Exploring the mechanisms of magnolol in the treatment of periodontitis by integrating network pharmacology and molecular docking

Background:Magnolol,a bioactive extract of the Chinese herb Magnolia officinalis has a protective effect against periodontitis.This study is aimed to explore the mechanisms involved in the functioning of magnolol against periodontitis and provide a basis for further research.Methods:Network pharmacology analysis was performed based on the identification of related targets from public databases.The Protein-protein interaction(PPI)network was constructed to visualize the significance between the targets of magnolol and periodontitis.Subsequently,Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed to predict the functions and the signal regulatory pathways involved in the action of magnolol against periodontitis.The“functiontarget-pathway”networks were constructed to analyze the core targets and pathways of magnolol against periodontitis.Molecular docking was used to verify the interaction of magnolol and core targets.Results:A total of 58 active targets of magnolol and 644 periodontitis-related targets were collected from public databases.A total of 25 targets of magnolol against periodontitis were identified based on the Venn diagram.GO analysis showed that magnolol has a role in the response to oxidative stress,nicotine,and lipopolysaccharide.KEGG enrichment analysis indicated that the mechanism of magnolol against periodontitis was mainly related to the tumor necrosis factor(TNF),phosphoinositide 3-kinase(PI3K/Akt),and mitogen-activated protein kinase(MAPK)signaling pathways.Combined with PPI network and molecular docking results,the core targets of magnolol against periodontitis included AKT1,MAPK8,MAPK14,TNF,and TP53.Conclusion:To summarize,the anti-periodontitis mechanisms of magnolol are potentially through regulating the TNF,PI3K/Akt,and MAPK signaling pathways.

Determination of magnolol and honokiol in traditional Chinese medicine Magnolia officinalis and its preparations by liquid-phase microextraction-back extraction combined with high performance liquid chromatography

Liquid-phase microextraction with back extraction （LPME-BE） combined with high performance liquid chromatography （HPLC） was investigated for the extraction and determination of magnolol and honokiol in Magnolia officinalis, a traditional Chinese medicine （TCM）, and its pharmaceutical preparations, Huo Xiang Zheng Qi peroral liquid and Xiang Sha Yang Wei pellet. Organic solvent, donor and acceptor phases, stirring rate and extraction limes were all factors which can influence the efficiency of extraction and were all optimized during the course of this work. Linear calibration curves were obtained in concentration ranges of 1,56-156 μg/mL for magnolol and 1.10-110 μg/mL for honokiol. Detection limits （S/N = 3） were 0.10 and 0.07 μg/mL, respectively. The relative recoveries were both in the range of 98.3% - 105.1% and RSD was lower than 2.5% .

Magnolol attenuates sepsis-induced gastrointestinal dysmotility in rats by modulating inflammatory mediators

AIM： To investigate the protective effects of magnolol on sepsis-induced inflammation and intestinal dysmotility. METHODS： Sepsis was induced by a single intraperitoneal injection of lipopolysaccharide （LPS）. Male Wistar rats were randomly assigned to one of three treatment groups： magnolol prior to LPS injection （LPS/Mag group）; vehicle prior to LPS injection （LPS/Veh group）; vehicle prior to injection of saline （Control/Veh）. Intestinal transit and circular muscle mechanical activity were assessed 12 h after LPS injection. Tumor necrosis factor-α （TNF-α）, interleukin-10 （IL-10）, monocyte chemoattractant protein-1 （MCP-1） and inducible nitric oxide synthase （iNOS） mRNA in rat ileum were studied by RT-PCR 2 h after LPS injection. Nuclear factor-κB （NF-κB） activity in the intestine was also investigated at this time using electrophoretic mobility shift assay. In addition, antioxidant activity was determined by measuring malondialdehyde （MDA） concentration and superoxide dismutase （SOD） activity in the intestine 2 h after LPS iniection.RESULTS： Magnolol significantly increased intestinal transit and circular muscle mechanical activity in LPS- treated animals. TNF-α, MCP-1 and iNOS mRNA expression in the small intestine were significantly reduced after magnolol treatment in LPS-induced septic animals, compared with untreated septic animals. Additionally,magnolol significantly increased IL-10 mRNA expression in septic rat ileum. Magnolol also significantly suppressed NF-κB activity in septic rat intestine. In addition, magnolol significantly decreased MDA concentration and increased SOD activity in rat ileum. CONCLUSION： Magnolol prevents sepsis-induced suppression of intestinal motility in rats. The potential mechanism of this benefit of magnolol appears to be modulation of self-amplified inflammatory events and block of oxidative stress in the intestine.

