PDTC对AD模型小鼠海马区Tau蛋白及胶质细胞的影响
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摘要
目的:探讨吡咯烷二硫代甲酸(pyrrolidine dithiocarbamate,PDTC)对AD模型小鼠海马区Tau蛋白及胶质细胞的影响。方法:40只雄性昆明小鼠随机分为对照组、Aβ组、Aβ+6 mg/L PDTC组和Aβ+8 mg/L PDTC组,每组各8只动物。Aβ组小鼠通过单侧海马注射Aβ1-42进行造模,对侧侧脑室注射生理盐水;PDTC处理组小鼠单侧海马注射Aβ1-42,对侧侧脑室内注射PDTC进行干预。采用免疫组织化学方法检测小鼠海马区S100β的表达情况,银染法检测小鼠海马齿状回Tau蛋白的表达情况,采用Western Blot法检测小鼠大脑海马区CD11b、p-Tau(S404)及GFAP蛋白的表达情况。结果:免疫组织化学实验结果显示:与对照组比较,Aβ组小鼠整个海马区星形胶质细胞标记物S100β阳性细胞的表达显著增加(P<0.01);给予不同浓度PDTC处理后,S100β阳性细胞的表达减少(P<0.05)。银染法实验结果显示:与对照组比较,Aβ组小鼠海马齿状回Tau蛋白阳性细胞的表达显著增加(P<0.01);给予不同浓度PDTC处理后,Tau蛋白阳性细胞的表达减少(P<0.05)。Western Blot实验结果显示:与对照组比较,Aβ组小鼠大脑海马区CD11b、p-Tau(S404)及GFAP蛋白表达均增多(P<0.05)。给予不同浓度PDTC处理后,CD11b蛋白表达显著增多(P<0.01);p-Tau(S404)和GFAP蛋白表达均减少(P<0.05)。结论:PDTC可减少AD模型小鼠海马区Tau蛋白与星形胶质细胞表达;增加小胶质细胞的表达,其相关机制仍需进一步的研究。
Objective:This study aims to investigate effects of pyrrolidine dithiocarbamate(PDTC) on Tau protein and glial cells in the hippocampus of AD model mice.Methods:Forty Male KM mice were randomly divided into four groups,including the control group,Aβ group,Aβ plus 6 mg/L PDTC group and Aβ plus 8 mg/L PDTC group,eight mice each group.The Aβ group mice were injected with Aβ1-42 through unilaterally hippocampal injection,and saline was injected into contralateral ventricles.For PDTC treatment group,Aβ1-42 was injected into unilateral hippocampus,and PDTC was injected into lateral ventricle.Immunohistochemical method was used to detect the expression of S100β in the hippocampus of mice.The expression of Tau protein in the dentate gyrus of hippocampus of mice was detected by silver staining method.The expressions of CD11 b,p-Tau(S404) and GFAP protein in the brain of mice were detected by Western Blot method.Results:The results of immunohistochemistry showed that the expression of astrocyte marker S100βpositive cells were significantly increased in the hippocampus of Aβ mice(P < 0.01) when compared to the control.Afterthe treatment of different concentrations of PDTC,the expressions of S100β positive cells decreased in the hippocampus of mice(P < 0.05).The results of silver staining showed that the expression of Tau protein positive cells were significantly increased in the dentate gyrus of hippocampus of Aβ mice(P < 0.01) when compared to the control.After the treatment of different concentrations of PDTC,the expressions of Tau protein positive cells decreased(P < 0.05).The Western Blot results showed that compared with the control group,the expressions of CD11 b,S404 and GFAP protein in the brain of Aβ group increased(P < 0.05).After treatment with different concentrations of PDTC,the expression of CD11 b protein was increased significantly(P < 0.01).The expressions of p-Tau(S404) and GFAP protein were decreased(P <0.05).Conclusion:PDTC can reduce the expressions of Tau protein and astrocytes,and increase the expression of microglia in the hippocampus of AD model mice,further research is still needed in the related mechanisms.
Objective:This study aims to investigate effects of pyrrolidine dithiocarbamate(PDTC) on Tau protein and glial cells in the hippocampus of AD model mice.Methods:Forty Male KM mice were randomly divided into four groups,including the control group,Aβ group,Aβ plus 6 mg/L PDTC group and Aβ plus 8 mg/L PDTC group,eight mice each group.The Aβ group mice were injected with Aβ1-42 through unilaterally hippocampal injection,and saline was injected into contralateral ventricles.For PDTC treatment group,Aβ1-42 was injected into unilateral hippocampus,and PDTC was injected into lateral ventricle.Immunohistochemical method was used to detect the expression of S100β in the hippocampus of mice.The expression of Tau protein in the dentate gyrus of hippocampus of mice was detected by silver staining method.The expressions of CD11 b,p-Tau(S404) and GFAP protein in the brain of mice were detected by Western Blot method.Results:The results of immunohistochemistry showed that the expression of astrocyte marker S100βpositive cells were significantly increased in the hippocampus of Aβ mice(P < 0.01) when compared to the control.Afterthe treatment of different concentrations of PDTC,the expressions of S100β positive cells decreased in the hippocampus of mice(P < 0.05).The results of silver staining showed that the expression of Tau protein positive cells were significantly increased in the dentate gyrus of hippocampus of Aβ mice(P < 0.01) when compared to the control.After the treatment of different concentrations of PDTC,the expressions of Tau protein positive cells decreased(P < 0.05).The Western Blot results showed that compared with the control group,the expressions of CD11 b,S404 and GFAP protein in the brain of Aβ group increased(P < 0.05).After treatment with different concentrations of PDTC,the expression of CD11 b protein was increased significantly(P < 0.01).The expressions of p-Tau(S404) and GFAP protein were decreased(P <0.05).Conclusion:PDTC can reduce the expressions of Tau protein and astrocytes,and increase the expression of microglia in the hippocampus of AD model mice,further research is still needed in the related mechanisms.
