Sirt1通过促进海马中小胶质细胞向M2型转化改善慢性温和不可预知性应激模型小鼠的抑郁样行为
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摘要
目的探讨沉默信息调节因子2相关酶1 (silent mating-type information regulation 2 homolog 1,Sirt1)对小鼠抑郁样行为的影响以及相关的分子机制。方法共纳入60只8周龄C57BL/6雄性小鼠,利用慢性温和不可预知性应激(chronic mild unpredictable stress,CUMS)建立抑郁症小鼠模型。实验分为Control组(n=20)、CUMS组(n=20)和CUMS+Sirt1组(n=20,小鼠双侧海马注射Sirt1激动剂SRT2104各5μmol)。采用行为学、免疫荧光、流式细胞术以及Western blot等方法,对小鼠的抑郁样行为、小胶质细胞表型、GSK3β/PTEN信号通路进行检测。结果与CUMS组比较,CUMS+Sirt1组小鼠对糖水的偏好程度明显增加(P<0.01),同时在强迫游泳实验与悬尾实验中的不动时间明显降低(P<0.01);与CUMS组比较,虽然CUMS+Sirt1组小鼠脑内总的小胶质细胞数量无显著变化,但脑内的M1型小胶质细胞数量明显减少(P<0.01),M2型小胶质细胞数量明显增加(P<0.01),M1/M2的比率也明显减少(P<0.01);相对于CUMS组,CUMS+Sirt1组小鼠海马中Sirt1、P-GSK3β和P-PTEN蛋白表达也显著增高(P<0.01)。结论 Sirt1通过促进抑郁症小鼠模型海马中的小胶质细胞向M2型转化进而改善小鼠的抑郁样行为。
Objective To explore the effect of silent mating-type information regulation 2 homolog 1(Sirt1) on depression-like behaviors in mice and the underlying molecular mechanism. Methods Forty 8-week-old male C57 BL/6 mice were subjected to chronic mild unpredictable stress(CUMS) to establish models of depression. The mouse models were randomized equally into CUMS group(n=20) and CUMS+Sirt1 group(n=20), and the mice in the latter group were injected with 5 μmol SRT2104, a Sirt1 agonist, in the bilateral hippocampus. With another 20 mice without CUMS as the control group, the mice in the 2 CUMS groups were evaluated for depression-like behaviors; the hippocampal microglia number and phenotypes in the mice were assessed by immunofluorescence staining and flow cytometry, and Western blotting was used to detect the expression of Sirt1, P-GSK3β and P-PTEN in the hippocampus. Results Compared with those in CUMS group, the mice in CUMS+Sirt1 group showed a significantly increased sucrose preference(P<0.01) with obviously decreased immobility time in forced swimming test(P<0.01) and tail suspension test(P<0.01). Although the number of hippocampal microglia did not differ significantly between the 2 CUMS groups(P>0.05), the number of M1 phenotype microglia was decreased and that of M2 phenotype microglia was increased significantly in CUMS+Sirt1 group(P<0.01); the expression of Sirt1, P-GSK3β and P-PTEN in the hippocampus were increased significantly in CUMS+Sirt1 group compared 2 those in CUMS group(P<0.01). Conclusion Sirt1 ameliorates CUMS-induced depression-like behaviors in mice by promoting the transformation of microglia into M2 phenotype.
Objective To explore the effect of silent mating-type information regulation 2 homolog 1(Sirt1) on depression-like behaviors in mice and the underlying molecular mechanism. Methods Forty 8-week-old male C57 BL/6 mice were subjected to chronic mild unpredictable stress(CUMS) to establish models of depression. The mouse models were randomized equally into CUMS group(n=20) and CUMS+Sirt1 group(n=20), and the mice in the latter group were injected with 5 μmol SRT2104, a Sirt1 agonist, in the bilateral hippocampus. With another 20 mice without CUMS as the control group, the mice in the 2 CUMS groups were evaluated for depression-like behaviors; the hippocampal microglia number and phenotypes in the mice were assessed by immunofluorescence staining and flow cytometry, and Western blotting was used to detect the expression of Sirt1, P-GSK3β and P-PTEN in the hippocampus. Results Compared with those in CUMS group, the mice in CUMS+Sirt1 group showed a significantly increased sucrose preference(P<0.01) with obviously decreased immobility time in forced swimming test(P<0.01) and tail suspension test(P<0.01). Although the number of hippocampal microglia did not differ significantly between the 2 CUMS groups(P>0.05), the number of M1 phenotype microglia was decreased and that of M2 phenotype microglia was increased significantly in CUMS+Sirt1 group(P<0.01); the expression of Sirt1, P-GSK3β and P-PTEN in the hippocampus were increased significantly in CUMS+Sirt1 group compared 2 those in CUMS group(P<0.01). Conclusion Sirt1 ameliorates CUMS-induced depression-like behaviors in mice by promoting the transformation of microglia into M2 phenotype.
引文
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[2] LEE Y,SUBRAMANIAPILLAI M,BRIETZKE E,et al.Anti-cytokine agents for anhedonia:targeting inflammation and the immune system to treat dimensional disturbances in depression[J].Ther Adv Psychopharmacol,2018,8(12):337-348.DOI:10.1177/2045125318791944.
[3] ZHANG L J,ZHANG J Q,YOU Z L.Switching of the microglial activation phenotype is a possible treatment for depression disorder[J].Front Cell Neurosci,2018,12:306.DOI:10.3389/fncel.2018.00306.