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.

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 Ca^2＋, 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 Ca^2＋-free medium and extracellular Ca^2＋-dependent contraction induced by Ach, Each group of magnolol and honokiol experiments significantly decreased the residual rate of nudein 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.

Magnolol protects against acute gastrointestinal injury in sepsis by down-regulating regulated on activation,normal T-cell expressed and secreted

BACKGROUND Sepsis is a major medical challenge.Magnolol is an active constituent of Houpu that improves tissue function and exerts strong anti-endotoxin and anti-inflammatory effects,but the mechanism by which it reduces intestinal inflammation in sepsis is yet unclear.AIM To assess the protective effect of magnolol on intestinal mucosal epithelial cells in sepsis and elucidate the underlying mechanisms.METHODS Enzyme-linked immunosorbent assay was used to measure tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,and regulated on activation,normal T-cell expressed and secreted(RANTES)levels in serum and ileal tissue in animal studies.The histopathological changes of the ileal mucosa in different groups were observed under a microscope.Cell Counting Kit-8 and cell permeability assays were used to determine the concentration of drug-containing serum that did not affect the activity of Caco2 cells but inhibited lipopolysaccharide(LPS)-induced decrease in permeability.Immunofluorescence and Western blot assays were used to detect the levels of RANTES,inhibitor of nuclear factor kappa-B kinaseβ(IKKβ),phosphorylated IKKβ(p-IKKβ),inhibitor of nuclear factor kappa-B kinaseα(IκBα),p65,and p-p65 proteins in different groups in vitro.RESULTS In rats treated with LPS by intravenous tail injection in the presence or absence of magnolol,magnolol inhibited the expression of proinflammatory cytokines,IL-1β,IL-6,and TNF-αin a dose-dependent manner.In addition,magnolol suppressed the production of RANTES in LPS-stimulated sepsis rats.Moreover,in vitro studies suggested that magnolol inhibited the increase of p65 nucleation,thereby markedly downregulating the production of the phosphorylated form of IKKβin LPS-treated Caco2 cells.Specifically,magnolol inhibited the translocation of the transcription factor nuclear factor-kappa B(NF-κB)from the cytosol into the nucleus and down-regulated the expression level of the chemokine RANTES in LPS-stimulated Caco2 cells.CONCLUSION Magnolol down-regulates RANTES levels by inhibiting the LPS/NF-κB signaling pathways,thereby suppressing IL-1β,IL-6,and TNF-αexpression to alleviate the mucosal barrier dysfunction in sepsis.

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.

Magnolol attenuates right ventricular hypertrophy and fibrosis in hypoxia-induced pulmonary arterial hypertensive rats through inhibition of the JAK2/STAT3 signaling pathway

