Investigation of Pretreatment Methods for Comprehensive Resource Utilization of Magnoliae Officinalis Cortex Dregs / 中国实验方剂学杂志

ObjectiveTo explore the pretreatment methods to promote the enzymatic digestion and extraction of active ingredients from Magnoliae Officinalis Cortex dregs（MOCD）， and to provide a reference basis for the utilization of resource components in MOCD. MethodLiquid chromatography-mass spectrometry（LC-MS） was used for qualitative analysis of resource components in MOCD with an Agilent C18 reversed-phase column（3.0 mm×100 mm， 2.7 µm） at the flow rate of 0.4 mL·min-1， the mobile phase was water（A）-acetonitrile（B） for gradient elution（0-3 min， 25%-48%B； 3-6 min， 48%-59%B； 6-10 min， 59%-80%B； 10-20 min， 80%-90%B； 20-25 min， 90%B）， electrospray ionization（ESI） was employed with negative ion mode scanning and scanning range of m/z 50-1 200. A high performance liquid chromatography（HPLC）， which refered to the determination in the 2020 edition of Chinese Pharmacopoeia， was used for quantitative analysis of resource components in MOCD. Four kinds of pretreatment agents were used to separate the resource components from MOCD， and the mechanism of different pretreatment agents was investigated by field emission scanning electron microscopy（FESEM）， X-ray powder diffraction（XRD） and Fourier transform infrared spectroscopy（FT-IR）. ResultMagnolol， honokiol and lignocellulose were identified as the main resource components of MOCD by qualitative and quantitative analysis. Under the conditions of 1% NaOH， reaction temperature at 80 ℃ and reaction time of 60 min， the concentration of reducing sugar produced by the enzymatic hydrolysis was 32.18 g·L-1， which was 79.8% higher than that of the untreated MOCD. After adding tween-80， the enzymatic hydrolysis time was reduced to 1/3 of the original time， the concentration of reducing sugar was increased by 102.0%. And the total recovery of magnolol and honokiol in the pretreatment solution was 69.23%. ConclusionMagnolol， honokiol and lignocellulosic components in MOCD are valuable for development and utilization， the combination of alkaline pretreatment and tween-80 can realize the recovery and utilization of these three resource components， which can provide a new idea for comprehensive utilization of resource components in MOCD.

Honokiol alleviated neurodegeneration by reducing oxidative stress and improving mitochondrial function in mutant SOD1 cellular and mouse models of amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease affecting both upper and lower motor neurons (MNs) with large unmet medical needs. Multiple pathological mechanisms are considered to contribute to the progression of ALS, including neuronal oxidative stress and mitochondrial dysfunction. Honokiol (HNK) has been reported to exert therapeutic effects in several neurologic disease models including ischemia stroke, Alzheimer's disease and Parkinson's disease. Here we found that honokiol also exhibited protective effects in ALS disease models both in vitro and in vivo. Honokiol improved the viability of NSC-34 motor neuron-like cells that expressed the mutant G93A SOD1 proteins (SOD1-G93A cells for short). Mechanistical studies revealed that honokiol alleviated cellular oxidative stress by enhancing glutathione (GSH) synthesis and activating the nuclear factor erythroid 2-related factor 2 (NRF2)-antioxidant response element (ARE) pathway. Also, honokiol improved both mitochondrial function and morphology via fine-tuning mitochondrial dynamics in SOD1-G93A cells. Importantly, honokiol extended the lifespan of the SOD1-G93A transgenic mice and improved the motor function. The improvement of antioxidant capacity and mitochondrial function was further confirmed in the spinal cord and gastrocnemius muscle in mice. Overall, honokiol showed promising preclinical potential as a multiple target drug for ALS treatment.

