Effect of honokiol regulating SIRT3 on chronic hypoxia-induced microglia and astrocyte polarization

Objective:To investigate the effect of honokiol on microglia polarization and the underlying mechanism.Methods:Inflammatory factors were detected using ELISA to determine the optimal concentration of cobalt chloride to induce,and that of honokiol to treat chronic hypoxia(48 h)in microglia cell line BV2 cells.BV2 cells were divided into four groups:control,chronic hypoxia,chronic hypoxia+honokiol,chronic hypoxia+honokiol+3-TYP(SIRT3 inhibitor).ELISA was used to measure the concentration of supernatant TNFαand IL-1βproteins,qPCR was used to detect the expression of cellular M1 and M2 polarization markers,and biochemical assays were used to detect the level of reactive oxygen species in each group.Western Blot was used to detect protein levels of SIRT3 and upstream inflammatory molecules NLRP3 and caspase1.Results:Chronic cobalt chloride stimulation of BV2 cells at an optimal concentration of 100μmol/L significantly increased the release of inflammatory fac-tors TNFαand IL-1βafter stimulation compared with the control group(P<0.05);compared with the control group,cells in the chronic hypoxia group had down-regulation of SIRT3 protein expression,whereas the ROS levels,NLRP3 and caspase1 protein levels,the M1 polarization marker CD86,iNOS mRNA levels and CD16/32 ratio were upregulated.and honokiol(10μmol/L)significantly up-regulated the SIRT3 protein and mRNA levels of M2 markers Arg-1 and CD206 in chronic hypoxic cells(P<0.05)and down-regulated levels of ROS,NLRP3/caspase1 protein,and mRNA levels of M1 markers(P<0.05),and this anti-oxidative and anti-inflammatory effect was able to be reversed by SIRT3 inhibitor.Conclusion:Honokiol inhibits chronic hypoxia-induced microglia M1 polarization and inflammatory pathway activation,and its anti-inflammatory effects are SIRT3-de-pendent.

Honokiol:a promising small molecular weight natural agent for the growth inhibition of oral squamous cell carcinoma cells

Honokiol （HNK） is a small organic molecule purified from magnolia species and has demonstrated anticancer activities in a variety of cancer cell lines; however, its effect on oral squamous cell carcinoma （OSCC） cells is unknown. We investigated the antitumor activities of HNK on OSCC ceils in vitro for the first time. The inhibitory effects of HNK on the growth and proliferation of OSCC cells were demonstrated via in vitro 3-（4,5-dimethyl thiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） and propidium iodide （PI） assays, and the apoptotic cells were investigated by the observation of morphological changes and detection of DNA fragmentation via PI, TdT-mediated dUTP-biotin nick end labeling （TUNEL）, and DNA ladder assays, as well as flow cytometry assay. The results showed that HNK inhibited the growth and proliferation of OSCC cells in vitro in a time and dose-dependent manner. The inhibitory effect was associated with the cell apoptosis induced by HNK, evidenced by the morphological features of apoptotic cells, TUNEL-positive cells and a degradation of chromosomal DNA into small internucleosomal fragments. The study also demonstrated here that the inhibition or apoptosis mediated by 15 μg.mL-1 or 20 μg.mL-1 of HNK were more stronger compared with those of 20 μg-mL-1 5-fluorouracil （5-Fu, the control） applied to OSCC cells, when the ratio of OSCC cell numbers were measured between the treatment of different concentrations of HNK to the 5-Fu treatment for 48 h. HNK is a promising compound that can be potentially used as a novel treatment agent for human OSCC.

