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Objective To investigate the effects of honokiol on proliferation and apoptosis of human adipose-derived mesenchymal stem cells(hADSCs),and to investigate the effect of the drug on the tumor microenvironment.Methods hADSCs were incubated with different concentrations of honokiol,the proliferation of hADSCs was detec-ted by MTS and Trypan blue staining,and cell apoptosis was assessed by annexin V/PI double staining.In the meantime,expression of mRNA and protein related to cell proliferation and apoptosis were detected by qPCR and Western blot,respectively.The expression of total MEK,phosphorylated MEK,total ERK and phosphorylated ERK proteins in the MEK-ERK1/2 signaling pathway were detected by Western blot.Results The effect of honokiol on inhibiting proliferation and promoting apoptosis of hADSCs was significantly enhanced with the increase of concen-tration.The expressions of proliferation-related genes CCND1,MKI67 and PCNA were down-regulated.The expres-sions of pro-apoptotic genes BAX and TP53 was up-regulated,and the expressions of anti-apoptotic gene BCL2 was down-regulated.Honokiol inhibited MEK and ERK1/2 phosphorylation in a concentration-dependent manner.Conclusions Honokiol inhibits proliferation and promotes apoptosis of hADSCs,and the specific mechanism is po-tentially related to the inhibition of MEK-ERK1/2 pathway.
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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.
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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.
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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.
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This study was performed to explore the mechanism of myocardial injury attenuation by honokiol via the inhibition of immuno-inflammatory response in ischaemic heart disease(IHD).Twenty of 60 specific-pathogen-free(SPF)male mice were set as sham-operated group,and the remaining mice were subjected to IHD modeling.After model establishment,mice were classified into model group and honokiol group using a random number table method,with 20 mice in each group.Honokiol group was injected intraperitoneally with 0.2 mg/kg of honokiol,and the sham-operated group and model group were given equal volume of 0.9%sodium chloride solution.Cardiac function parameters,including left ventricular end-diastolic pressure(LVEDP),left ventricular systolic pressure(LVSP),maximum rate of left ventricular pressure rise/decline(LV±dp/dtmax),were measured using electrocardiography.Flow cytometry was used to detect the number of Th17/Treg lymphocytes in blood and calculate the Th17/Treg ratio.The levels of IL-1β,IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay.The pathological morphology of myocardium was observed by immunohistochemical staining,the count of myocardial cells was observed under microscope,and the apoptosis rate of myocardial cells was detected by terminal deoxyribonucleotide transferase-mediated dUTP nick end labelling(TUNEL).Compared with sham-operated group,model group and honokiol group demonstrated a reduction in LVSP and LV±dp/dtmax and an increase in LVEDP(P<0.05).Compared with model group,LVSP and LV±dp/dtmax increased and LVEDP decreased in honokiol group(P<0.05).As for Th17/Treg ratio,model group demonstrated the highest level,followed by honokiol group,and sham-operated group showed the lowest level,with statistical difference(all P<0.05).An increase in serum levels of IL-1β,IL-6 and TNF-α were observed in model group and honokiol group when compared to sham-operated group(P<0.05).Levels of IL-1β,IL-6 and TNF-α in serum showed a reduction in honokiol group when compared to model group(P<0.05).The pathological changes including disordered myocardial cell arrangement,severe myocardial necrosis and structural damage,and massive infiltration of inflammatory cells were found in model group and honokiol group,while those changes were alleviated in honokiol group when compared to model group(P<0.05).The cell apoptosis rate showed an increase in model group and honokiol group when compared to sham-operated group(P<0.05),while the cell apoptosis rate was decreased in honokiol group when compared to model group(P<0.05).In conclusion,honokiol pretreatment can improve cardiac function and diminish myocardial injury in IHD.
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Chronic cerebral hypoperfusion leads to white matter injury (WMI), which subsequently causes neurodegeneration and even cognitive impairment. However, due to the lack of treatment specifically for WMI, novel recognized and effective therapeutic strategies are urgently needed. In this study, we found that honokiol and magnolol, two compounds derived from Magnolia officinalis, significantly facilitated the differentiation of primary oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes, with a more prominent effect of the former compound. Moreover, our results demonstrated that honokiol treatment improved myelin injury, induced mature oligodendrocyte protein expression, attenuated cognitive decline, promoted oligodendrocyte regeneration, and inhibited astrocytic activation in the bilateral carotid artery stenosis model. Mechanistically, honokiol increased the phosphorylation of serine/threonine kinase (Akt) and mammalian target of rapamycin (mTOR) by activating cannabinoid receptor 1 during OPC differentiation. Collectively, our study indicates that honokiol might serve as a potential treatment for WMI in chronic cerebral ischemia.