Honokiol attenuates acetaminophen-induced acute liver injury by inhibiting hepatic CYP1A2 activity and improving liver mitochondrial dysfunction / 中草药·英文版

OBJECTIVE@#Acetaminophen (APAP) overdose is a common cause of liver injury. This study aimed to investigate the protective effect of honokiol (Hon) against APAP-induced hepatotoxicity and its potential mechanism.@*METHODS@#C57BL/6 mice were administrated with Hon (10 and 30 mg/kg) after APAP (300 mg/kg) treatment. On 1.5 h and 5 h after Hon treatment, mice were sacrificed. Serum and liver were collected. And then, liver injury-related indexes, APAP metabolism-related indexes, mitochondrial respiratory chain function-related indexes, and mitochondrial membrane function-related protein expression were evaluated.@*RESULTS@#It was found that Hon significantly decreased serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) activity and glutathione (GSH) depletion, increased hepatic catalase (CAT) and GSH peroxidase (GSH-Px) activities, reduced hepatic MDA and 3-nitrotyrosine contents, inhibited hepatic CYP1A2 activity and APAP protein adducts (APAP-CYS) formation. Meanwhile, oxidative phosphorylation capacity of complex I and electron transfer capacity of complex IV in mitochondrial respiratory chain was increased, whereas the release of H2O2 in the mitochondria was decreased following Hon treatment. Furthermore, Hon markedly down-regulated p-JNK in both cytosol and mitochondria, and obviously inhibited the release of apoptosis inducing factor (AIF) and endonuclease G (EndoG) from mitochondria to cytosol.@*CONCLUSION@#Hon alleviated APAP-induced liver injury through the following pathways: Reducing the production of APAP-CYS by inhibiting CYP1A2 activity; Ameliorating hepatic oxidative stress by increasing the levels of hepatic CAT, GSH-Px and GSH; Improving mitochondrial respiratory chain function by promoting oxidative phosphorylation capacity of complex I and electron transfer capacity of complex IV; Improving the function of mitochondrial membrane by inhibiting p-JNK and its translocation to mitochondria, thereby reducing the release of AIF and EndoG.

Honokiol: A naturally occurring lignan with pleiotropic bioactivities / 中国天然药物

Honokiol is the dominant biphenolic compound isolated from the Magnolia tree, and has long been considered as the active constituent of the traditional Chinese herb, 'Houpo', which is widely used to treat symptoms due to 'stagnation of qi'. Pharmacological studies have shown that honokiol possesses a wide range of bioactivities without obvious toxicity. Honokiol protects the liver, kidneys, nervous system, and cardiovascular system through reducing oxidative stress and relieving inflammation. Moreover, honokiol shows anti-diabetic property through enhancing insulin sensitivity, and anti-obese property through promoting browning of adipocytes. In vivo and in vitro studies indicated that honokiol functions as an anti-cancer agent through multiple mechanisms: inhibiting angiogenesis, promoting cell apoptosis, and regulating cell cycle. A variety of therapeutic effects of honokiol may be associated with its physiochemical properties, which make honokiol readily cross the blood brain barrier and the blood-cerebrospinal fluid barrier, with high bioavailability. In the future, more clinical researches on honokiol are needed to fully authenticate its therapeutic values.

Honokiol attenuates oxidative stress-induced cytotoxicity in human keratinocytes via activating AMPK signaling

Objective: To investigate the effect of honokiol on oxidative damage in HaCaT human keratinocytes. Methods: HaCaT cells were exposed to hydrogen peroxide (H2O2), following pretreatment with various concentrations of honokiol. The alleviating effects of honokiol on HaCaT cell viability and cell death, reactive oxygen species (ROS) production, DNA damage, mitochondrial dynamics, and inhibition of adenosine triphoaphate production against H2O2 were investigated. Western blotting analysis was used to analyze the expression levels of specific proteins. Results: Honokiol suppressed H2O2-induced cytotoxicity and DNA damage by blocking abnormal ROS accumulation. Honokiol also prevented apoptosis by inhibiting loss of mitochondrial membrane potential and release of cytochrome c from the mitochondria into the cytosol, decreasing the Bax/Bcl-2 ratio, and reducing the activity of caspase-3 in H2O2-stimulated HaCaT cells. In addition, honokiol attenuated H2O2-induced reduction of adenosine triphosphate content, and activation of AMP-activated protein kinase (AMPK) was markedly promoted by honokiol in H2O2-stimulated cells. Importantly, the anti-apoptosis and anti-proliferative activity of honokiol against H2O2 was further enhanced by adding an activator of AMPK, indicating that honokiol activated AMPK in HaCaT keratinocytes to protect against oxidative damage. Conclusions: The present results indicate that honokiol may be useful as a potential therapeutic agent against various oxidative stress-related skin diseases.