Supplemental magnolol or honokiol attenuates adverse effects in broilers infected with Salmonella pullorum by modulating mucosal gene expression and the gut microbiota

Background:Salmonella pullorum is one of the most harmful pathogens to avian species.Magnolol and honokiol,natural compounds extracted from Magnolia officinalis,exerts anti-inflammatory,anti-oxidant and antibacterial activities.This study was conducted to evaluate the effects of dietary supplemental magnolol and honokiol in broilers infected with S.pullorum.A total of 360 one-day-old broilers were selected and randomly divided into four groups with six replicates:the negative control group(CTL),S.pullorum-infected group(SP),and the S.pulloruminfected group supplemented with 300 mg/kg honokiol(SPH)or magnolol(SPM).Results:The results showed that challenging with S.pullorum impaired growth performance in broilers,as indicated by the observed decreases in body weight(P<0.05)and average daily gains(P<0.05),along with increased spleen(P<0.01)and bursa of Fabricus weights(P<0.05),serum globulin contents,and the decreased intestine villus height and villus/crypt ratios(P<0.05).Notably,supplemental magnolol and honokiol attenuated these adverse changes,and the effects of magnolol were better than those of honokiol.Therefore,we performed RNA-Seq in ileum tissues and 16S rRNA gene sequencing of ileum bacteria.Our analysis revealed that magnolol increased the α-diversity(observed species,Chao1,ACE,and PD whole tree)and β-diversity of the ileum bacteria(P<0.05).In addition,magnolol supplementation increased the abundance of Lactobacillus(P<0.01)and decreased unidentified Cyanobacteria(P<0.05)both at d 14 and d 21.Further study confirmed that differentially expressed genes induced by magnolol and honokiol supplementation enriched in cytokine-cytokine receptor interactions,in the intestinal immune network for IgA production,and in the cell adhesion molecule pathways.Conclusions:Supplemental magnolol and honokiol alleviated S.pullorum-induced impairments in growth performance,and the effect of magnolol was better than that of honokiol,which could be partially due to magnolol’s ability to improve the intestinal microbial and mucosal barrier.

Honokiol-enhanced cytotoxic T lymphocyte activity against cholangiocarcinoma cells mediated by dendritic cells pulsed with damage-associated molecular patterns

BACKGROUND Cholangiocarcinoma or biliary tract cancer has a high mortality rate resulting from late presentation and ineffective treatment strategy. Since immunotherapy by dendritic cells (DC) may be beneficial for cholangiocarcinoma treatment but their efficacy against cholangiocarcinoma was low. We suggest how such antitumor activity can be increased using cell lysates derived from an honokioltreated cholangiocarcinoma cell line (KKU-213L5). AIM To increase antitumour activity of DCs pulsed with cell lysates derived from honokiol-treated cholangiocarcinoma cell line (KKU-213L5). METHODS The effect of honokiol, a phenolic compound isolated from Magnolia officinalis, on choangiocarcinoma cells was investigated in terms of the cytotoxicity and the expression of damage-associated molecular patterns (DAMPs). DCs were loaded with tumour cell lysates derived from honokiol-treated cholangiocarcinoma cells their efficacy including induction of T lymphocyte proliferation, proinflammatory cytokine production and cytotoxicity effect on target cholangiocarcinoma cells were evaluated. RESULTS Honokiol can effectively activate cholangiocarcinoma apoptosis and increase the release of damage-associated molecular patterns. DCs loaded with cell lysates derived from honokiol-treated tumour cells enhanced priming and stimulated T lymphocyte proliferation and type I cytokine production. T lymphocytes stimulated with DCs pulsed with cell lysates of honokiol-treated tumour cells significantly increased specific killing of human cholangiocarcinoma cells compared to those associated with DCs pulsed with cell lysates of untreated cholangiocarcinoma cells. CONCLUSION The present findings suggested that honokiol was able to enhance the immunogenicity of cholangiocarcinoma cells associated with increased effectiveness of DC-based vaccine formulation. Treatment of tumour cells with honokiol offers a promising approach as an ex vivo DC-based anticancer vaccine.