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Magnolia , White Matter , Brain Ischemia/metabolism , Oligodendroglia/metabolismABSTRACT
Aim To investigate the effect of honokiol(HNK)on the angiogenesis of lung cancer cells H460 induced by the tumor inflammatory microenvironment and its possible mechanism.Methods CCK-8 was used to detect the effect of HNK on the proliferation of H460 cells and human umbilical vein endothelial cells(HUVECs); RT-PCR, Western blot, immunofluorescence were used to detect the expression of vascular endothelial growth factor(VEGF); Western blot was used to detect the signaling pathway protein p-IκBα, p-IKKα and NF-κB p65 protein expression.The wound healing test, transwell and tube formation tests were used to detect the inhibition ability of HNK on the migration and angiogenesis of HUVECs.Results HNK treated H460 cells for 24, 48, 72 h respectively, and inhibited the proliferation of H460 cells in a concentration-and time-dependent manner.HNK inhibited the survival of HUVEC cells in a concentration-dependent manner.HNK also reduced the protein and mRNA expression levels of VEGF in H460 cells.Subsequently, HNK concentration-dependently inhibited the phosphorylation of IκBα, NF-κB and IKKα in the NF-κB signaling pathway.Wound healing test, Transwell cell migration test and tube formation test showed that H460 conditioned medium treated with HNK had the ability to inhibit the migration and angiogenesis of HUVECs.Conclusions HNK reduces the viability and angiogenesis of human lung cancer cells by down-regulation of VEGF via the NF-κB pathway.
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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.
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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.
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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.
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Humans , Apoptosis , Biphenyl Compounds/pharmacology , Drugs, Chinese Herbal/pharmacology , Lignans/pharmacology , MagnoliaABSTRACT
Aim To investigate the molecular mechanisms and inhibitory effect of honokiol (HNK) on the proliferation of colon cancer SW620 cells. Methods Crystal violet staining, flow cytometry and Western blot assays were used to for the detection of the effect of HNK on inhibiting the proliferation and promoting apoptosis on SW620 cells. Western blot was used to study the effect of HNK on the protein levels of TGF-β1 and analyze the effect of HNK on the phosphorylation of p38 MAPK, as well as the effect of HNK combined with exogenous adenoviruses of TGF-β1 and its inhibitor on the phosphorylation of p38 MAPK in SW620 cells. Western blot was also used to analyze the effect of HNK on the phosphorylation of YAP and analyze the possible relationship between HNK phosphorylation of p38 MAPK and YAP. Results HNK significantly inhibited the proliferation of SW620 cells and induced apoptosis, and up-regulated the expression levels of TGF-β1. Western blotting showed that HNK up-regu- lated the expression levels of phosphorylated p38 in SW620 cells. Meanwhile, HNK increased the protein levels of phosphorylated YAP. The exogenous adenoviruses of TGF-β1 and its inhibitor significantly enhanced or inhibited this effect, respectively. Conclusions HNK has obvious antiproliferative effect, which might be due to the up-regulation of TGF- (31 expression and up-regulation of phosphorylated YAP expression by activating the p38 MAPK signaling pathway.
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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.
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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.
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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.
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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.
Subject(s)
Humans , Apoptosis , Biphenyl Compounds , Cell Line, Tumor , Cell Proliferation , Lignans , Tongue NeoplasmsABSTRACT
Objective To prepare a honokiol (HNK)-loaded zein nano-drug delivery system with low biotoxicity and to investigate its inhibitory effect on MDA-MB-231 breast cancer cells. Methods Zein-HNK nano-drug delivery system was prepared by anti-solvent method . The particle size‚ dispersion‚ morphological characteristics‚ encapsulation efficiency and in vitro release behavior of the system were investigated. The effects of this system on the proliferation‚ apoptosis‚ migration and invasion of MDA-MB-231 cells were evaluated. Results The particle size of Zein-HNK was (83. 75±2. 95) nm and the polymer dispersity index(PDI) of it was 0. 132± 0. 015. The encapsulation efficiency of HNK was (93. 04± 1. 86)%. The in vitro release rate of Free-HNK group was(88. 90% ± 2. 58)% and release rate of Zein-HNK group was (69. 10± 1. 88)% at 24 hours. Cytotoxicity evaluation showed the 50% inhibiting concentration(IC
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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.
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@#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.
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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.
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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.