Honokiol attenuates oxidative stress-induced cytotoxicity in human keratinocytes via activating AMPK signaling

Objective: To investigate the effect of honokiol on oxidative damage in HaCaT human keratinocytes. Methods: HaCaT cells were exposed to hydrogen peroxide (H2O2), following pretreatment with various concentrations of honokiol. The alleviating effects of honokiol on HaCaT cell viability and cell death, reactive oxygen species (ROS) production, DNA damage, mitochondrial dynamics, and inhibition of adenosine triphoaphate production against H2O2 were investigated. Western blotting analysis was used to analyze the expression levels of specific proteins. Results: Honokiol suppressed H2O2-induced cytotoxicity and DNA damage by blocking abnormal ROS accumulation. Honokiol also prevented apoptosis by inhibiting loss of mitochondrial membrane potential and release of cytochrome c from the mitochondria into the cytosol, decreasing the Bax/Bcl-2 ratio, and reducing the activity of caspase-3 in H2O2-stimulated HaCaT cells. In addition, honokiol attenuated H2O2-induced reduction of adenosine triphosphate content, and activation of AMP-activated protein kinase (AMPK) was markedly promoted by honokiol in H2O2-stimulated cells. Importantly, the anti-apoptosis and anti-proliferative activity of honokiol against H2O2 was further enhanced by adding an activator of AMPK, indicating that honokiol activated AMPK in HaCaT keratinocytes to protect against oxidative damage. Conclusions: The present results indicate that honokiol may be useful as a potential therapeutic agent against various oxidative stress-related skin diseases.

Optimization of supercritical CO2 extraction process of Magnoliae Officinalis Cortex and anti-oxidant activities of extracts / 中草药

Objective: To optimize the supercritical CO2 extraction process of active ingredients from Magnoliae Officinalis Cortex (MOC) and explore the antioxidant activity of the extracts. Methods: The content of magnolol and honokiol of the supercritical CO2 extracts of MOC was determined by HPLC, and the extraction process was optimized by orthogonal experiment. The antioxidant activities of the extracts were determined by MTT. Results: The optimum extraction pressure of magnolol was 25 MPa, the extraction temperature was 55 ℃, the amount of CO2 was 30 kg, and the optimum extraction parameters mentioned above of honokiol were 15 MPa, 50 ℃, and 25 kg, respectively. Conclusion: Under the optimum extraction conditions, magnolol and honokiol have high extraction efficiency, good repeatability, stability and feasibility, and the extract have good antioxidant activity.

Study on quality transfer law of Magnoliae Officinalis Cortex in processing based on nine components determination / 中草药

Objective: To reveal the scientific connotation of ginger-processed Magnoliae Officinalis Cortex (MOC), and standardize the production process and offer the theoretical base for clinical medication. Methods: This paper studied on quality transfer law of MOC in processing based on the determination of nine chemical components, including syringing, magnocurarine, magnolin B, magnoflorine, magnolin A, honokiol, magnolol, piperitylmagnolol, and β-eudesmol, through controlling factors such as part of sample collection, processing technology, personnel, equipment and environment. Results: The results showed that the content of phenolic components was increased slightly, the content of alkaloids was decreased significantly, and the content of glycosides was decreased, and the content of magnoloside B was decreased significantly in the process of preparation for MOC and ginger-processed MOC. Moreover, there was no significant difference in the content of magnoloside A, syringin, and volatile oil represented by β-eudesmol. Conclusion: This study preliminarily explored the quality transfer law of multi-components in the processing of MOC, and provided reference for the establishment of the quality control system of the crude drug processing technology and improvement of the quality of products.