Determination of Magnolol and Honokiol by Non-aqueous Capillary Electrophoresis

Two active principles in traditional Chinese medicine Magnolia officinalis, magnolol and honokiol, were successfully separated by means of nonaqueous capillary electrophoresis. The effect of the composition of a nonaqueous buffer on column efficiency and resolution, and the effect of acid additives on peak shapes were researched. The separation was conducted with a running buffer in a mixture of methanol/aeetonitrile/formamide （ volume ratio ： 1 ： 2 ： 2 ）, in which the concentrations of Tris, acetic acid, and water were 60 retool/L, 0. 04 mmol/L and 5% （ volume fration）, respectively, and the pH^＊ （apperent pH） of the running buffer was 8.96. Magnolol and honokiol were separated on baseline within 20 min. The relative standard deviation of the analytes＇ concentrations in the sample is 1.32% for magnolol and 1.60% for honokiol, and the recoveries of the spiked sample are 98.4% for magnolol and 98. 0% for honokiol, respectively.

Simultaneous Determination of Magnolol and Honokiol rby Synchronous Fluorescence Spectroscopy

A simple sensitive and quick assay for simultaneously determining magnolol （MOL） and honokiol （HOL） has been described based on their natural fluorescence. This method is based on the fact that synchronous fluorometry could resolve the overlapping of fluorescence spectra, which was aroused by their similar molecular structures. In this work, the synchronous spectrum, maintaining a constant difference of Aλ =10 nm between the emission and excitation wavelengths, has been selected for the determination of HOL and MOL. Under the optimum conditions, the fluorescence intensity is proportional to the concentration of MOL and HOL in solution over the range 0.075-0.7 μg/mL and 0.05-0.9 μg/mL with the detection limit of 0.029 μg/mL and 0.019 μg/mL, respectively. The method was applied to the simultaneous determination of MOL and HOL in pharmaceutical dosage with satisfactory results.

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(H_(2)O_(2)),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 H_(2)O_(2)were investigated.Western blotting analysis was used to analyze the expression levels of specific proteins.Results:Honokiol suppressed H_(2)O_(2)-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 H_(2)O_(2)-stimulated HaCaT cells.In addition,honokiol attenuated H_(2)O_(2)-induced reduction of adenosine triphosphate content,and activation of AMP-activated protein kinase(AMPK)was markedly promoted by honokiol in H_(2)O_(2)-stimulated cells.Importantly,the anti-apoptosis and anti-proliferative activity of honokiol against H_(2)O_(2)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.

Synthesis, characterization and in vivo evaluation of honokiol bisphosphate prodrugs protects against rats’ brain ischemia-reperfusion injury

Honokiol(HK)usage is greatly restricted by its poor aqueous solubility and limited oral bioavailability.We synthesized and characterized a novel phosphate prodrug of honokiol(HKP)for in vitro and in vivo use.HKP greatly enhanced the aqueous solubility of HK(127.54±15.53 mg/ml)and the stability in buffer solution was sufficient for intravenous administration.The enzymatic hydrolysis of HKP to HK was extremely rapid in vitro(T 1/2=8.9±2.11 s).Pharmacokinetics studies demonstrated that after intravenous administration of HKP(32 mg/kg),HKP was converted rapidly to HK with a time to reach the maximum plasma concentration of^5 min.The prodrug HKP achieved an improved T 1/2(7.97±1.30 h)and terminal volume of distribution(26.02±6.04 ml/kg)compared with direct injection of the equimolar parent drug(0.66±0.01 h)and(2.90±0.342 ml/kg),respectively.Furthermore,oral administration of HKP showed rapid and improved absorption compared with the parent drug.HKP was confirmed to maintain the bioactivity of the parent drug for ameliorating ischemia-reperfusion injury by decreasing brain infarction and improving neurologic function.Taken together,HKP is a potentially useful aqueous-soluble prodrug with improved pharmacokinetic properties which may merit further development as a potential drug candidate.

