Ginsenoside Rb1 protects dopaminergic neurons from inflammatory injury induced by intranigral lipopolysaccharide injection

详细信息    查看全文 | 推荐本文 |

英文篇名：Ginsenoside Rb1 protects dopaminergic neurons from inflammatory injury induced by intranigral lipopolysaccharide injection 作者：Da-Wei ; Li ; Fa-Zhan ; Zhou ; Xian-Chang ; Sun ; Shu-Chen ; Li ; Jin-Bin ; Yang ; Huan-Huan ; Sun ; Ai-Hua ; Wang 英文作者：Da-Wei Li;Fa-Zhan Zhou;Xian-Chang Sun;Shu-Chen Li;Jin-Bin Yang;Huan-Huan Sun;Ai-Hua Wang;Department of Neurology, Qianfoshan Hospital Affiliated to Shandong University;Department of Neurology, The People's Hospital of Xintai;Department of Cardiovascularology, Taian Central Hospital;Department of Physiology, Taishan Medical University; 英文关键词：nerve regeneration;;neurodegeneration;;Parkinson's disease;;ginsenoside Rb1;;neuroinflammation;;lipopolysaccharide;;dopaminergic neuron;;microglia;;nuclear factor kappa B;;dopamine;;tyrosine hydroxylase;;substantia nigra;;neural regeneration 中文刊名：SJZY 英文刊名：中国神经再生研究(英文版) 机构：Department of Neurology, Qianfoshan Hospital Affiliated to Shandong University;Department of Neurology, The People's Hospital of Xintai;Department of Cardiovascularology, Taian Central Hospital;Department of Physiology, Taishan Medical University; 出版日期：2019-07-10 出版单位：Neural Regeneration Research 年：2019 期：v.14 基金：supported by the Medical and Health Technology Development Plan of Shandong Province of China,No.2011HD009(to AHW);; the Chinese Medicine Science and Technology Development Plan Project of Shandong Province of China,No.2017-163(to AHW);; the Natural Science Foundation of Shandong Province of China,No.ZR2016HP23(to AHW);; the Science and Technology Development Plan Project of Taian City of China,No.2017NS0151(to XCS) 语种：英文; 页：SJZY201910030 页数：9 CN：10 ISSN：11-5422/R 分类号：160-168

摘要

Accumulating studies suggest that neuroinflammation characterized by microglial overactivation plays a pivotal role in the pathogenesis of Parkinson's disease.As such,inhibition of microglial overactivation might be a promising treatment strategy to delay the onset or slow the progression of Parkinson's disease.Ginsenoside Rbl,the most active ingredient of ginseng,reportedly exerts neuroprotective effects by suppressing inflammation in vitro.The present study aimed to evaluate the neuroprotective and anti-inflammatory effects of ginsenoside Rbl in a lipopolysaccharide-induced rat Parkinson's disease model.Rats were divided into four groups.In the control group,sham-operated rats were intraperitoneally administered normal saline for 14 consecutive days.In the ginsenoside Rbl group,ginsenoside Rb1(20 mg/kg)was intraperitoneally injected for 14 consecutive days after sham surgery.In the lipopolysaccharide group,a single dose of lipopolysaccharide was unilaterally microinjected into the rat substantial nigra to establish the Parkinson's disease model.Lipopolysaccharide-injected rats were treated with normal saline for 14 consecutive days.In the ginsenoside Rbl +lipopolysaccharide group,lipopolysaccharide was unilaterally microinjected into the rat substantial nigra.Subsequently,ginsenoside Rbl was intraperitoneally injected for 14 consecutive days.To investigate the therapeutic effects of ginsenoside Rbl,behavioral tests were performed on day 15 after lipopolysaccharide injection.We found that ginsenoside Rbl treatment remarkably reduced apomorphine-induced rotations in lipopolysaccharide-treated rats compared with the lipopolysaccharide group.To investigate the neurotoxicity of lipopolysaccharide and potential protective effect of ginsenoside Rbl,contents of dopamine and its metabolites in the striatum were measured by high-performance liquid chromatography.Compared with the lipopolysaccharide group,ginsenoside Rbl obviously attenuated the lipopolysaccharide-induced depletion of dopamine and its metabolites in the striatum.To further explore the neuroprotective effect of ginsenoside Rbl against lipopolysaccharide-induced