毛钩藤碱对6-羟基多巴胺诱导的帕金森病模型大鼠神经保护作用及机制研究

目的 探讨毛钩藤碱对帕金森病(Parkinson′s disease, PD)模型大鼠的神经保护作用及机制。方法 采用右侧中脑黑质致密部注射6-羟基多巴胺(6-hydroxydopamine, 6-OHDA)制备大鼠PD模型,将建模成功的PD大鼠随机分为模型组、美多芭组、毛钩藤碱低剂量和高剂量组,同时设置正常对照组,每组12只。给予相应药物灌胃干预,每天1次,连续1个月。采用旋转试验、转棒试验和Morris水迷宫试验检测大鼠行为学变化;HE染色法观察大鼠右侧中脑黑质病理学变化;高效液相色谱法(HPLC)检测大鼠右侧纹状体中多巴胺(dopamine, DA)水平;免疫组化法检测大鼠右侧中脑黑质中多巴胺能神经元标志物酪氨酸羟化酶(tyrosine hydroxylase, TH)表达;ELISA法检测大鼠右侧中脑黑质IL-6、IL-1β和TNF-α水平;Western blot检测大鼠右侧中脑黑质中NF-κB p65、NF-κB抑制因子α(IκBα)、p-IκBα等蛋白表达水平。结果 毛钩藤碱能显著改善PD大鼠行为学障碍及中脑黑质损伤,升高纹状体DA含量及中脑黑质中TH阳性神经元数量和Iκ...     

麦角甾醇对LPS诱导的肺损伤小鼠的抗炎作用研究

目的研究麦角甾醇对LPS诱导的小鼠急性肺损伤的抗炎作用研究。方法取50只雄性BABL/c小鼠随机分为5组:空白对照组、模型组、地塞米松组(2 mg/kg)、麦角甾醇低剂量组(20 mg/kg)、麦角甾醇高剂量组(40 mg/kg)。空白对照组和模型组按体积给予生理盐水,地塞米松组(2 mg/kg)、麦角甾醇组(20、40mg/kg)灌胃给予相应药物。给药1 h后,除空白对照组,其余各组小鼠气管滴注20μg LPS。检测小鼠肺干湿重比(W/D),肺泡灌洗液(BALF)中超氧化物歧化酶(SOD)水平、丙二醛(MDA)含量,血清与肺泡灌洗液炎性细胞因子(TNF-α、IL-1β、IL-6)水平。取各组小鼠肺组织做HE染色,并检测肺组织Rho、ROCK1、ROCK2、p-NF-κBP65、NF-κBP65、p-IκBα、IκBα蛋白表达。结果麦角甾醇20、40 mg/kg能显著提升LPS诱导的急性肺损伤小鼠血清SOD水平,降低MDA含量;改善血清及肺泡灌洗液炎症因子水平和肺组织病理学改变;降低肺组织Rho、ROCK1、ROCK2、p-NF-κBP65、p-IκBα蛋白表达。结论麦角甾醇对LPS诱导的急性肺损伤小鼠有保护作用,其作用可能与Rho/ROCK/NF-κB信号通路有关。     

神经酸对帕金森病模型小鼠运动障碍的缓解作用及机制研究

目的:研究神经酸对帕金森病(PD)模型小鼠运动障碍的缓解作用及机制。方法:将小鼠随机分为空白对照组(生理盐水)、模型组(生理盐水)、多巴丝肼片组(阳性对照,按左旋多巴计120 mg/kg)和神经酸低、中、高剂量组(20.0、40.0、80.0 mg/kg),每组10只。除空白对照组外,其余各组小鼠均建立PD模型。成模后,每天ig相应药物1次,连续14 d。末次给药后,观察各组小鼠行为学变化,采用高效液相色谱法测定小鼠脑纹状体内多巴胺(DA)及其代谢物二羟苯乙酸(DOPAC)、高香草酸(HVA)的含量。结果:与空白对照组比较,模型组小鼠爬杆时间延长、滚筒时间缩短、自发运动次数减少(P<0.05),脑纹状体内DA、DOPAC、HVA含量均降低(P<0.05)。与模型组比较,多巴丝肼片组和神经酸各剂量组小鼠爬杆时间缩短、滚筒时间延长(P<0.05),脑纹状体内DA、DOPAC、HVA含量均升高(P<0.05);多巴丝肼片组和神经酸高剂量组小鼠自发运动次数增加(P<0.05)。结论:神经酸可有效改善PD模型小鼠的运动障碍症状,其机制可能与增加脑纹状体内DA含量有关。     