Effect of honokiol on proliferation, migration and apoptosis of human tongue cancer CAL-27 cells / 南方医科大学学报

OBJECTIVE@#To study the effects of honokiol on proliferation, migration and apoptosis of human tongue carcinoma CAL-27 cells.@*METHODS@#Routinely cultured CAL-27 cells were treated with 20, 40, or 60 μmol/L honokiol and the changes in cell proliferation were assessed with MTT assay. The scratch wound healing assay was used to assess the migration ability of the treated cells, and the cell apoptosis was detected with Hoechst33342 fluorescence staining and annexin V-FITC/PI method. The protein expression levels of p-Pi3k, p-Fak, Fak, MMP-2, MMP-9, p-Akt, Akt, Bax, Bcl-2 and cleaved-caspase-3 in the treated cells were detected using Western blotting.@*RESULTS@#Treatment with honokiol at 20, 40, and 60 μmol/L for 24 h significantly lowered the proliferation and migration ability of CAL-27 cells. The number of apoptotic cells increased with the increase of honokiol concentration, which resulted in a cell apoptosis rate of (15.24±2.06)% at 20 μmol/L, (35.03±2.42)% at 40 μmol/L, and (48.13±4.61)% at 60 μmol/L, as compared with (6.53±1.80)% in the control group. The expressions of p-Pi3k, p-Fak, MMP-2, MMP-9, p-Akt and BCL-2 decreased and those of Bax and cleaved-caspase-3 increased significantly in the cells after the treatment ( < 0.01).@*CONCLUSIONS@#Honokiol can inhibit the proliferation and migration and induce apoptosis of CAL-27 cells possibly by regulating the expressions of p-Pi3k, p-Fak, MMP-2, MMP-9, p-Akt, Bax, Bcl-2 and cleaved-caspase-3.

Research Progress in Modern Pharmacology of Magnolia Officinalis / 中国实验方剂学杂志

Magnoliae Officinalis Cortex（MOC） is a commonly used traditional Chinese medicine（TCM） in China. It is spicy-warm in property and bitter in flavor. It has the effects in eliminating dampness， eliminating phlegm and removing fullness. It is commonly used for dampness obstruction to spleen and stomach， chest and epigastric distension， glutinous grains， nausea， vomiting， abdominal pain and abdominal distension. It has a good efficacy in treating gastrointestinal discomfort and anorexia in clinic. The results showed that MOC mainly contains phenolic compounds， alkaloids and volatile oil. Magnolol， honokiol and other phenolic compounds are the main active substances， with obvious pharmacological activities on digestive， nervous， cardiovascular and respiratory systems. In addition， it also has anti-inflammatory， analgesic， anti-bacterial， anti-tumor and anti-oxidation effects. Except for magnolol and honokiol and other active substances， MOC flowers also contain volatile oil， with a similar effect with MOC but a weaker function. It is mainly used for treating spleen and stomach dampness， fullness， chest and epigastric distension. In addition to magnolol and honokiol and other phenolic compounds， MOC leaves also contain volatile oil， flavonoids and polysaccharides and other chemical components， which have antibacterial， antioxidative， vasodilatory and other pharmacological effects. It can be used as medicine instead of MOC in clinic. In this paper， the pharmacology studies of MOC in recent 5 years was reviewed， in order to better develop and utilize magnolia bark and its waste flowers and leaves， and further develop relevant functional products with MOC as the main drug， while providing new ideas for expanding the resources of TCM.

Beneficial effects of nanosuspensions of honokiol in mice on high fat diet through suppression of hepatic gluconeogenesis / 药学学报