neurotoxicity,immunohistochemistry and western blot assay of tyrosine hydroxylase were performed to evaluate dopaminergic neuron degeneration in the substantial nigra par compacta.The results showed that lipopolysaccharide injection caused a large loss of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra and a significant decrease in overall tyrosine hydroxylase expression.However,ginsenoside Rb1 noticeably reversed these changes.To investigate whether the neuroprotective effect of ginsenoside Rbl was associated with inhibition of lipopolysaccharide-induced microglial activation,we examined expression of the microglia marker Iba-1.Our results confirmed that lipopolysaccharide injection induced a significant increase in Iba-1 expression in the substantia nigra;however,ginsenoside Rbl effectively suppressed lipopolysaccharide-induced microglial overactivation.To elucidate the inhibitory mechanism of ginsenoside Rb1,we examined expression levels of inflammatory mediators(tumor necrosis factor-a,interleukin-1β,inducible nitric oxide synthase,and cyclooxygenase 2)and phosphorylation of nuclear factor kappa B signaling-related proteins(IκB,IKK)in the substantia nigra with enzyme-linked immunosorbent and western blot assays.Our results revealed that compared with the control group,phosphorylation and expression of inflammatory mediators IκB and IKK in the substantia nigra of lipopolysaccharide group rats were significantly increased;whereas,ginsenoside Rbl obviously reduced lipopolysaccharide-induced changes on the lesioned side of the substantial nigra par compacta.These findings confirm that ginsenoside Rbl can inhibit inflammation induced by lipopolysaccharide injection into the substantia nigra and protect dopaminergic neurons,which may be related to its inhibition of the nuclear factor kappa B signaling pathway.This study was approved by the Experimental Animal Ethics Committee of Shandong University of China in April 2016(approval No.KYLL-2016-0148).
        Accumulating studies suggest that neuroinflammation characterized by microglial overactivation plays a pivotal role in the pathogenesis of Parkinson's disease.As such,inhibition of microglial overactivation might be a promising treatment strategy to delay the onset or slow the progression of Parkinson's disease.Ginsenoside Rbl,the most active ingredient of ginseng,reportedly exerts neuroprotective effects by suppressing inflammation in vitro.The present study aimed to evaluate the neuroprotective and anti-inflammatory effects of ginsenoside Rbl in a lipopolysaccharide-induced rat Parkinson's disease model.Rats were divided into four groups.In the control group,sham-operated rats were intraperitoneally administered normal saline for 14 consecutive days.In the ginsenoside Rbl group,ginsenoside Rb1(20 mg/kg)was intraperitoneally injected for 14 consecutive days after sham surgery.In the lipopolysaccharide group,a single dose of lipopolysaccharide was unilaterally microinjected into the rat substantial nigra to establish the Parkinson's disease model.Lipopolysaccharide-injected rats were treated with normal saline for 14 consecutive days.In the ginsenoside Rbl +lipopolysaccharide group,lipopolysaccharide was unilaterally microinjected into the rat substantial nigra.Subsequently,ginsenoside Rbl was intraperitoneally injected for 14 consecutive days.To investigate the therapeutic effects of ginsenoside Rbl,behavioral tests were performed on day 15 after lipopolysaccharide injection.We found that ginsenoside Rbl treatment remarkably reduced apomorphine-induced rotations in lipopolysaccharide-treated rats compared with the lipopolysaccharide group.To investigate the neurotoxicity of lipopolysaccharide and potential protective effect of ginsenoside Rbl,contents of dopamine and its metabolites in the striatum were measured by high-performance liquid chromatography.Compared with the lipopolysaccharide group,ginsenoside Rbl obviously attenuated the lipopolysaccharide-induced