OBJECTIVE Right ventricular(RV)remodeling is one of the essential pathological features in pulmonary arterial hypertension(PAH).RV hypertrophy or fibrosis are the leading causes of RV remodeling.Magnolol is a compound isolated from Magnolia officinalis.It possesses multiple pharmacological activities,such as anti-oxidation and anti-inflammation.This study aims to evaluate the effects and underlying mechanisms of magnolol on RV remodeling in hypoxia-induced PAH.METHODS①Male SD rats(220 g)were randomly divided into 5 groups(n=10):the normoxia group,the hypoxia group,the hypoxia plus Magnolol(10 and 20 mg·kg^(-1)·d-1)group,and the vehicle group.Rats in the normoxia group were kept in a normoxia environment for 4 weeks,while rats in the hypoxia group were kept in a hypoxic chamber(10%O2).The rats in the hypoxia plus magnolol groups were administered with magnolol at 10 or 20 mg·kg^(-1)(ip)once a day for 4 weeks.At the end of 4 weeks,the heart function was assessed by Doppler echocardiography,and then the rats were anesthetized with sodium pentobarbital(30 mg·kg^(-1),ip).The RVSP was measured by the right heart catheterization method.The heart tissues were collected and dissected to calculate the index of RV remodeling(RV/LV+IVS,RV/tibial length,or RV/body weight).Part of the RV samples was fixed with 4%paraformaldehyde for morphological analysis,while other samples were frozen at-80℃for molecular studies(measurements of ANP,BNP,α-SMA,and collagenⅠ/ⅢmRNA expression as well as p-JAK2/JAK2 and p-STAT3/STAT3 protein levels).②To evaluate the effect of magnolol on hypoxia-induced myocardial hypertrophy and fibrosis,H9c2 or cardiac fibroblasts were divided into 7 groups:the control group,cells were cultured under normal conditions;the hypoxia group,cells were cultured under hypoxic condition(3%O2);the hypoxia plus magnolol 10 mg·kg^(-1) group,magnolol10μmol·L^(-1) was added to the culture medium before the hypoxia treatment;the hypoxia plus magnolol 30 mg·kg^(-1) group,magnolol 20μmol·L^(-1) was added to the culture medium before the hypoxia treatment;the hypoxia plus TG-101348 group,TG-101348(a specific inhibitor of JAK2)1μmol·L^(-1) was added to the culture medium before the hypoxia treatment;the hypoxia plus JSI-124 group,JSI-124(a specific inhibitor of JAK2)1μmol·L^(-1) was added to the culture medium before the hypoxia treatment;and the hypoxia plus vehicle group,an equal volume of vehicle(DMSO)was added to the culture medium before the hypoxia treatment.At the end of the experiments,the cells were collected for morphological and molecular analysis.RESULTS In vivo,male Sprang-Daley rats were exposed to 10%O2 for 4 weeks to establish an RV remodeling model,which showed hypertrophic and fibrotic features(increases of RV remodeling index,cellular size,hypertrophic and fibrotic marker expression),accompanied by an elevation in phosphorylation levels of JAK2 and STAT3;these changes were attenuated by treating rats with magnolol.In vitro,the cultured H9c2 cells or cardiac fibroblasts were exposed to 3%O2 for 48 h to induce hypertrophy or fibrosis,which showed hypertrophic(increases in cellular size as well as the expression of ANP and BNP)or fibrotic features(increases in the expression of collagenⅠ,collagenⅢandα-SMA).Administration of magnolol and TG-101348 or JSI-124 (JAK2 selective inhibitors) could prevent the process of myocardial hypertrophy and fibrosis, accompanied by the decrease in the phosphorylation level of JAK2 and STAT3. CONCLUSION Magnolol can attenuate RV hypertrophy and fibrosis in hypoxia-induced PAH rats through a mechanism involving inhibition of the JAK2/STAT3 signaling pathway.

Ionic liquids functionalized β-cyclodextrin polymer for separation/analysis of magnolol

Ionic liquids functionalized β-cyclodextrin polymer, a mono-6-deoxy-6-（1,2-dimethylimida- zolium）-β-cyclodextrin iodide polymer （ILs-β-CDCP）, was synthesized as a solid-phase adsorbent coupled with high-performance liquid chromatography for separating or analyzing magnolol in drug samples. The results showed that magnolol was adsorbed rapidly on ILs-D-CDCP and eluted with methanol. Under the optimum conditions, preconcentration factor of the proposed method was 12. The linear range, limit of detection （LOD）, correlation coefficient （R） and relative standard deviation （RSD） were found to be 0.02-8.00 μg/mL, 1.9 ng/mL, 0.9992 and 2.76% （n=3, c=2.00 pg/mL）, respectively. The interaction between 1Ls-]）-CDCP and magnolol was studied through the inclusion constant, FTIR and TGA analysis. This proposed method has been successfully applied to the determination of magnolol in real samples.

Simultaneous Determination of Magnolol and Honokiol by 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.

Preliminary study on the anti-inflammatory effect of magnolol on LPS-induced mice

Objective:Investigating the anti-inflammatory effects of magnolol on lipopolysaccharide (LPS)-induced inflammation model mouse and its effect on NF-κB pathway.Methods:Mice were randomly divided into 5 group: normal control group, inflammatory model group, dexamethasone positive drug group and, magnolol group. After administrating for 7 d, LPS was intraperitoneally injected to induce inflammatory on the 8th day, and blood was taken 3 h later and the thymus and spleen were weighed. The levels of serum TNF-α, IL-22 and IL-17 were detected by ELISA method. The expression levels of TNF-α, NF-κB p65 and IL-17 in thymus were detected by immunohistochemistry.Results: Magnolol could evidently reduce the levels of serum TNF-α, NF-κB p65 and IL-17 and IL-22. Immunohistochemistry result showed that magnolol could down-regulate the expression of IL-17, TNF-α and p65. Conclusion: Magnolol can prevent and treat LPS-induced inflammation. Its anti-inflammatory effect is connected with the down-regulation of TNF-α and NF-κB p65 protein levels by IL-17 inflammatory pathway. This experiment provides a theoretical basis for magnolol in anti-inflammatory.