To investigate the potential hypoglycemic effect of nanosuspensions of honokiol and explore the underlying mechanisms, a high fat diet (HFD) was studied in C57BL/6J mice divided into five groups: normal diet (ND), HFD, HFD/honokiol-sodium carboxymethyl cellulose (CMC-Na) (Hono-CMC, 100 mg·kg-1), HFD/honokiol- Nano (Hono-Nano, 80 mg·kg-1), HFD/metformin (HFD/Met, 200 mg·kg-1). Fasting blood glucose (FBG) and body weights (BW) of mice were measured every seven days. After 30-day treatment, an oral glucose tolerance test (OGTT) was performed, and blood and tissue samples were collected for analysis. All animal experiments were approved by the Research Animal Care Committee of Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine. The data showed Hono-Nano and metformin reduced FBG, BW, and markedly improved OGTT of mice compared to HFD group (P<0.05). Hono-CMC produced nonsignificant impact on FBG, BW of mice, while OGTT of mice was improved by Hono-CMC (P<0.05). Meanwhile, none of these treated groups showed significant effects on regulating serum insulin levels, but all of them exhibited decreased serum glucagon levels notably compared to the HFD group (P<0.05). Western blot analysis revealed that honokiol up-regulated levels of p-AMPK and p-FOXO1 in liver tissue of HFD mice (P<0.05), which resulted in activation of AMPK and inhibition of FOXO1. Moreover, the expression of PEPCK (a key enzyme of gluconeogenesis) was decreased by honokiol (P<0.05). Taken together, our findings demonstrate that nanosuspension of honokiol is more effective than CMC-Na-suspension of honokiol on blood glucose controlling in HFD mice. The hypoglycemic effects of honokiol might rely on suppressing hepatic gluconeogenesis via activating AMPK and inhibiting FOXO1.

Grade Evaluation of Magnoliae Officinalis Cortex Processed with Ginger Juice Based on Relative Quality Constant / 中国实验方剂学杂志

Objective: To establish the grade evaluation standard for Magnoliae Officinalis Cortex processed with ginger juice by combining traditional morphology evaluation with modern intrinsic quality evaluation. Method: The morphological parameters and contents of intrinsic pharmacodynamic index components of 28 batches of Magnoliae Officinalis Cortex processed with ginger juice were determined, and the relative quality constants were calculated. Assuming that the average relative quality constant was 100%, more than 120%of the samples were classified as the first grade, 60%to 120%as the second grade, the remaining as the third grade. Result: The relative quality constant of Magnoliae Officinalis Cortex processed with ginger juice ranged from 0.09 to 1.78. The relative quality constant of the first grade was ≥ 0.64, the second grade was 0.32-0.64, while the third grade was ≤ 0.32. Conclusion: Relative quality constant combines external indexes of traditional morphology and internal indexes of pharmacodynamic components, which can objectively, reasonably and scientifically classify the grade of Magnoliae Officinalis Cortex processed with ginger juice, and provide reference for establishing and improving the grade evaluation standard of this decoction pieces.

Surface solid dispersion of honokiol-croscarmellose sodium:preparation and characterization / 中国药科大学学报

@#The surface solid dispersion of honokiol was prepared by melting method with croscarmellose sodium as carrier to improve the dissolution rate of honokiol, taking the advantage of its low melting point. The dissolution rate and stability test of surface solid dispersion of honokiol were carried out. Its physical properties were then investigated by DSC, PXRD and SEM analysis. The results indicated that the dissolution rate of honokiol from sodium solid dispersion was more than 90% at 15 min while its mean dissolution time was significantly decreased. Honokiol was distributed on the surface of croscarmellose sodium in the form of microcrystal; moreover, its dissolution behavior didn′t change significantly after six months of stability tests. Therefore, surface solid dispersion of honokionl could be prepared by simple process. The formed solid dispersion achieved high drug loading and fast in vitro dissolution rate, which could provide a novel idea for developing solid preparations of honokiol.

Research on standard decoction of ginger juice Magnoliae Officinalis Cortex / 中草药

Objective To study the dry extract rate, determination and transfer rate of maker compounds, and fingerprint of standard decoction of ginger juice Magnoliae Officinalis Cortex (GJMOC) and provide a reference for the preparation and quality assessment of its dispensing granules by establishing 16 batches of standard decoction of GJMOC. Methods A total of 16 batches of GJMOC standard decoctions were prepared following literature requirements. The quantitative analysis method of magnolol and honokiol was according to Chinese Pharmacopoeia (2015 edition). The transfer rate of total magnolol and honokiol and extraction rate were calculated. the pH value was determined and HPLC fingerprint was established under a flow rate of 1 mL/min and eluted with a mobile phase of acetonitrile (A)-0.1% phosphoric acid solution (B) in a gradient mode (0-15 min, 12%-16% A; 15-30 min, 16%-28% A; 30-42 min, 28%-74% A; 42-55 min, 74%-80% A). The column temperature was set at 40℃ and the detection wavelength was 294 nm. Results By measuring the of 16 batches of standard decoction, the transfer rate of the sum of magnolol and honokiol ranged from 6.5% to 12.0%, the extraction rate was at a range of 3.41% to 7.14% and pH value was 4.63 to 5.43. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012A) was used to analyze and compare the fingerprints, and seven common peaks were determined and four were identified including magnoloside B (peak 1), magnoloside A (peak 2), honokiol (peak 6), and magnolol (peak 7). The similarity among 16 batches of standard decoction of GJMOC was evaluated, and the similarity was all greater than 0.69. Moreover, this study established an HPLC fingerprint analysis method of GJMOC standard decoction. Conclusion The preparation method established in this study is stable and feasible, and the analysis method shows good precision, stability, and repeatability in fingerprint analysis and it is suitable for evaluating the quality of standard decoction of GJMOC.