depletion of dopamine and its metabolites in the striatum.To further explore the neuroprotective effect of ginsenoside Rbl against lipopolysaccharide-induced neurotoxicity,immunohistochemistry and western blot assay of tyrosine hydroxylase were performed to evaluate dopaminergic neuron degeneration in the substantial nigra par compacta.The results showed that lipopolysaccharide injection caused a large loss of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra and a significant decrease in overall tyrosine hydroxylase expression.However,ginsenoside Rb1 noticeably reversed these changes.To investigate whether the neuroprotective effect of ginsenoside Rbl was associated with inhibition of lipopolysaccharide-induced microglial activation,we examined expression of the microglia marker Iba-1.Our results confirmed that lipopolysaccharide injection induced a significant increase in Iba-1 expression in the substantia nigra;however,ginsenoside Rbl effectively suppressed lipopolysaccharide-induced microglial overactivation.To elucidate the inhibitory mechanism of ginsenoside Rb1,we examined expression levels of inflammatory mediators(tumor necrosis factor-a,interleukin-1β,inducible nitric oxide synthase,and cyclooxygenase 2)and phosphorylation of nuclear factor kappa B signaling-related proteins(IκB,IKK)in the substantia nigra with enzyme-linked immunosorbent and western blot assays.Our results revealed that compared with the control group,phosphorylation and expression of inflammatory mediators IκB and IKK in the substantia nigra of lipopolysaccharide group rats were significantly increased;whereas,ginsenoside Rbl obviously reduced lipopolysaccharide-induced changes on the lesioned side of the substantial nigra par compacta.These findings confirm that ginsenoside Rbl can inhibit inflammation induced by lipopolysaccharide injection into the substantia nigra and protect dopaminergic neurons,which may be related to its inhibition of the nuclear factor kappa B signaling pathway.This study was approved by the Experimental Animal Ethics Committee of Shandong University of China in April 2016(approval No.KYLL-2016-0148).

引文

Ahmed T,Raza SH,Maryam A,Setzer WN,Braidy N,Nabavi SF,de Oliveira MR,Nabavi SM(2016)Ginsenoside Rb1 as a neuroprotective agent:a review.Brain Res Bull 125:30-43.
    Alzoubi KH,Mokhemer E,Abuirmeileh AN(2018)Beneficial effect of etazolate on depression-like behavior and,learning,and memory impairment in a model of Parkinson’s disease.Behav Brain Res 350:109-115.
    Ardah MT,Paleologou KE,Lv G,Menon SA,Abul Khair SB,Lu JH,Safieh-Garabedian B,Al-Hayani AA,Eliezer D,Li M,El-Agnaf OM(2015)Ginsenoside Rb1 inhibits fibrillation and toxicity of alpha-synuclein and disaggregates preformed fibrils.Neurobiol Dis 74:89-101.
    Beamer CA,Shepherd DM(2012)Inhibition of TLR ligand-and interferon gamma-induced murine microglial activation by Panax notoginseng.JNeuroimmune Pharmacol 7:465-476.
    Bjorklund A,Dunnett SB(2019)The amphetamine induced rotation test:a re-assessment of its use as a tool to monitor motor impairment and functional recovery in rodent models of Parkinson’s disease.J Parkinsons Dis9:17-29.
    Castano A,Herrera AJ,Cano J,Machado A(1998)Lipopolysaccharide intranigral injection induces inflammatory reaction and damage in nigrostriatal dopaminergic system.J Neurochem 70:1584-1592.
    Chen W,Guo Y,Yang W,Zheng P,Zeng J,Tong W(2015)Protective effect of ginsenoside Rb1 on integrity of blood-brain barrier following cerebral ischemia.Exp Brain Res 233:2823-2831.
    Chen X,Huang T,Zhang J,Song J,Chen L,Zhu Y(2008)Involvement of calpain and p25 of CDK5 pathway in ginsenoside Rb1’s attenuation of beta-amyloid peptide 25-35-induced tau hyperphosphorylation in cortical neurons.Brain Res 1200:99-106.
    Cho J,Park W,Lee S,Ahn W,Lee Y(2004)Ginsenoside-Rb1 from Panax ginseng C.A.Meyer activates estrogen receptor-alpha and-beta,independent of ligand binding.J Clin Endocrinol Metab 89:3510-3515.