类叶升麻苷对MPTP所致帕金森病小鼠模型的神经保护作用

目的研究类叶升麻苷在MPTP诱导的C57小鼠的帕金森病(PD)模型中的神经保护作用及机制。方法通过自主活动实验和滚筒实验研究动物的行为表现,通过高效液相电化学检测方法观察脑纹状体多巴胺的变化,通过脑黑质酪氨酸羟化酶(tyroxinehydroxylase,TH)免疫组化染色观察多巴胺能神经元的损伤程度。并对黑质纹状体进行α-突触核蛋白(α-synuclein)的免疫印迹分析以探讨药物作用机制。结果①经MPTP诱导的C57小鼠,其自主活动次数、滚筒运动潜伏期均低于对照组(P<0·01);纹状体多巴胺含量明显降低(P<0·01);多巴胺能神经元数量明显减少;黑质纹状体α-synuclein蛋白水平下降。②经类叶升麻苷(10、30mg·kg-1)预处理后能明显改善MPTP诱导的C57小鼠的行为学表现,增加脑内多巴胺递质的含量,增加多巴胺能神经元的数量,增加黑质纹状体α-synuclein蛋白水平。结论类叶升麻苷具有神经保护作用,能对抗MPTP诱导的C57小鼠PD模型中的神经损伤。其机制可能与上调α-synuclein蛋白水平有关。     

红景天苷调控PI3K/Akt信号通路对LPS诱导的BV2小胶质细胞的抗炎作用

目的探讨红景天苷对脂多糖(LPS)诱导的BV2小胶质细胞的抗炎作用及其机制。方法建立LPS诱导BV2小胶质细胞损伤模型。经不同浓度的红景天苷作用后,q PCR法检测细胞因子IL-6、IL-1β、TNF-αmRNA的表达; Western blot法检测Akt、p-Akt、核蛋白NF-κB p50的蛋白表达。PI3K抑制剂LY294002作用30 min,再经红景天苷作用后,检测Akt、p-Akt、核蛋白NF-κB p50、IL-6、IL-1β、TNF-α等指标。结果与模型组比较,红景天苷能够抑制BV2小胶质细胞IL-6、IL-1β、TNF-αmRNA的表达,促进p-Akt蛋白表达,抑制核蛋白NF-κB p50;经LY294002作用后,红景天苷对pAkt、核蛋白NF-κB p50、IL-6、IL-1β等作用不明显。结论红景天苷能够抑制LPS诱导的BV2小胶质细胞炎症反应,主要是通过激活PI3K/Akt信号通路,促进Akt的磷酸化,抑制NF-κB p50核转录,进而抑制细胞因子。     

雷公藤氯内酯醇对帕金森病大鼠多巴胺神经元的保护作用

目的探讨雷公藤氯内酯醇(T₄)对帕金森病(PD)大鼠多巴胺(DA)神经元的保护作用。方法采用线刀损毁大鼠内侧前脑束(MFB)制备部分损伤性PD模型,应用旋转行为测试行为学改变,HPLC-ECD检测纹状体DA含量,酪氨酸羟化酶(TH)免疫组化检测DA神经元的存活率,双抗夹心ELISA法检测损毁侧脑组织中细胞因子含量。结果T₄在较低剂量下(1 μg·kg^-1)能够改善安非他明(AMPH)诱发的PD大鼠异常旋转行为,其损伤侧纹状体DA含量和黑质致密部DA神经元存活率均比对照组增加,并能抑制脑内TNF-α和IL-2的异常升高。结论免疫抑制剂T₄对PD大鼠具有肯定的神经保护作用,其作用的发挥与T₄抵抗细胞因子在脑内过度升高而产生的毒性作用有关。     