Magnolol and 5-fluorouracil synergy inhibition of metastasis of cervical cancer cells by targeting PI3K/AKT/mTOR and EMT pathways

Objective: This study is designed to investigate the mode of action of the synergistic effect of 5-fluorouracil(5-FU) and magnolol against cervical cancer.Methods: Network pharmacological approach was applied to predict the molecular mechanism of 5-FU combined with magnolol against cervical cancer. CCK-8 assay, colony formation assay, immunofluorescence staining, adhesion assay, wound healing mobility assay, cell migration and invasion assay and Western blot analysis were conducted to validate the results of in silico study.Results: Phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR) signaling pathway was identified as the key pathway in silico study. The experimental results showed that 5-FU combined with magnolol strongly inhibited cervical cancer cell proliferation, induced the morphological change of HeLa cells by down-regulating the expression of a-actinin, tensin-2 and vinculin. Moreover, magnolol enhanced inhibitory effect of 5-FU on the cell adhesion, migration and invasion. The phosphorylation of AKT and PI3K and the expression of mTOR were strongly inhibited by the combination of 5-FU and magnolol. Moreover, the expression of E-cadherin and β-catenin was upregulated and the expression of Snail, Slug and vimentin was down-regulated by the 5-FU together with magnolol.Conclusion: Taken together, this study suggests that 5-FU combined with magnolol exerts a synergistic anti-cervical cancer effect by regulating the PI3K/AKT/mTOR and epithelial-mesenchymal transition(EMT) signaling pathways.

生物质基厚朴酚和糠醛对酚醛树脂的改性研究

为了减少化石能源的使用,选取生物质基厚朴酚和糠醛取代部分苯酚和甲醛,制备了改性酚醛树脂。通过傅里叶变换红外光谱仪,核磁共振波谱仪对改性酚醛树脂进行表征,考察了改性酚醛树脂的基础性能及固化后的力学性能。结果表明:厚朴酚和糠醛改变了酚醛树脂的分子结构,改性酚醛树脂的黏度和游离酚含量明显降低;经过固化后的改性酚醛树脂拉伸强度、冲击强度、弯曲强度分别提高18.6%、20.4%和24.9%。

厚朴酚环氧树脂涂层的制备及综合防护性能研究

采用联苯结构的生物质原料厚朴酚合成生物基环氧树脂,以柔性聚醚胺固化剂(D230、D400、D2000及T403)进行固化制备生物基环氧抗冲蚀涂层,探究聚醚胺分子量与官能度对厚朴酚环氧树脂力学性能、防腐性能以及抗冲蚀防护性能的影响,阐述厚朴酚环氧涂层抗冲蚀磨损机理。结果表明:D230固化的厚朴酚环氧涂层在3.5%(质量分数)NaCl溶液中浸泡15 d后仍具有较高的阻抗值(1.25×10^(11)Ω·cm^(2));以T403固化可获得高拉伸强度(26.59 MPa)、低摩擦系数(0.373)和磨损率[0.0183 mm^(3)/(N·m)]涂层;D2000固化的厚朴酚环氧涂层在抗冲蚀测试中表现出较低质量损失(244.74 mg)与体积损失(157 mm^(3))。在固/液/气三相流冲蚀作用下,D2000较长的柔性链段,赋予厚朴酚环氧涂层更好的弹性和柔韧性,降低了冲蚀介质的冲击作用,从而表现出优异的抗冲蚀性能。

厚朴酚对动物肠道黏膜屏障功能的影响及其调控机制

肠道黏膜屏障作为机体与外部环境进行互作的重要场所,可防止肠道内病原微生物和有毒有害物质侵入,其完整性对动物肠道健康至关重要。厚朴酚是我国传统中药厚朴中提取的活性成分之一,具有抗癌、抗菌、抗炎和抗氧化等功能。研究发现,厚朴酚可通过调控肠道菌群的组成、促进短链脂肪酸和碱性磷酸酶的生成、抑制肠上皮细胞的凋亡、上调肠上皮细胞紧密连接蛋白的表达、调节细胞因子的分泌和增强抗氧化酶的活性等途径,进而维护肠道黏膜屏障的完整性。本文主要就厚朴酚对动物肠道黏膜屏障功能的影响及其调控机制进行综述,为厚朴酚在畜禽生产中的合理应用提供参考。