Effects of magnolol and honokiol derived from traditional Chinese herbal remedies on gastrointestinal movement

AIM: To study the effects of magnolol and honokiol on isolated smooth muscle of gastrointestinal tract and their relationship with Ca2+, and on the gastric emptying and the intestinal propulsive activity in mice.METHODS: Routine experimental methods using isolated gastric fundus strips of rats and isolated ileum segments of guinea pigs were adopted to measure the smooth muscle tension. The effects of magnolol 10-3, 10-4, 10-5 mol/L, and honokiol 10-4, 10-5, 10-6 mol/L on the contractility of gastric fundus strips of rats and ileum of guinea pigs induced by acetylcholine (Ach) and 5-hydroxytryptamine (5-HT)was assessed respectively. The method using nuclein and pigment methylene blue was adopted to measure the gastric retention rate of nuclein and the intestinal propulsive ratio of a nutritional semi-solid meal for assessing the effect of magnolol and honokiol (0.5, 2, 20 mg/kg) on gastric emptying and intestinal propulsion.RESULTS: Magnolol and honokiol significantly inhibited the contractility of isolated gastric fundus strips of rats treated with Ach or 5-HT and isolated ileum guinea pigs treated with Ach or CaCl2, and both of them behaved as non-competitive muscarinic antagonists. Magnolol and honokiol inhibited the contraction induced by Ach in Ca2+-free medium and extracellular Ca2+-dependent contraction induced by Ach. Each group of magnolol and honokiol experiments significantly decreased the residual rate of nuclein in the stomach and increased the intestinal propulsive ratio in mice.CONCLUSION: The inhibitory effect of magnolol and honokiol on contractility of the smooth muscles of isolated gastric fundus strips of rats and isolated ileum of guinea pigs is associated with a calcium-antagonistic effect.Magnolol and honokiol can improve the gastric emptying of a semi-solid meal and intestinal propulsive activity in mice.

Honokiol Prevents Intestinal Barrier Dysfunction in Mice with Severe Acute Pancreatitis and Inhibits JAK/STAT1 Pathway and Acetylation of HMGB1

Objective To investigate the effect of honokiol(HON)and the role of high-mobility group protein B1(HMGB1)on the pathogenesis of severe acute pancreatitis(SAP).Methods Thirty mice were numbered according to weight,and randomly divided into 5 groups using a random number table,including control,SAP,SAP and normal saline(SAP+NS),SAP and ethyl pyruvate(SAP+EP),or SAP+HON groups,6 mice in each group.Samples of pancreas,intestine,and blood were collected 12 h after SAP model induction for examination of pathologic changes,immune function alterations by enzyme linked immunosorbent assay(ELISA),and Western blot.In vitro experiments,macrophages were divided into 5 groups,the control,lipopolysaccharide(LPS),LPS+DMSO(DMSO),LPS+anti-HMGB1 monoclonal antibody(mAb),and LPS+HON groups.The tight connection level was determined by transmission electron microscopy and fluorescein isothiocyanate-labeled.The location and acetylation of HMGB1 were measured by Western blot.Finally,pyridone 6 and silencing signal transducer and activator of the transcription 1(siSTAT1)combined with honokiol were added to determine whether the Janus kinase(JAK)/STAT1 participated in the regulation of honokiol on HMGB1.The protein expression levels of HMGB1,JAK,and STAT1 were detected using Western blot.Results Mice with SAP had inflammatory injury in the pancreas,bleeding of intestinal tissues,and cells with disrupted histology.Mice in the SAP+HON group had significantly fewer pathological changes.Mice with SAP also had significant increases in the serum levels of amylase,lipase,HMGB1,tumor necrosis factor-α,interleukin-6,diamine oxidase,endotoxin-1,and procalcitonin.Mice in the SAP+HON group did not show these abnormalities(P<0.01).Studies of Caco-2 cells indicated that LPS increased the levels of occludin and claudin-1 as well as tight junction permeability,decreased the levels of junctional adhesion molecule C,and elevated intercellular permeability(P<0.01).HON treatment blocked these effects.Studies of macrophages indicated that LPS led to low nuclear levels of HMGB1,however,HON treatment increased the nuclear level of HMGB1(P<0.01).HON treatment also inhibited the expressions of JAK1,JAK2,and STAT1(P<0.01)and increased the acetylation of HMGB1(P<0.05).Conclusion HON prevented intestinal barrier dysfunction in SAP by inhibiting HMGB1 acetylation and JAK/STAT1 pathway.

