从NLRP3炎症小体途径探讨竹节参总皂苷改善自然衰老大鼠认知功能衰退的作用机制
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摘要
探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠认知功能衰退的改善作用及机制。18月龄SD雄性大鼠随机分为3组,即自然衰老组、竹节参总皂苷10 mg·kg~(-1)和30 mg·kg~(-1)剂量组,每组10只。竹节参总皂苷给药组大鼠从18月龄开始分别灌胃给予10,30 mg·kg~(-1)竹节参总皂苷,自然衰老组大鼠灌胃等量的生理盐水。灌胃持续6个月,每周连续灌胃6 d,停药1 d,另取10只6月龄的大鼠作为青年对照组。采用旷场、新物体认知和Morris水迷宫方法检测各组大鼠认知功能的改变,场电位记录法检测各组大鼠海马CA1区突触传递长时程增强(LTP)的变化。Western blot法检测各组大鼠NLRP3炎症小体相关蛋白NLRP3,ASC,caspase-1的表达变化以及突触可塑性相关蛋白Glu A1,Glu A2和学习记忆相关蛋白CAMKⅡ,CREB及其磷酸化表达的变化。竹节参总皂苷可改善衰老大鼠的认知功能衰退,减轻衰老大鼠海马CA1区LTP的受损程度,减少衰老大鼠NLRP3炎症小体相关蛋白NLRP3,ASC,caspase-1的表达水平。同时,竹节参总皂苷可增强突触可塑性相关蛋白AMPA受体亚型Glu A1和Glu A2的膜表达,增强衰老大鼠学习记忆相关蛋白CAMKⅡ,CREB的磷酸化表达水平。竹节参总皂苷可改善衰老大鼠认知功能衰退,其机制可能与其调节NLRP3炎症小体,进而调节AMPA受体膜表达,增强学习记忆相关蛋白CAMKⅡ和CREB的磷酸化表达有关。
The aim of this paper was to investigate the effect of total saponins from Panax japonicus( SPJ) on cognitive decline of natural aging rats and its mechanism. Thirty male SD rats of eighteen month old were randomly divided into three groups: aged group,10 mg·kg~(-1) SPJ-treated group and 30 mg·kg~(-1) SPJ-treated group. The SPJ-treated groups were given SPJ at the dosages of 10 mg·kg~(-1) and 30 mg·kg~(-1),respectively,from the age of 18 to 24 months. Aged group were lavaged the same amount of saline,10 six-month-old rats were used as control group,with 10 rats in each group. The open field test,novel object recognition and Morris water maze were performed to detect the changes of cognitive function in each group. The changes of synaptic transmission of long-term potentiation( LTP) in hippocampal CA1 region were detected by field potential recording. Western blot was used to detect the protein levels of NLRP3,ASC,caspase-1 and the changes of Glu A1,Glu A2,CAMKⅡ,CREB and phosphorylation of CAMKⅡ,CREB in each group.The results showed that SPJ could improve the decline of cognitive function in aging rats,reduce the damage of LTP in the hippocampal CA1 region of aged rats,and decrease the expression of NLRP3,ASC,caspase-1 in aging rats. At the same time,SPJ could enhance the membrane expression of AMPA receptor( Glu A1 and Glu A2),and increase the expression of p-CAMKⅡand p-CREB in aging rats.SPJ could improve cognitive decline of natural aging rats,and its mechanism may be related to regulating NLRP3 inflammasome,thus regulating the membrane expression of AMPA receptor,and enhancing the expression phosphorylation of CAMKⅡ and CREB.
The aim of this paper was to investigate the effect of total saponins from Panax japonicus( SPJ) on cognitive decline of natural aging rats and its mechanism. Thirty male SD rats of eighteen month old were randomly divided into three groups: aged group,10 mg·kg~(-1) SPJ-treated group and 30 mg·kg~(-1) SPJ-treated group. The SPJ-treated groups were given SPJ at the dosages of 10 mg·kg~(-1) and 30 mg·kg~(-1),respectively,from the age of 18 to 24 months. Aged group were lavaged the same amount of saline,10 six-month-old rats were used as control group,with 10 rats in each group. The open field test,novel object recognition and Morris water maze were performed to detect the changes of cognitive function in each group. The changes of synaptic transmission of long-term potentiation( LTP) in hippocampal CA1 region were detected by field potential recording. Western blot was used to detect the protein levels of NLRP3,ASC,caspase-1 and the changes of Glu A1,Glu A2,CAMKⅡ,CREB and phosphorylation of CAMKⅡ,CREB in each group.The results showed that SPJ could improve the decline of cognitive function in aging rats,reduce the damage of LTP in the hippocampal CA1 region of aged rats,and decrease the expression of NLRP3,ASC,caspase-1 in aging rats. At the same time,SPJ could enhance the membrane expression of AMPA receptor( Glu A1 and Glu A2),and increase the expression of p-CAMKⅡand p-CREB in aging rats.SPJ could improve cognitive decline of natural aging rats,and its mechanism may be related to regulating NLRP3 inflammasome,thus regulating the membrane expression of AMPA receptor,and enhancing the expression phosphorylation of CAMKⅡ and CREB.