中药提取物CTE对MPTP所致帕金森病小鼠模型的神经保护作用

目的:研究中药提取物CTE对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)所致帕金森病小鼠模型是否具有神经保护作用。方法:首先建立MPTP所致帕金森病小鼠模型,连续4 d给予MPTP(30 mg.kg-1,qd,ip)。用高低剂量(50和100 mg.kg-1,qd,ig)中药提取物CTE预处理MPTP所致帕金森模型小鼠。之后进行行为学检测,包括一般行为学检测和滚筒实验。最后,用HPLC-EC法测定纹状体中多巴胺(DA)及其代谢产物二羟苯乙酸(DOPAC)和高香草酸(HVA)含量。结果:在MPTP所致的帕金森病小鼠模型,中药提取物CTE能显著改善小鼠的行为能力;且能显著提高小鼠纹状体内多巴胺含量。结论:中药提取物CTE对MPTP所致帕金森病小鼠模型有神经保护作用。     

二苯乙烯苷对帕金森病模型大鼠行为学及多巴胺能神经元的影响

［摘要］　目的：观察何首乌主要有效成分二苯乙烯苷（TSG）对帕金森病（PD）模型大鼠行为学及脑内黑质多巴胺能神经元的影响。方法：采用6-羟基多巴胺（6-OHDA）单侧脑内黑质致密部（SNC）和中脑腹侧被盖区（VTA）两点注射法制备PD大鼠模型后，灌胃给予TSG低、中、高剂量 (50，100，200 mg&#8226;kg-1)，qd，连续5周。实验同时设正常对照组和美多芭阳性对照组（125 mg&#8226;kg-1），每组10只。用旋转行为检测及Rotarod试验观察各组大鼠的行为学变化，用免疫组化法观察大鼠脑黑质酪氨酸羟化酶(TH)阳性神经元的表达，用高效液相色谱-电化学法（HPLC-ECD）测定大鼠脑黑质多巴胺（DA）及3,4-二羟基苯乙酸（DOPAC）含量。结果：行为学检测结果表明，模型组大鼠旋转次数及运动协调性均低于正常对照组(P<0.01)，而TSG与美多芭给药可改善大鼠旋转行为及运动协调性。TH免疫组化检测结果显示，TSG处理组脑黑质部位损毁较轻，有较多残存的TH阳性细胞；而模型组黑质大部损毁，几乎无残存的TH阳性细胞；从HPLC-ECD测定结果可以看出，与正常对照组比较，模型组DA及DOPAC含量下降显著(P<0.05)，与模型组相比，TSG处理组脑内DA及DOPAC含量明显增加(P<0.05)。结论：TSG可改善PD大鼠的行为学改变，增加黑质-纹状体多巴胺及其代谢物含量并提高黑质多巴胺能神经元的残存率，提示TSG对PD可能具有一定的神经保护作用。     

MYR通过NF-κκB信号通路改善LPS诱导小鼠纹状体炎症反应

目的研究MYR对LPS诱导小鼠纹状体内神经炎症的作用及机制。方法雄性BALB/c小鼠随机分为正常对照组、LPS模型组、MYR 20和50 mg·kg~(-1)给药组。连续给予MYR 7 d,于末次给药后,模型组和给药组小鼠采用腹腔注射LPS5 mg·kg~(-1)诱导小鼠急性神经炎症的发生,LPS注射6 h后,ELISA法检测小鼠纹状体中IL-1β,IL-6,TNF-α,MCP-1以及ICAM-1等炎症因子的变化;Western印迹法检测小鼠纹状体中NF-κB信号通路相关蛋白的表达变化。结果与正常对照组比较,LPS诱导组小鼠纹状体中炎症因子IL-1β,IL-6,TNF-α,MCP-1以及ICAM-1显著增加(所有P<0.01),NF-κB信号通路相关蛋白表达显著增加。而给予MYR 20和50 mg·kg~(-1)可显著抑制LPS诱导的小鼠纹状体炎症因子的释放,抑制NF-κB,IκB蛋白的磷酸化,抑制胞浆内NF-κB的核转位。结论 MYR可有效抑制LPS诱导的神经炎症,保护小鼠纹状体中多巴胺神经元,其抗炎保护作用与抑制NF-κB信号通路密切相关。     