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 mitochondrial fission and cell apoptosis by activating Sirt3 in intracerebral hemorrhage

Background:Sirtuin-3(Sirt3)has been documented to protect against mitochondrial dysfunction and apoptosis.Honokiol(HKL)is a Sirt3 pharmacological activator with reported neuroprotective effects in multiple neurological disorders.The present study aimed to explore the neuroprotective effects of HKL and the role of Sirt3 following intracerebral hemorrhage(ICH).Methods:An in vivo ICH model in rats was established by injecting autologous blood into the right basal ganglia.PC12 cells were stimulated with hemin.For the in vivo investigation,the modified Neurological Severity Scores and the Morris water maze test were performed to assess neurological deficits.Hematoxylin-Eosin and Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining were employed to evaluate the histopathology and apoptosis.Immunohistochemical staining was used to investigate the expression of Sirt3.Adenosine triphosphate(ATP)levels were quantified to assess mitochondrial dysfunction.Cell counting kit-8,lactate dehydrogenase assay,and flow cytometry were used to analyze cell vitality and apoptosis in vitro.Immunofluorescence staining was performed to observe mitochondrial morphology and dynamin-related protein 1(Drp1)localization to mitochondria.Western blot was applied to quantify the expression of Sirt3,Bax,Bcl-2,cleaved-caspase-3,Drp1,phosphorylation of Drp1 at serine-616,and phosphorylation of Drp1 at serine-637 in vivo and in vitro.Results:HKL treatment alleviated neurological deficits,attenuated the histopathological damage and cell apoptosis,and restored the decreased ATP levels in ICH rats.HKL improved cell survival rate,reduced cell apoptosis,and inhibited mitochondrial fission in PC12 cells.Moreover,both in vivo and in vitro models showed increased phosphorylation of Drp1 at Ser616,and reduced phosphorylation of Drp1 at Ser637.Meanwhile,immunofluorescence co-localization analysis revealed that hemin increased the overlap of Drp1 and mitochondria in PC12 cells.The phosphorylation and mitochondrial translocation of Drp1 were effectively reversed by HKL treatment.Importantly,the selective Sirt3 inhibitor 3-(1H-1,2,3-triazol-4-yl)pyridine suppressed these effects.Conclusion:Our findings demonstrated that HKL ameliorated ICH-induced apoptosis and mitochondrial fission by Sirt3,suggesting that HKL has immense prospects for the treatment of ICH.

Honokiol attenuates acetaminophen-induced acute liver injury by inhibiting hepatic CYP1A2 activity and improving liver mitochondrial dysfunction

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.