    Erny D,Hrabe de Angelis AL,Jaitin D,Wieghofer P,Staszewski O,David E,Keren-Shaul H,Mahlakoiv T,Jakobshagen K,Buch T,Schwierzeck V,Utermohlen O,Chun E,Garrett WS,McCoy KD,Diefenbach A,Staeheli P,Stecher B,Amit I,Prinz M(2015)Host microbiota constantly control maturation and function of microglia in the CNS.Nat Neurosci 18:965-977.
    Fernandez-Moriano C,Gonzalez-Burgos E,Iglesias I,Lozano R,Gomez-Serranillos MP(2017)Evaluation of the adaptogenic potential exerted by ginsenosides Rb1 and Rg1 against oxidative stress-mediated neurotoxicity in an in vitro neuronal model.PLoS One 12:e0182933.
    Gupta N,Shyamasundar S,Patnala R,Karthikeyan A,Arumugam TV,Ling EA,Dheen ST(2018)Recent progress in therapeutic strategies for microglia-mediated neuroinflammation in neuropathologies.Expert Opin Ther Targets 22:765-781.
    Hashimoto R,Yu J,Koizumi H,Ouchi Y,Okabe T(2012)Ginsenoside Rb1 prevents MPP(+)-induced apoptosis in PC12 cells by stimulating estrogen receptors with consequent activation of ERK1/2,akt and inhibition of SAPK/JNK,p38 MAPK.Evid Based Complement Alternat Med2012:693717.
    Herrera AJ,Castano A,Venero JL,Cano J,Machado A(2000)The single intranigral injection of LPS as a new model for studying the selective effects of inflammatory reactions on dopaminergic system.Neurobiol Dis 7:429-447.
    Hirsch EC,Hunot S(2009)Neuroinflammation in Parkinson’s disease:a target for neuroprotection?Lancet Neurol 8:382-397.
    Hritcu L,Ciobica A(2013)Intranigral lipopolysaccharide administration induced behavioral deficits and oxidative stress damage in laboratory rats:relevance for Parkinson’s disease.Behav Brain Res 253:25-31.
    Inoue M,Oishi N,Fukuyama H(2010)A MRI template of rat brain matched to the Paxinos and Watson atlas sixth edition.Neurosci Res68:E444.
    Ke L,Guo W,Xu J,Zhang G,Wang W,Huang W(2014)Ginsenoside Rb1attenuates activated microglia-induced neuronal damage.Neural Regen Res 9:252-259.
    Le W,Wu J,Tang Y(2016)Protective microglia and their regulation in parkinson’s disease.Front Mol Neurosci 9:89.
    Lee JS,Song JH,Sohn NW,Shin JW(2013)Inhibitory effects of ginsenoside Rb1 on neuroinflammation following systemic lipopolysaccharide treatment in mice.Phytother Res 27:1270-1276.
    Lee KW,Jung SY,Choi SM,Yang EJ(2012)Effects of ginsenoside Re on LPS-induced inflammatory mediators in BV2 microglial cells.BMCComplement Altern Med 12:196.
    Lee Y,Lee S,Chang SC,Lee J(2019)Significant roles of neuroinflammation in Parkinson’s disease:therapeutic targets for PD prevention.Arch Pharm Res doi:10.1007/s12272-019-01133-0.
    Liu D,Zhang H,Gu W,Liu Y,Zhang M(2013)Neuroprotective effects of ginsenoside Rb1 on high glucose-induced neurotoxicity in primary cultured rat hippocampal neurons.PLoS One 8:e79399.
    Liu LX,Chen WF,Xie JX,Wong MS(2008)Neuroprotective effects of genistein on dopaminergic neurons in the mice model of Parkinson’s disease.Neurosci Res 60:156-161.
    Liu X,Gu X,Yu M,Zi Y,Yu H,Wang Y,Xie Y,Xiang L(2018)Effects of ginsenoside Rb1 on oxidative stress injury in rat spinal cords by regulating the eNOS/Nrf2/HO-1 signaling pathway.Exp Ther Med 16:1079-1086.