引文
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[2] Codolo G,Plotegher N,Pozzobon T,et al. Triggering of inflammasome by aggregatedα-synuclein,an inflammatory response in synucleinopathies[J]. PLoS ONE,2013,8(1):e55375.
[3]贾占红,赵晖.竹节参总皂苷对脑缺血大鼠神经细胞凋亡和即早基因表达的影响[J].中国实验方剂学杂志,2011,17(21):168.
[4]邓丽丽,王佳文,袁丁,等.竹节参总皂苷通过NLRP1和NLRP3炎症小体途径减轻衰老大鼠神经细胞凋亡的作用研究[J].中草药,2017,48(23):4941.
[5] C Tohda,N Matsumoto,K Zou,et al. Axonal and dendritic extension by protopanaxadiol-type saponins from ginseng drugs in SK-N-SH cells[J]. Jpn J Pharmacol,2002,90(3):254.
[6]宋亚男,袁丁,张长城,等.竹节参总皂苷通过调节AMPK/Sirt1/NF-κB通路抑制衰老大鼠心肌细胞凋亡的作用研究[J].中国中药杂志,2017,42(23):4656.
[7] Turnheim K. When drug therapy gets old:pharmacokinetics and pharmacodynamics in the elderly[J]. Exp Gerontol,2003,38(8):843.
[8] Gao H,Yan P,Zhang S,et al. Long-term dietary alphalinolenic acid supplement alleviates cognitive impairment correlate with activating hippocampal CREB signaling in natural aging rats[J]. Mol Neurobiol,2016,53(7):4772.
[9]涂浩,周琴,张迅,等.竹节参总皂苷对棕榈酸诱导Hep G2细胞凋亡的保护作用研究[J].中国中药杂志,2018,43(2):390.
[10] Jobim P F C,Santos C E I D,Jeromel L,et al. Changes in the element concentration of the dorsal hippocampus CA1 region during memory consolidation and reconsolidation[J]. J Chem Neuroanat,2017,90:49.
[11] Gu X H,Xu L J,Liu Z Q,et al. The flavonoid baicalein rescues synaptic plasticity and memory deficits in a mouse model of Alzheimer's disease[J]. Behav Brain Res,2016,311.
[12]陈明,王举涛,高华武,等.基于自噬途径探讨半枝莲总黄酮抑制肿瘤细胞NLRP3炎症小体表达的机制研究[J].中国中药杂志,2017,42(24):4841.
[13] Fu A K,Hung K W,Yuen M Y,et al. IL-33 ameliorates Alzheimer's disease-like pathology and cognitive decline[J]. Proc Natl Acad Sci USA,2016,113(19):E2705.
[14] Murphy N,Cowley T R,Richardson J C,et al. The neuroprotective effect of a specific P2X7receptor antagonist derives from its ability to inhibit assembly of the NLRP3 inflammasome in glial cells[J]. Brain Pathol,2012,22(3):295.
[15] Yu S Y,Wu D C,Liu L,et al. Role of AMPA receptor trafficking in NMDA receptor-dependent synaptic plasticity in the rat lateral amygdala[J]. J Neurochem,2008,106(2):889.
[16] Li M X,Zheng H L,Luo Y,et al. Gene deficiency and pharmacological inhibition of caspase-1 confers resilience to chronic social defeat stress via regulating the stability of surface AMPARs[J]. Mol Psychiatry,2018,23(3):556.
[17] Tully T,Bourtchouladze R,Scott R,et al. Targeting the CREB pathway for memory enhancers[J]. Nat Rev Drug Discov,2003,2(4):267.
[18] Scott B R. Cyclic AMP response element-binding protein(CREB)phosphorylation:a mechanistic marker in the development of memory enhancing Alzheimer's disease therapeutics[J].Biochem Pharmacol,2012,83(6):705.
[19] Villers A,Giese K P,Ris L. Long-term potentiation can be induced in the CA1 region of hippocampus in the absence ofαCaMKⅡT286-autophosphorylation[J]. Learn Mem,2014,21(11):616.