和厚朴酚通过PI3K/AKT信号通路调节线粒体凋亡对小鼠心肌缺血/再灌注损伤的影响及机制研究

目的探讨和厚朴酚(HK)对心肌缺血/再灌注损伤(MIRI)小鼠心脏的保护作用及其机制。方法2022年6月—2023年7月于陕西中医药大学基础医学院中医诊断实验中心进行实验。78只雄性C57BL/6J小鼠随机分为假手术(Sham)组、MIRI组和MIRI+HK组,每组26只。术前对MIRI+HK组小鼠给予HK(0.2 mg·kg-1·d-1)腹腔注射,Sham组和MIRI组给予等量溶剂,连续14 d。造模术中Sham组仅穿线不结扎,其余2组均结扎左前降支30 min后,解开缝线再灌注2 h,构建MIRI模型,再灌注结束后立即取血,剥离心脏。比较各组血清中肌酸激酶同工酶(CK-MB)和心肌肌钙蛋白T(cTnT)水平;超声心动图检测心功能;2,3,5-三苯基氯化四氮唑染色法计算心肌梗死面积;原位末端标记技术检测细胞凋亡水平;电镜观察线粒体结构损伤情况;Western-blot检测线粒体凋亡和磷酸肌醇-3激酶/蛋白激酶B(PI3K/AKT)通路相关蛋白表达。结果与Sham组比较,MIRI组小鼠血清CK-MB、cTnT和左心室收缩末期内径(LVESD)水平及心肌梗死面积、细胞凋亡率升高,左心室射血分数(LVEF)和左心室短轴缩短率(LVFS)降低(P<0.001),心肌细胞线粒体大多肿胀,内外双层膜模糊,嵴畸形且减少或缺失,线粒体凋亡相关蛋白细胞色素(Cyto)-C、Bcl-2相关X蛋白(Bax)和活化半胱氨酸蛋白酶(Cleaved Caspase-9)表达升高,B淋巴细胞瘤2基因(Bcl-2)、p-AKT/AKT和p-PI3K/PI3K表达降低(P<0.001);与MIRI组比较,MIRI+HK组小鼠血清CK-MB、cTnT和LVESD水平及心肌梗死面积、细胞凋亡率降低,LVEF和LVFS增加(P<0.001),心肌细胞线粒体超微结构有所好转,结构较为完整,Cyto-C、Bax和Cleaved Caspase-9蛋白表达降低,Bcl-2、p-AKT/AKT和p-PI3K/PI3K表达升高(P<0.001)。结论HK可通过激活PI3K/AKT信号通路抑制MIRI诱导的心肌细胞线粒体凋亡,从而发挥心肌保护作用。

和厚朴酚的生物学功能及其饲用化前景的研究

和厚朴酚是一种天然的中药提取物,具有抗菌、抗肿瘤、抗应激和抗炎等生物学功能,是中药厚朴发挥药理作用的主要成分。在动物生产中的应用表明其具有改善生产性能、增强免疫力和抗镉中毒等作用。因此,和厚朴酚作为一种新型的饲粮添加剂,具有广泛的应用前景。本文综述了和厚朴酚在机体内的吸收与代谢、生物学功能及其在畜禽生产中的应用,旨在为和厚朴酚在畜禽生产中的高效应用提供参考。

和厚朴酚调控SIRT3抑制慢性缺氧诱导的小胶质细胞极化

目的:探讨和厚朴酚对慢性缺氧条件下小胶质细胞极化的影响及机制。方法:使用ELISA法检测炎症因子,探索氯化钴诱导小胶质细胞系BV2细胞慢性缺氧(48 h)以及和厚朴酚处理的最佳浓度。BV2细胞分为对照、慢性缺氧、慢性缺氧+和厚朴酚、慢性缺氧+和厚朴酚+3-TYP(沉默调节蛋白3,SIRT3抑制剂)4组,ELISA检测上清TNFα及IL-1β蛋白浓度,qPCR检测细胞M1和M2极化标志物表达,生化法检测各组细胞活性氧水平,Western Blot法检测SIRT3和炎症上游分子NLRP3和caspase1蛋白水平。结果:慢性氯化钴刺激BV2细胞最佳浓度为100μmol/L,刺激后其炎症因子TNFα及IL-1β释放较对照组明显增加(P<0.05);与对照组相比,慢性缺氧组细胞SIRT3蛋白表达下调,ROS水平、NLRP3和caspase1蛋白水平,M1极化标志物CD86、iNOS的mRNA水平和CD16/32比值上调。和厚朴酚(10μmol/L)能显著上调慢性缺氧细胞SIRT3蛋白和M2标志物Arg-1及CD206的mRNA水平(P<0.05),下调其ROS、NLRP3/caspase1蛋白和M1标志物转录水平(P<0.05),且这种抗氧化应激和抗炎作用能够被SIRT3抑制剂所逆转。结论:和厚朴酚可抑制慢性缺氧诱导的小胶质细胞M1极化和炎症通路激活,其抗炎作用为SIRT3依赖性。