    Machado A,Herrera AJ,Venero JL,Santiago M,de Pablos RM,Villaran RF,Espinosa-Oliva AM,Arguelles S,Sarmiento M,Delgado-Cortes MJ,Maurino R,Cano J(2011)Inflammatory animal model for Parkinson’s disease:the intranigral injection of lps induced the inflammatory process along with the selective degeneration of nigrostriatal dopaminergic neurons.ISRN neurol 2011:476158.
    Miao HH,Zhang Y,Ding GN,Hong FX,Dong P,Tian M(2017)Ginsenoside Rb1 attenuates isoflurane/surgery-induced cognitive dysfunction via inhibiting neuroinflammation and oxidative stress.Biomed Environ Sci30:363-372.
    Miyanishi K,Choudhury ME,Watanabe M,Kubo M,Nomoto M,Yano H,Tanaka J(2019)Behavioral tests predicting striatal dopamine level in a rat hemi-Parkinson’s disease model.Neurochem Int 122:38-46.
    Nolan YM,Sullivan AM,Toulouse A(2013)Parkinson’s disease in the nuclear age of neuroinflammation.Trends Mol Med 19:187-196.
    Park EK,Shin YW,Lee HU,Kim SS,Lee YC,Lee BY,Kim DH(2005)Inhibitory effect of ginsenoside Rb1 and compound K on NO and prostaglandin E2 biosyntheses of RAW264.7 cells induced by lipopolysaccharide.Biol Pharm Bull 28:652-656.
    Paxinos G,Watson C(1997)The rat brain in stereotaxic coordinates.New York:Academic Press,USA.
    Radad K,Gille G,Moldzio R,Saito H,Ishige K,Rausch WD(2004)Ginsenosides Rb1 and Rg1 effects on survival and neurite growth of MPP+-affected mesencephalic dopaminergic cells.J Neural Transm 111:37-45.
    Rai SN,Birla H,Zahra W,Singh SS,Singh SP(2017)Immunomodulation of Parkinson’s disease using Mucuna pruriens(Mp).J Chem Neuroanat85:27-35.
    Rhule A,Navarro S,Smith JR,Shepherd DM(2006)Panax notoginseng attenuates LPS-induced pro-inflammatory mediators in RAW264.7 cells.J Ethnopharmacol 106:121-128.
    Rizek P,Kumar N,Jog MS(2016)An update on the diagnosis and treatment of Parkinson disease.CMAJ 188:1157-1165.
    Roussakis AA,Piccini P(2018)Molecular imaging of neuroinflammation in idiopathic parkinson’s disease.Int Rev Neurobiol 141:347-363.
    Sharma N,Sharma S,Nehru B(2017)Curcumin protects dopaminergic neurons against inflammation-mediated damage and improves motor dysfunction induced by single intranigral lipopolysaccharide injection.Inflammopharmacology 25:351-368.
    Smolinski AT,Pestka JJ(2003)Modulation of lipopolysaccharide-induced proinflammatory cytokine production in vitro and in vivo by the herbal constituents apigenin(chamomile),ginsenoside Rb(1)(ginseng)and parthenolide(feverfew).Food Chemical Toxicol 41:1381-1390.
    Song L,Xu MB,Zhou XL,Zhang DP,Zhang SL,Zheng GQ(2017)A preclinical systematic review of ginsenoside-rg1 in experimental parkinson’s disease.Oxid Med Cell Longev 2017:2163053.
    Sun XC,Ren XF,Chen L,Gao XQ,Xie JX,Chen WF(2016)Glucocorticoid receptor is involved in the neuroprotective effect of ginsenoside Rg1against inflammation-induced dopaminergic neuronal degeneration in substantia nigra.J Steroid biochem Mol Biol 155:94-103.
    Tansey MG,Goldberg MS(2010)Neuroinflammation in Parkinson’s disease:its role in neuronal death and implications for therapeutic intervention.Neurobiol Dis 37:510-518.
    Tansey MG,Romero-Ramos M(2019)Immune system responses in Parkinson’s disease:Early and dynamic.Eur J Neurosci 49:364-383.