[4] CONVERGE CONSORTIUM.Sparse whole-genome sequenc-ing identifies two loci for major depressive disorder[J].Nature,2015,523(7562):588-591.DOI:10.1038/nature 14659.
[5] TANG C Z,YANG J T,LIU Q H,et al.Up-regulated miR-192-5p expression rescues cognitive impairment and restores neural function in mice with depression via the Fbln2-mediated TGF-β1 signaling pathway[J].FASEB J,2019,33(1):606-618.DOI:10.1096/fj.201800210RR.
[6] SU W J,ZHANG Y,CHEN Y,et al.NLRP3 gene knockout blocks NF-κB and MAPK signaling pathway in CUMS-induced depression mouse model[J].Behav Brain Res,2017,322(Pt A):1-8.DOI:10.1016/j.bbr.2017.01.018.
[7] LIU L,ZHANG Q,CAI Y L,et al.Resveratrol counteracts lipopolysaccharide-induced depressive-like behaviors via enhanced hippocampal neurogenesis[J].Oncotarget,2016,7(35):56045-56059.DOI:10.18632/oncotarget.11178.
[8] NI Y F,WANG H,GU Q Y,et al.Gemfibrozil has antidepressant effects in mice:Involvement of the hippocampal brain-derived neurotrophic factor system[J].J Psychopharmacol (Oxford),2018,32(4):469-481.DOI:10.1177/0269881118762072.
[9] WANG Y H,XU J J,LIU Y,et al.TLR4-NF-κB signal involved in depressive-like behaviors and cytokine expression of frontal cortex and hippocampus in stressed C57BL/6 and ob/ob mice[J].Neural Plast,2018,2018:7254016.DOI:10.1155/2018/7254016.
[10] TAN S J,WANG Y,CHEN K,et al.Ketamine alleviates depressive-like behaviors via down-regulating inflammatory cytokines induced by chronic restraint stress in mice[J].Biol Pharm Bull,2017,40(8):1260-1267.DOI:10.1248/bpb.b17-00131.
[11] DEL GRANDE DA SILVA G,WIENER C D,BARBOSA L P,et al.Pro-inflammatory cytokines and psychotherapy in depression:Results from a randomized clinical trial[J].J Psychiatr Res,2016,75:57-64.DOI:10.1016/j.jpsychires.2016.01.008.
[12] ZOU W,FENG R J,YANG Y.Changes in the serum levels of inflammatory cytokines in antidepressant drug-na?ve patients with major depression[J].PLoS ONE,2018,13(6):e0197267.DOI:10.1371/journal.pone.0197267.
[13] CHERRY J D,OLSCHOWKA J A,O’BANION M K.Neuroinflammation and M2 microglia:the good,the bad,and the inflamed[J].J Neuroinflammation,2014,11:98.DOI:10.1186/1742-2094-11-98.
[14] YANG J P,ZHAO Y Y,ZHANG L,et al.RIPK3/MLKL-mediated neuronal necroptosis modulates the M1/M2 polarization of microglia/macrophages in the ischemic cortex[J].Cerebral Cortex,2018,28(7):2622-2635.DOI:10.1093/cercor/bhy089.
[15] ZHOU K,ZHONG Q,WANG Y C,et al.Regulatory T cells ameliorate intracerebral hemorrhage-induced inflammatory injury by modulating microglia/macrophage polarization through the IL-10/GSK3β/PTEN axis[J].J Cereb Blood Flow Metab,2017,37(3):967-979.DOI:10.1177/0271678X16648712.
[16] WANG G H,SHI Y J,JIANG X Y,et al.HDAC inhibition prevents white matter injury by modulating microglia/macrophage polarization through the GSK3β/PTEN/Akt axis[J].Proc Natl Acad Sci U S A,2015,112(9):2853-2858.DOI:10.1073/pnas.1501441112.
[17] ZHANG J Q,XIE X F,TANG M M,et al.Salvianolic acid B promotes microglial M2-polarization and rescues neurogenesis in stress-exposed mice[J].Brain Behav Immun,2017,66:111-124.DOI:10.1016/j.bbi.2017.07.012.
[18] ZHAO Q Y,WU X H,YAN S,et al.The antidepressant-like effects of pioglitazone in a chronic mild stress mouse model are associated with PPARγ-mediated alteration of microglial activation phenotypes[J].J Neuroinflammation,2016,13(1):259.DOI:10.1186/s12974-016-0728-y.
[19] BUHRMANN C,BUSCH F,SHAYAN P,et al.Sirtuin-1 (SIRT1) is required for promoting chondrogenic differentiation of mesenchymal stem cells[J].J Biol Chem,2014,289(32):22048-22062.DOI:10.1074/jbc.M114.568790.
[20] CHEN X R,CHEN C N,FAN S N,et al.Omega-3 polyunsaturated fatty acid attenuates the inflammatory response by modulating microglia polarization through SIRT1-mediated deacetylation of the HMGB1/NF-κB pathway following experimental traumatic brain injury[J].J Neuroinflammation,2018,15(1):116.DOI:10.1186/ s12974-018-1151-3.
[21] WU H,WU J,ZHOU S,et al.SRT2104 attenuates diabetes-induced aortic endothelial dysfunction via inhibition of P53[J].J Endocrinol.2018,237(1):1-14.DOI:10.1530/JOE-17-0672.