Magnolol additive as a replacer of antibiotic enhances the growth performance of Linwu ducks

Magnolol rich in Magnolia officinalis is a bioactive polyphenolic compound. The aim of this study was to examine the effects of magnolol additive(MA) on growth performance, expression levels of antioxidantrelated genes, and intestinal mucosal morphology of Linwu ducks aged from 49 to 70 days, comparing with that of an antibiotic additive(colistin sulfate [CS]). A total of 275,49-day-old ducks were assigned to5 groups with 5 cages of 11 ducks each and fed diets supplemented with 0,100, 200 and 300 mg of MA/kg and 300 mg of CS/kg for 3 weeks, respectively. The results showed that the average daily body weight gain(ADG) was increased significantly in MA-fed groups(200 and 300 mg/kg), compared with the basal diet(BD) group(P < 0.05). The mRNA levels of superoxide dismutase-1(SOD1), manganese superoxide dismutase-2(MnSOD2) and catalase(CAT) were also increased significantly in MA groups(P < 0.05). In addition, hematoxylin and eosin staining revealed that Linwu ducks fed the diets with MA had more intact intestinal mucosa than those fed the BD and CS diets. In addition, ileal villus height, ileal villus height/crypt depth ratio(V/C) and duodenal V/C were also improved significantly(P < 0.05). Taken together, these data demonstrated that MA is an effective feed additive to enhance the growth performance of the Linwu ducks by improving the antioxidant and intestinal mucosal status, suggesting that MA will be a potential additive to replace antibiotic(CS).

厚朴酚、和厚朴酚对脂多糖诱导小鼠肠道损伤的抗炎作用及机制研究

试验旨在研究厚朴酚、和厚朴酚对脂多糖诱导小鼠的肠道损伤的抗炎作用。选用体重为18~22 g的ICR健康雄性小鼠135只,随机分为9组,每组3个重复,每个重复5只小鼠。各组分别为对照组、模型组、阳性药物组(灌胃100 mg/kg盐酸小檗碱)以及厚朴酚、和厚朴酚的低、中、高剂量组(25、50、100 mg/kg)。造模前小鼠按照试验分组连续灌胃7 d,对照组和模型组灌胃生理盐水。第7 d除对照组外,其余组小鼠均腹腔注射6 mg/kg脂多糖诱导炎症。结果显示,厚朴酚、和厚朴酚能够降低腹泻指数和促炎细胞因子白细胞介素-6的水平,减轻肠炎症状,改善肠道形态,恢复肠道微生物群平衡;并且发现蛋白激酶B(AKT)、B细胞淋巴瘤细胞凋亡抑制因子(BCL-XL)和白细胞介素-18受体1(IL-18R1)是炎症的潜在治疗靶点,厚朴酚通过上调AKT、BCL-XL等基因,激活磷脂酰肌醇3激酶-蛋白激酶(BPI3K-Akt)信号通路发挥抗炎作用。相反,和厚朴酚的抗炎机制涉及下调炎症相关基因,包括IL-18R1和环磷腺苷效应元件结合蛋白(CREB),从而抑制肿瘤坏死因子(TNF)信号通路。研究表明,在缓解治疗肠道损伤方面,推荐厚朴酚、和厚朴酚的适宜添加量为100 mg/kg。

Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury

Objective： Heat stress （HS） is an important environmental stressor that adversely influences livestock during the summer. The aim of this study was to investigate whether magnolol protects against HS-induced intestinal epithelial cell injury. Materials and methods： An intestinal epithelial cell line （IEC-6） was subjected to HS at 42℃, with and without magnolol pretreatment. Cell injury was detected by monitoring lactate dehydrogenase （LDH） release. MTS （3-（4,5-dimethyithiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium） assay was used to as- sess cell proliferation and viability, including identifying effective concentrations of magnolol. Flow cytometry confirmed Gl-phase cell-cycle arrest and its alleviation by magnolol. Active DNA synthesis was measured by incorporation of nucleic acid 5-ethynyl-2＇-deoxyuridine （EdU）. Gl-phase cell-cycle-related gene expression was assessed by real-time reverse transcription polymerase chain reaction （RT-PCR） and levels of Gl-phase-related proteins by Western blotting Results： HS induced IEC-6 cell injury and decreased cell viability, as demonstrated by data from LDH and MTS assays respectively. Based on a number of criteria, IEC-6 cells subjected to HS were arrested in the G1 phase Of the cell cycle. Magnolol pretreatment decreased HS-induced cell injury through relief of this cell-cycle arrest. Conclusions： Magnolol pretreatment attenuates HS-induced injury in IEC-6 cells. Magnolol is potentially promising as a protective strategy for HS in livestock.