    Tonges L,Metzdorf J,Zella S(2018)Parkinson’s disease and neuroinflammation-cellular pathology,mechanisms and therapeutic options.Fortschr Neurol Psychiatr 86:S10-S20.
    Wang GQ,Li DD,Huang C,Lu DS,Zhang C,Zhou SY,Liu J,Zhang F(2017)Icariin reduces dopaminergic neuronal loss and microglia-mediated inflammation in vivo and in vitro.Front Mol Neurosci 10:441.
    Wang Q,Liu Y,Zhou J(2015)Neuroinflammation in Parkinson’s disease and its potential as therapeutic target.Transl Neurodegener 4:19.
    Wang Y,Liu J,Zhang Z,Bi P,Qi Z,Zhang C(2011)Anti-neuroinflammation effect of ginsenoside Rbl in a rat model of Alzheimer disease.Neurosci Lett 487:70-72.
    Wang Y,Li Y,Yang W,Gao S,Lin J,Wang T,Zhou K,Hu H(2018)Ginsenoside Rb1 inhibit apoptosis in rat model of Alzheimer’s disease induced by Abeta1-40.Am J Transl Res 10:796-805.
    Woller SA,Ravula SB,Tucci FC,Beaton G,Corr M,Isseroff RR,Soulika AM,Chigbrow M,Eddinger KA,Yaksh TL(2016)Systemic TAK-242prevents intrathecal LPS evoked hyperalgesia in male,but not female mice and prevents delayed allodynia following intraplantar formalin in both male and female mice:the role of TLR4 in the evolution of a persistent pain state.Brain Behav immun 56:271-280.
    Xu M,Chen Q,Fan R,Wang J,Li Y(2019)Anti-inflammation effect of small molecule oligopeptides prepared from Panax ginseng C.A.Meyer in rats.Molecules 24:E858.
    Yu S,Zhou X,Li F,Xu C,Zheng F,Li J,Zhao H,Dai Y,Liu S,Feng Y(2017)Microbial transformation of ginsenoside Rb1,Re and Rg1 and its contribution to the improved anti-inflammatory activity of ginseng.Sci Rep7:138.
    Zeng XS,Geng WS,Jia JJ(2018)Neurotoxin-induced animal models of Parkinson disease:pathogenic mechanism and assessment.ASN Neuro doi:10.1177/1759091418777438.
    Zhang Y,Feng S,Nie K,Li Y,Gao Y,Gan R,Wang L,Li B,Sun X,Wang L,Zhang Y(2018a)TREM2 modulates microglia phenotypes in the neuroinflammation of Parkinson’s disease.Biochem Biophys Res Commun499:797-802.
    Zhang YL,Liu Y,Kang XP,Dou CY,Zhuo RG,Huang SQ,Peng L,Wen L(2018b)Ginsenoside Rb1 confers neuroprotection via promotion of glutamate transporters in a mouse model of Parkinson’s disease.Neuropharmacology 131:223-237.
    Zhao D,Zhang M,Yuan H,Meng C,Zhang B,Wu H(2018)Ginsenoside Rb1 protects against spinal cord ischemia-reperfusion injury in rats by downregulating the Bax/Bcl-2 ratio and caspase-3 and p-Ask-1 levels.Exp Mol Pathol 105:229-235.
    Zhou P,Xie W,Sun Y,Dai Z,Li G,Sun G,Sun X(2019)Ginsenoside Rb1and mitochondria:a short review of the literature.Mol Cell Probes 43:1-5.
    Zhu J,Jiang Y,Wu L,Lu T,Xu G,Liu X(2012)Suppression of local inflammation contributes to the neuroprotective effect of ginsenoside Rb1 in rats with cerebral ischemia.Neuroscience 202:342-351.
    Zhu JD,Wang JJ,Zhang XH,Yu Y,Kang ZS(2018)Panax ginseng extract attenuates neuronal injury and cognitive deficits in rats with vascular dementia induced by chronic cerebral hypoperfusion.Neural Regen Res13:664-672.