螺旋藻多糖对MPTP致多巴胺能神经元损伤的保护

目的探讨螺旋藻多糖(PSP)对MPTP制备的C57BL小鼠帕金森病(PD)模型多巴胺能神经元的保护作用。方法C57BL小鼠分5组:对照组、MPTP组、PSP高、中、低剂量预处理+MPTP组。用免疫组化、RT-PCR和HPLC-ECD方法检测黑质中酪氨酸羟化酶(TH)和多巴胺转运体(DAT)免疫反应阳性表达水平,TH和DAT mRNA表达水平及纹状体中多巴胺(DA)及其代谢产物3,4-二羟苯乙酸(DOPAC)和高香草酸(HVA)的含量。结果模型小鼠黑质中TH和DAT免疫阳性细胞数目减少,TH和DAT mRNA表达水平降低,纹状体中DA、DOPAC和HVA含量降低(P<0.01)。PSP预处理组与模型组相比,TH和DAT免疫阳性细胞数目增加,TH和DAT mRNA表达升高,DA、DOPAC和HVA含量升高(P<0.05)。结论PSP对MPTP所致C57BL小鼠PD模型的多巴胺能神经元有保护作用。     

红景天苷对脂多糖诱导的支气管上皮细胞损伤的保护作用

目的：探讨红景天苷对脂多糖（LPS）诱导的正常人支气管上皮细胞（NHBE）损伤的保护作用及机制。方法：培养NHBE细胞,调整其细胞的密度为每毫升含有1＆#215;104个,给予不同浓度的红景天苷（3.3、10、30μg·mL-1）作用24 h,MTT检测细胞的活力,观察红景天苷在正常的情况下对人支气管上皮细胞的影响。在此基础上,以终浓度为2μg·mL-1的LPS作用NHBE细胞6 h,诱导其损伤,然后给予红景天苷24 h,MTT检测细胞活力,收集上清液,ELISA试剂盒检测炎症因子IL-6（白介素-6）、IL-1β（白介素-1β）、TNF-α（肿瘤坏死因子-α）的水平,收集细胞,用Western blot方法测定细胞中的蛋白NF-κB（核因子）、MAPK（丝裂原活化蛋白激酶）的表达。结果：在正常条件下,红景天苷对人支气管上皮细胞的活力无影响,没有损伤作用。红景天苷可明显降低培养液中IL-6、IL-1β、TNF-α的水平（P〈0.01）,降低细胞中NF-κB、MAPK的表达（P〈0.01）。结论：红景天苷可以通过降低IL-6、 IL-1β、TNF-α的水平来减轻LPS诱导的NHBE细胞的损伤,减轻炎症反应。     

Crocetin ester improves myocardial ischemia via Rho/ROCK/NF-κB pathway

Crocetin ester (CE) is the active ingredient of Crocus sativus L. stigmas and Gardenia jasminoides Ellis fruit. The main purpose of the present study was to investigate the protective effect of CE on isoproterenol (ISO)-induced acute myocardial ischemia (AMI) through Rho/ROCK/NF-κB pathway and explore its underlying mechanism. Administration of CE (25 and 50 mg/kg) could significantly reduce the serum contents of pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6). In addition, pretreatment with CE attenuated the contents of creatine kinase (CK), malondialdehyde (MDA) and the activities of lactate dehydrogenase (LDH), superoxide dismutase (SOD) in serum. Treatment with CE also improved the histopathological alteration and decreased the ST elevation. Furthermore, CE could ameliorate the cardiac expressions of Cu, Zn-superoxide dismutase (SOD1), MDA5, Rho, ROCK, p-IκB and p-NF-κBp65 in ISO-induced rats. It was assumed that CE might be a new therapeutic candidate for the treatment of AMI possibly through the inhibition of Rho/ROCK/NF-κB pathway.     

MPTP诱导野生型和A53T转基因型小鼠帕金森病模型纹状体中神经递质变化

目的 探究帕金森病(Parkinson's disease,PD)小鼠模型纹状体中多巴胺等神经递质的变化.方法 通过腹腔注射丙磺舒(probenecid)、皮下注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)制备PD野...     