Effect of honokiol on PI3K/Akt signaling pathway in asthmatic mice and its effect on TLR2 and TLR4 expression / 中草药

Objective: To investigate the effect of honokiol on PI3K/Akt signaling pathway in asthmatic mice and its effect on TLR2 and TLR4 expression. Methods Fifty ICR mice were randomly divided into normal control group, asthma model group, positive control group (dexamethasone 10 mg/kg, ip), and honokiol high and low dose (50 μg/kg and 10 μg/kg, ip) groups. The asthma model was induced by ovalbumin plus aluminum hydroxide sensitization method, and then the corresponding drugs were administered for 7 d. The alveolar lavage fluid was collected and white blood cells were counted at 24 h after the last administration; The histopathological examinations of HE, PAS and Masson staining were performed. The expression of PI3K, Akt, p-Akt, TLR2, and TLR4 protein in lung tissue was detected by Western blotting. The mRNA expression of TLR2 and TLR4 in lung tissue of mice was detected by RT-PCR. Results Compared with the asthma model group, the numbers of neutrophil, lymphocyte and eosinophil in the alveolar lavage fluid of mice treated with honokiol 50 μg/kg group were significantly reduced (P < 0.01); Lung pathology was significantly improved, and inflammatory cell infiltration was significantly reduced (P < 0.05). The tracheal wall and alveolar damage were significantly relieved; The expression levels of PI3K, Akt, p-Akt, TLR2, TLR4 protein and TLR2, TLR4 mRNA were significantly down-regulated in lung tissue (P < 0.05). Conclusion Honokiol has a good inhibitory effect on OVA-induced inflammatory response in asthmatic mice. The mechanism may be related to the inhibition of PI3K/Akt signaling pathway by TLR2 and TLR4 receptors.

Effects of honokiol on activation of transient receptor potential channel in rat spinal dorsal root ganglion cells and pruritus in mouse models / 中华皮肤科杂志