厚朴酚对牙周炎大鼠牙周组织中IL-6、VEGF表达的影响

目的观察厚朴酚对牙周炎大鼠牙周病变及牙周组织中白细胞介素-6(IL-6)、血管内皮生长因子(VEGF)表达的影响,探讨厚朴酚治疗牙周炎的价值。方法取3月龄雄性SD大鼠65只,随机选取其中10只作为对照组,其余大鼠采用正畸结扎丝结扎双侧上颌第一磨牙方法建立牙周炎模型。结扎4周后,将造模成功的40只大鼠随机分为4组,每组10只。局部治疗组去除局部刺激因素即结扎丝;厚朴酚治疗组在不去除结扎丝的情况下给予厚朴酚50 mg/(kg·d)灌胃,局部+厚朴酚治疗组在去除结扎丝的同时给予厚朴酚50 mg/(kg·d)灌胃,对照组给予等剂量生理盐水灌胃,各组均干预8周。观察各组大鼠牙体牙周组织一般情况,HE染色观察牙周组织病理形态,免疫组化染色检测牙周组织中IL-6、VEGF阳性表达情况。结果牙周炎组大鼠双侧上颌第一磨牙龈缘红肿;结合上皮离开釉牙骨质界向根方移位形成深牙周袋,周围有大量炎细胞浸润,牙槽嵴顶及固有牙槽骨吸收破坏。局部治疗组双侧上颌第一磨牙龈红肿,探诊出血;牙周袋深,周围有少量炎细胞浸润,牙槽嵴顶及固有牙槽骨破坏严重,牙槽嵴顶可见新生骨形成。厚朴酚治疗组和局部+厚朴酚治疗组双侧上颌第一磨牙龈红肿较轻或无红肿;牙周袋浅,周围有少量炎性细胞浸润,牙槽嵴顶均可见新生骨形成,其中局部+厚朴酚治疗组牙周组织破坏更轻。牙周炎组大鼠牙周组织中IL-6及VEGF阳性表达平均光密度值均明显高于对照组(P均<0.05),厚朴酚治疗组和局部+厚朴酚治疗组大鼠牙周组织中IL-6及VEGF阳性表达平均光密度值均明显低于牙周炎组(P均<0.05),且局部+厚朴酚治疗组均明显低于局部治疗组(P均<0.05)。结论厚朴酚能够显著降低牙周炎大鼠牙周组织中的致炎细胞因子水平,减轻牙周病变,改善牙槽骨的新生状态。

Synergistic effects of autophagy/mitophagy inhibitors and magnolol promote apoptosis and antitumor efficacy

Mitochondria as a signaling platform play crucial roles in deciding cell fate.Many classic anticancer agents are known to trigger cell death through induction of mitochondrial damage.Mitophagy,one selective autophagy,is the key mitochondrial quality control that effectively removes damaged mitochondria.However,the precise roles of mitophagy in tumorigenesis and anticancer agent treatment remain largely unclear.Here,we examined the functional implication of mitophagy in the anticancer properties of magnolol,a natural product isolated from herbal Magnolia officinalis.First,we found that magnolol induces mitochondrial depolarization,causes excessive mitochondrial fragmentation,and increases mitochondrial reactive oxygen species(mtROS).Second,magnolol induces PTEN-induced putative kinase protein 1(PINK1)-Parkin-mediated mitophagy through regulating two positive feedforward amplification loops.Third,magnolol triggers cancer cell death and inhibits neuroblastoma tumor growth via the intrinsic apoptosis pathway.Moreover,magnolol prolongs the survival time of tumor-bearing mice.Finally,inhibition of mitophagy by PINK1/Parkin knockdown or using inhibitors targeting different autophagy/mitophagy stages significantly promotes magnolol-induced cell death and enhances magnolol's anticancer efficacy,both in vitro and in vivo.Altogether,our study demonstrates that magnolol can induce autophagy/mitophagy and apoptosis,whereas blockage of autophagy/mitophagy remarkably enhances the anticancer efficacy of magnolol,suggesting that targeting mitophagy may be a promising strategy to overcome chemoresistance and improve anticancer therapy.