Coadministration of β‐asarone and levodopa increases dopamine in rat brain by accelerating transformation of levodopa: A different mechanism from Madopar

The aim of the present study was to investigate the effect of coadministration of β‐asarone and levodopa (l ‐dopa) on increasing dopamine (DA) in the striatum of healthy rats. Rats were randomly divided into four groups: (i) a normal group, administered normal saline; (ii) a Madopar group, administered 75 mg/kg Madopar (l ‐dopa : benserazide, 4 : 1); (iii) an l ‐dopa group, administered 60 mg/kg l ‐dopa; and (iv) a group coadministered 15 mg/kg β‐asarone and 60 mg/kg l ‐dopa. All drugs (or normal saline) were administered intragastrically twice a day for 7 days. Then, plasma and striatum concentrations of DA, l ‐dopa, 5‐hydroxytryptamine (5‐HT), homovanillic acid (HVA), 3,4‐dihydroxyphenylacetic acid (DOPAC), tyrosine hydroxylase (TH), catechol‐O‐methyltransferase (COMT) and monoamine oxidase B (MAO‐B) were determined. In the group coadministered β‐asarone and l ‐dopa, there was a decline in plasma and striatal concentrations of l ‐dopa; however, DA and DOPAC concentrations increased in the striatum and plasma and plasma HVA concentrations increased, whereas there was no significant change in striatal levels. Concentrations of 5‐HT in the striatum and plasma were similar in the coadministered and Madopar‐treated groups. In addition, plasma and striatal COMT levels decreased after coadministration of β‐asarone and l ‐dopa, whereas there were no significant differences in MAO‐B concentrations among groups. Furthermore, coadministration of β‐asarone and l ‐dopa increased plasma TH concentrations. Altogether, β‐asarone affects the conversion of l ‐dopa to DA by modulating COMT activity and DA metabolism. The mechanism of coadministration is different from that of Madopar in Parkinson's disease (PD) treatment. Thus, the coadministration of β‐asarone and l ‐dopa may be beneficial in the treatment of PD.     

Salidroside attenuates apoptosis in ischemic cardiomyocytes: a mechanism through a mitochondria-dependent pathway

In the present study, we investigated cardioprotective effects of salidroside, isolated from Rhodiola rosea L, on oxygen-glucose deprivation (OGD)-induced cardiomyocyte death and ischemic injury evoked by acute myocardial infarction (AMI) in rats. Pretreatment with salidroside notably ameliorated cell viability losses in a dose-dependant manner and in parallel it alleviated morphologic injury detected by electron microscopy. Mechanistically, diminished OGD-induced cardiomyocyte apoptosis was shown in salidroside-pretreated cardiomyocytes, in accordance with minimal reactive oxygen species (ROS) burst. Moreover, salidroside markedly upregulated the Bcl-2/Bax ratio and preserved mitochondrial transmembrane potential (ΔΨm). Salidroside administration also inhibited myocardial apoptosis in AMI rats by increasing phosphorylation of Akt and decreasing activation of caspase-3. These findings suggest that salidroside reduced ischemia-mediated myocardial damage. Salidroside therefore has potential to be a promising drug for preventing and treating myocardial ischemic diseases.     

Salidroside suppresses group 2 innate lymphoid cell-mediated allergic airway inflammation by targeting IL-33/ST2 axis

Salidroside, an active component extracted from Rhodiola rosea, has been reported to inhibit allergic asthma. However, its mechanism has not been fully elucidated. Group 2 innate lymphoid cells (ILC2s) accumulate in the lung and cooperate with other cells to drive type 2 inflammation stimulated by inhaled allergens. The study aims to explore the suppressive effect of salidroside on ILC2s and IL-33/IL-33R (ST2) axis in allergic airway inflammation. The ovalbumin (OVA)-sensitized/challenged mice were established. Airway eosinophil recruitment, increased total IgE in the serum and type 2 cytokines IL-4, IL-5, and IL-13 in the bronchoalveolar lavage fluids and lung tissues were identified in the OVA-induced mice model, all of which were inhibited by pretreatment with different doses of salidroside. Moreover, salidroside suppressed lung total ILC2 and ST2-expressing ILC2 accumulation, lung IL-33 and ST2 expressions in mice. In vitro, OVA could induce IL-33 expression in BEAS-2B cells, which was also effectively inhibited by salidroside. This study firstly reveals salidroside as a potential therapeutic drug for allergic asthma by inhibiting ILC2-mediated airway inflammation via targeting IL-33/ST2 axis.     