Objective To evaluate the effect of honokiol on the activation of transient receptor potential (TRP) channel in rat spinal dorsal root ganglion cells and pruritus in mouse models.Methods Healthy male ICR mice aged 4-6 weeks were used to establish histamine-induced and acetone/ether/water (AEW)-induced itching models separately.Totally,mice were randomly divided into 7 groups (histamineinduced model experiment) or 6 groups (AEW-induced model experiment):normal control group and model group both gavaged with sodium chloride physiological solution,solvent group gavaged with sodium carboxymethylcellulose solution,chlorphenamine group (only set up in the histamine-induced model experiment) gavaged with chlorphenamine,50,25 and 12.5 mg/kg honokiol groups gavaged with 50,25 and 12.5 mg/kg honokiol respectively.In the histamine-induced model experiment,the mice were all injected with histamine except the normal control group injected with sodium chloride physiological solution 24 hours after the gavage treatment,while the mice in the AEW-induced model experiment were all topically treated with AEW except the normal control group topically treated with sodium chloride physiological solution for 4 days,followed by gavage with different drugs.The anti-itch effect of each treatment was evaluated by counting the scratching frequency within 30 minutes.Rat spinal dorsal root ganglion (DRG)cells were isolated and subjected to a primary culture.Then,the DRG cells were divided into 6 groups:capsaicin or allyl isothiocyanate (AITC)-induced model group pre-incubated with Hank's balanced salt solution (HBSS),500 μmol/L capsazepine or 10 μmol/L HSC030031 group pre-incubated with capsazepine or HSC030031,solvent group pre-incubated with dimethyl sulfoxide (DMSO),3 honokiol groups preincubated with 7.81,15.63 and 31.25 mg/L honokiol respectively,and Ca2+ fluorescence imaging system was used to observe changes of Ca2+ influx in these cells after capsaicin or AITC stimulation.Statistical analysis was carried out with SPSS 20.0 software by using one-way analysis of variance and Dunnett-t test.Results In the histamine-induced mouse models,the scratching frequency was significantly lower in the 50 and 25 mg/kg honokiol groups than in the model group (21.88 and 21.14 vs.63.70,t =3.48,3.49 respectively,both P =0.003),while no significant difference in the scratching frequency was observed between the 12.5 mg/kg honokiol group and the model group (t =2.01,P =0.062).After the treatment with 50 mg/kg honokiol in the AEW-induced mouse models,the scratching frequency significantly decreased compared with the model group (61.4 vs.101.17,t =0.45,P =0.009),while there were no significant differences among the 25,12.5 mg/kg honokiol groups and the model group (all P ＞ 0.05).Compared with the capsaicin or AITC-induced model group,the increase of Ca2+ fluorescence signal in the DRG cells was significantly inhibited in the 31.25 mg/L honokiol group:at the 45th second,the rate of relative fluorescence intensity change (AF/F0) was 1.11 in the model group,but-0.11 in the 31.25 mg/L honokiol group in the capsaicin-induced model experiment,and 0.56 in the model group,but 0.00 in the 31.25 mg/L honokiol group in the AITC-induced model experiment.Conclusion Honokiol shows an inhibitory effect on mouse models of pruritus induced by histaminergic or non-histaminergic factors,likely by inhibiting Ca2+ influx through activated TRPV 1 and TRPA 1 channels in the DRG cells.

The effects of honokiol on lipid synthesis in HepG2 cells with steatosis / 药学学报

In this study, the effects of honokiol (HN) treatment for 24 h on lipid synthesis was examined in HepG2 cells. The parameters include intracellular lipid droplet and the expression of SREBP-1c and PNPLA3, glucose uptake, and oxidative stress including the expression of CYP2E1 and CYP4A in normal, TO901317 (TO)- and oleic acid (OA)-treated HepG2 cells. The lipid droplets were detected by oil red O staining. The glucose uptake was measured by fluorescence spectrophotometry using[2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose, 2-NBDG] as probe. The expression levels of target genes were detected by quantitative PCR and Western blot. The results showed that:① TO (5 μmol·L−1) and OA (0.5 mmol·L−1) treatment increased the levels of intracellular lipid accumulation and the mRNA and protein expression of SREBP-1c and PNPLA3. After HN (10, 20, 40 μmol·L−1) treatment for 24 h, the lipid accumulation and the expression of SREBP-1c and PNPLA3 were all decreased in the tested cells. ② OA treatment significantly suppressed glucose uptake, while HN treatment dose-dependently increased the glucose uptake in OA-treated cells. ③ Compared with control group, CYP2E1 protein level significantly decreased in the three tested cells, and CYP4A protein level significantly decreased only in OA-treated cells following HN treatment. The above results suggest that HN may attenuate lipid accumulation by suppressing the expression of SREBP-1c and PNPLA3, and reduce lipid peroxidation and insulin resistance by down-regulation of the protein levels of CYP2E1 and CYP4A in HepG2 cells with steatosis.

