed into MS equipment at a flow rate of 0. 3 mL/min and CAD detector at a flow rate of 0. 7 mL/min by a split ratio of 3:7 for the further detection. The column temperature was 25 ℃ and the detection wavelength was 278 nm. A method was developed for the quantitative analysis of muti-components by single maker ( QAMS) to determine pinoresinol dimethylether, magnoli, 1irioresinol B dimethylethe and epi-magnoli A . Magnoli was selected as internal standard and the relative correction factors ( RCF) of the four Lignans were calculated. The contents of the four Lignans in Magnoliae Flos extract were determined by both external standard method and QAMS. The QAMS method was evaluated by comparison of its assay result and that of external standard method. Under the selected chromatographic condition, the limits of detection of pinoresinol dimethylether, magnoli, lirioresinol B dimethylethe and epi-magnoli A were 0. 34, 0. 55, 0. 50 and 0. 58 mg/L, respectively, while the linear range were within 6. 8-270 mg/L, 11-546 mg/L, 2. 0-101 mg/L and 2. 3-116 mg/L. The recoveries ( n=9 ) were 98. 2%-99. 5%, and the correlation coefficient were 0 . 9995-0 . 9998 . No significant differences were found between the quantitative results of external standard method and QAMS method. The developed method is accurate, feasible, and can be used for quality evaluation of Magnoliae Flos .