引文
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[3]Cai Z,Hussain MD,Yan LJ.Microglia,neuroinflammation,and Aβin Alzheimer’s disease[J].Int J Neurosci,2014,124:307-321.
[4]Asai H,Ikezu S,Tsunoda S,et al.Depletion of microglia and inhibition of exosome synthesis halt tau propagation[J].Nat Neurosci,2015,18:1584-1593.
[5]Phillips EC,Croft CL,Kurbatskaya K,et al.Astrocytes and neuroinflammation in Alzheimer’s Disease[J].Biochem Soc Trans,2014,42:1321-1325.
[6]Yasuda Y,Tateishi N,Shimoda T,et al.Relationship between S100βand GFAP expression in astrocytes during infarction and glial scar formation after mild transient ischemia[J].Brain Res,2004,1021:20-31.
[7]González-Reyes RE,Nava-Mesa MO,Vargas-Sánchez K,et al.Involvement of astrocytes in Alzheimer’s Disease from a neuroinflammatory and oxidative stress perspective[J].Front Mol Neurosci,2017,10:427.
[8]Yata K,Oikawa S,Sasaki R,et al.Astrocytic neuroprotection through induction of cytoprotective molecules;a proteomic analysis of mutant P301S tau-transgenic mouse[J].Brain Res,2011,1410:12-23.
[9]Sato-Harada R,Okabe S,Umeyama T,et al.Microtubule-associated proteins regulate microtubule function as the track for intracellular membrane organelle transports[J].Cell Struct Funct,1996,21:283-295.
[10]Roberson ED,Scearce-Levie K,Palop JJ,et al.Reducing endogenous tau ameliorates amyloid beta-induced deficits in an Alzheimer’s disease mouse model[J].Science,2007,316:750-754.
[11]Kolarova M,Garca-Sierra F,Bartos A,et al.Structure and pathology of Tau protein in Alzheimer disease[J].Int J Alzheimers Dis,2012,2012:731526.
[12]Liddell JR,Lehtonen S,Duncan C,et al.Pyrrolidine dithiocarbamate activates the Nrf2 pathway in astrocytes[J].J Neuroinflammation,2016,13:49.
[13]Abd-El-Fattah MA,Abdelakader NF,Zaki HF.Pyrrolidine dithiocarbamate protects against scopolamine-induced cognitive impairment in rats[J].Eur J Pharmacol,2014,723:330-338.
[14]Helmfors L,Boman A,Civitelli L,et al.Protective properties of lysozyme onβ-amyloid pathology:implications for Alzheimer disease[J].Neurobiol Dis,2015,83:122-133.
[15]Herrup K.The case for rejecting the amyloid cascade hypothesis[J].Nat Neurosci,2015,18:794-799.
[16]Stepanichev MY,Zdobnova IM,Yakovlev AA,et al.Effects of tumor necrosis factor-alpha central administration on hippocampal damage in rat induced by amyloid beta-peptide(25-35)[J].J Neurosci Res,2003,71:110-120.
[17]Yang J,Yu XX,Abulaiti A,et al.Correlation between nuclear factorκB activity and pulmonary artery pressure in a rat high pulmonary blood flow model[J].Exp Ther Med,2015,9:543-546.
[18]Gomez-Nicola1 D,Perry VH.Microglial dynamics and role in the healthy and diseased brain:A paradigm of functional plasticity[J].Neuroscientist,2015,21:169-184.
[19]Steinberg S.Variant of TREM2 associated with the risk of Alzheimer’s Disease[J].New Engl J Med,2012,368:107-116.
[20]Hayden MS,Ghosh S.Shared principles in NF-kappa B signaling[J].Cell,2008,132:344-362.
[21]Maeda S,Kamata H,Luo J L,et al.IKKbeta couples hepatocyte death to cytokine-driven compensatory proliferation that promotes chemical hepatocarcinogenesis[J].Cell,2005,121:977-990.
[22]Zhao YH,Bhattacharjee S,Jones BM,et al.Regulation of TREM2expression by an NF-κB-sensitive mi RNA-34a[J].Neuroreport,2013,24:318-323.
[23].Jay TR,Miller CM,Cheng PJ,et al.TREM2 deficiency eliminates TREM2+inflammatory macrophages and ameliorates pathology in Alzheimer’s disease mouse models[J].J Exp Med.2015,212:287-295.
[24]Avila-Mu1oz E,Arias C.When astrocytes become harmful:functional and inflammatory responses that contribute to Alzheimer’s disease[J].Ageing Res Rev,2014,18:29-40.