Comparison study on the pharmacokinetic and tissue distribution of honokiol and its metabolites between normal and diabetic rats / 药学学报

This study was designed to compare normal and diabetic rats in the pharmacokinetic and tissue distribution of honokiol and its metabolites. Type 2 diabetic rat model was established using high-fat diet feeding for 6 weeks followed by single intraperitoneal injection of low dose (30 mg·kg-1) strepotozotocin. The concentration of honokiol and its metabolites were determined by high performance liquid chromatography. Pharmacokinetic parameters were calculated using DAS 3.0 software. The Cmax of honokiol in the normal rats and diabetic rats are 872.5 ±233.1 and 614.7 ±182.4 μg·L-1 respectively (P -1 respectively. Meanwhile, AUC0-t and Cmax of the metabolite M2 were significantly increased in diabetic rats. The concentrations of honokiol in liver, kidney and brain in the diabetic model group were higher than those of the normal control group. Meanwhile, the exposure of M1 in liver, M2 in liver and kidney of the model group were higher than that in the normal group. The data suggest that the pharmacokinetics and tissue distribution of honokiol and its metabolites are changed in the pathological state of diabetes.

Honokiol nanosuspensions:preparation, in vitro and in vivo evaluation / 药学学报

Honokiol (HK) have extensive pharmacological activities, but its poor solubility and instability restricted its clinical application and efficacy exertion. HK nanosuspensions (HK-NSps) were designed in this study in order to solve the problems. HK-NSps were prepared by antisolvent precipitation method, using poly-vinylpyrrolidone (PVP) and bovine serum albumin (BSA) as a combined stabilizer. The particle size was measured using dynamic light scattering method, the morphology was observed by transmission electron microscopy. The size change and drug content of HK-NSps in various physiological media during the storage at ambient temperature was examined to evaluate their storage stability. Dialysis method was used to study their drug release in vitro. MTT assay was used to assess their in vitro cytotoxicity against 4T1 breast cancer cell line. Anti-tumor effect in vivo was also investigated in 4T1 tumor-bearing mice. HK-NSps were prepared with high drug loading content of 48.62%, nearly spherical shape and good storage stability. The average particle size was (83.40 ±1.042) nm, the polydispersity index (PDI) value was 0.223 ±0.011, the zeta potential was (-42.2 ±1.2) mV. HK-NSps showed sustained in vitro drug release and enhanced cytotoxicity in contrast to free HK against 4T1 cells (IC50, 8.36 μg·mL-1 vs 37.58 μg·mL-1, Pin vivo study on 4T1 tumor-bearing mice demonstrated that HK-NSps showed good dose-dependent tumor inhibition rate (TIR). In contrast to 4 mg·kg-1 of PTX injection (TIR, 47.9%), medium and high dose of HK-NSps displayed improved therapeutic efficacy (TIR, 55.67% for 40 mg·kg-1, 67.28% for 60 mg·kg-1, P-1) had TIR of only 54.13% even administrated every day. In conclusion, HK-NSps were prepared with small size, high drug-loading capacity, and good stability. The improved in vitro and in vivo antitumor efficacy demonstrated that HK can be a promising antitumor drug in combination with nanosuspensions technology.

Simultaneous Determination of Six Effective Components in Dutong Pills by GC / 中国药师

Objective:To establish a GC method for the imultaneous determination of cinnamaldehyde, bornyl acetate, costunol-ide,dehydrocostus lactone,magnolol and honokiol in Dutong pills. Methods: The determination was performed on an HP-5 column (30 m ×0.32 mm,0.25 μm) with programmed temperature. The carrier gas was nitrogen with the flow rate of 2.0 ml·min-1. The injection volumn was 1 μl and the sample split ratio was 5:1. The inlet temperature was 280 ℃. The detector was a flame ionization detector with temperature at 300 ℃. Results:The linear ranges were 32.28-516.40 μg·ml-1for cinnamaldehyde(r=0.999 3), 27.06-433.00 μg·ml-1for bornyl acetate(r=0.999 2),25.65-410.40 μg·ml-1for costunolide(r=0.999 3),26.10-417.60 μg ·ml-1for dehydrocostus lactone(r=0.999 3),24.01-384.20 μg·ml-1for magnolol(r=0.999 0) and 18.32-293.10 μg·ml-1 for honokiol(r=0.999 4). The average recovery was 99.71%(RSD=0.67%),99.34%(RSD=1.18%),100.16%(RSD=0. 34%),100.40%(RSD=0.39%),99.32%(RSD=1.22%) and 99.58%(RSD=0.58%)(n=6),respectively. Conclusion:The method is simple,sensitive and accurate,can be used to supplement the insufficient quality control of Dutong pills.