Inhibition of Herpes Simplex Virus-1 Replication by Natural Compound Honokiol

Honokiol is a pleiotropic natural compound isolated from Magnolia and has multiple biological and clinically relevant effects,including anticancer and antimicrobial function.However,the antiviral activity of honokiol has not yet been well studied.Here we showed that honokiol had no effect on herpes simplex virus-1(HSV-1)entry,but inhibited HSV-1 viral DNA replication,gene expression and the production of new progeny viruses.The combination of honokiol and clinical drug acyclovir augmented inhibition of HSV-1 infection.Our results illustrate that honokiol could be a potential new candidate for clinical consideration in the treatment of HSV-1 infection alone or combination with other therapeutics.

和厚朴酚抑制人脂肪间充质干细胞增殖

目的 研究厚朴酚对人脂肪间充质干细胞(hADSCs)增殖和凋亡的影响,以此细胞模型初探该药物对肿瘤微环境的影响。方法 用不同浓度的和厚朴酚处理hADSCs,分别采用MTS法和台盼蓝染色法检测细胞的增殖能力,annexin V/PI双染法检测细胞凋亡的改变,qPCR和Western blot检测细胞增殖、凋亡相关基因的mRNA和蛋白质表达,Western blot检测MEK-ERK1/2信号通路中总MEK、磷酸化MEK、总ERK和磷酸化ERK蛋白的表达水平。结果 随着浓度增加,和厚朴酚抑制hADSCs增殖、促进其凋亡的作用显著增强。增殖相关基因CCND1、MKI67和PCNA表达下调,促凋亡相关基因BAX和TP53表达上调,抗凋亡基因BCL2表达下调。和厚朴酚呈浓度依赖性抑制MEK和ERK1/2的磷酸化。结论 和厚朴酚抑制hADSCs增殖,促进其凋亡,作用机制可能与抑制MEK-ERK1/2通路活性有关。

Antidiabetic and Anti-oxidative Effects of Honokiol on Diabetic Rats Induced by High-fat Diet and Streptozotocin

Objective To study the antidiabetic and anti-oxidative effects of honokiol （Hon） in Magnolia officinalis and its underlying molecular mechanism in diabetic rats induced by high-fat diet （HFD） and streptozotocin （STZ）. Methods After ig administration with Hon [25, 50, and 100 mg/（kg.d）] to diabetic rats for consecutive 10 weeks, the levels of blood glucose （BG）, oral glucose tolerance （OGT）, blood lipids including total cholesterol （TC）, triglyceride （TG）, high density lipoprotein-cholesterol （HDL-C）, and low density lipoprotein-cholesterol （LDL-C）, hepatic oxidative stress including the activities of superoxide dismutase （SOD）, catalase （CAT）, glutathione peroxidase （GSH-Px）, methane dicarboxylic aldehyde （MDA）, and cytochrome P4502E1 （CYP2E1） in diabetic rats were measured. Results Compared to the diabetic control rats, ig administration of Hon resulted in significant decrease in BC, TC, TG, and LDL-C levels in serum, as well as hepatic CYP2E] activity and MDA content in diabetic rats, whereas the level of OGT and activities of hepatic CAT, SOD, and CSH-Px in diabetic rats were significantly increased. Conclusion Hon could alleviate hyperglycemia, hyperlipemia, hepatic oxidative damage, and insulin resistance in diabetic rats by inhibiting hepatic CYP2E1 activity.

Honokiol: A naturally occurring lignan with pleiotropic bioactivities

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.