β‐asarone and levodopa co‐administration increase striatal dopamine level in 6‐hydroxydopamine induced rats by modulating P‐glycoprotein and tight junction proteins at the blood‐brain barrier and promoting levodopa into the brain

Levodopa (L‐dopa) is widely considered as one of the most effective drug constituents in the treatment of Parkinson's disease (PD), but the blood‐brain barrier (BBB) permeability of L‐dopa is <5%, which causes low efficacy. Neuroprotective effects of β‐asarone on 6‐hydroxydopamine (6‐OHDA)‐induced PD rats were demonstrated by our previous studies. Co‐administration of β‐asarone and L‐dopa has not been explored until being investigated on PD rats in this study. PD rats were divided into four groups: untreated, L‐dopa‐treated, β‐asarone‐treated and co‐administered‐treated groups. All of the treatments were administered to the rats twice per day for 30 days. The L‐dopa, dopamine (DA), 3,4‐dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), S100β and neuron‐specific enolase (NSE) levels were subsequently determined. The P‐glycoprotein (P‐gp), zonula occludens‐1 (ZO‐1), claudin‐5, occludin and actin expression was also assessed in cortex. Changes in BBB ultrastructure were observed using transmission electron microscopy. Our results showed that the co‐administered treatment increased levels of L‐dopa, DA, DOPAC and HVA in striatum, and S100β in plasma, but down‐regulated NSE, P‐gp, ZO‐1, occludin, actin and claudin‐5 in cortex. Crevices were observed between capillary endothelial cells at intercellular tight junction of the striatum in co‐administered‐treated group, while the endothelial cells in untreated group were tightly jointing each other. In addition, the correlations of L‐dopa or DA and P‐gp or tight junction proteins respectively were significantly negative in co‐administered‐ and β‐asarone‐treated groups. These findings suggest that co‐administered treatment may enhance the L‐dopa BBB permeability and attenuate brain injury, which may be beneficial to PD treatment.     

复方海蛇胶囊对帕金森病小鼠的作用

目的：研究复方海蛇胶囊对1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的帕金森病小鼠的作用。方法：采用C57BL/6J小鼠腹腔注射MPTP制备帕金森病动物模型,给药14 d后观察各组小鼠行为活动变化,利用高效液相-电化学检测器检测纹状体内多巴胺（DA）及其代谢产物二羟基苯乙酸（DOPAC）和高香草酸（HVA）等的含量变化。结果：复方海蛇胶囊可以显著对抗MPTP对黑质及纹状体的损伤作用,明显改善帕金森病的症状;与模型组相比,复方海蛇胶囊治疗后可以显著提高小鼠纹状体内DA、DOPAC和HVA的含量（P〈0.01）。结论：复方海蛇胶囊通过保护多巴胺能神经元,提高纹状体内DA的含量,改善帕金森病的症状。     

丙戊酸联合碳酸锂治疗帕金森病模型小鼠的实验研究

目的探讨丙戊酸联合碳酸锂(MPTP)对帕金森病(PD)小鼠模型的治疗作用。方法60只小鼠腹腔注射MPTP制作PD小鼠模型后均分为五组,分别予以丙戊酸联合碳酸锂(A组)、碳酸锂(B组)、丙戊酸(C组)和生理盐水(D组)处理。采用免疫组织化学及Western blot方法检测PD相关标志物酪氨酸羟化酶(TH),高效液相色谱-电化学检测仪(HPLC-ECD)检测纹状体单胺类递质变化。结果与空白对照组(E组,12只)比较,D组TH阳性细胞数明显减少,多巴胺(DA)及其代谢产物二羟苯乙酸(DOPAC)明显下降。A、B、C组纹状体TH表达较D、E组明显增多,尤其A组明显;A组DOPAC含量较D组明显升高(P<0.05)。结论丙戊酸联合碳酸锂对MPTP诱导的PD小鼠有明显的治疗作用。