甘草次酸对雄黄致小鼠海马突触超微结构损伤的改善作用
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摘要
目的:探讨甘草次酸(GA)对雄黄致小鼠海马突触超微结构损伤的改善作用,并阐明其相关机制。方法:60只ICR小鼠随机分为对照组[灌胃给予0.5%羧甲基纤维素钠(CMC-Na)水溶液]、雄黄组(灌胃给予雄黄混悬液1.35g·kg-1)和GA干预组(灌胃给予GA 48mg·kg-1+雄黄1.35g·kg-1)(n=20),每日1次,连续灌胃8周。采用新事物识别实验检测各组小鼠记忆能力和认知功能,测定各组小鼠海马组织中谷胱甘肽(GSH)水平,利用透射电镜观察海马CA1区突触超微结构、突触间隙宽度、突触活性带长度、突触后致密物(PSD)厚度和突触界面曲率等结构参数的变化。结果:与对照组比较,雄黄组小鼠新物体优先指数(PI)和海马组织中GSH水平明显降低(P<0.05);与雄黄组比较,GA干预组小鼠新物体PI虽有所增大,但差异无统计学意义(P>0.05),小鼠海马组织中GSH水平升高(P<0.01)。与对照组比较,雄黄组小鼠海马CA1区突触结构模糊,突触间隙宽度明显增加(P<0.01),突触活性带长度变短(P<0.01),PSD厚度明显变薄(P<0.01),突触界面曲率减小(P<0.01);与雄黄组比较,GA干预组小鼠海马CA1区突触结构较清晰、完整,突触间隙宽度变窄(P<0.05),突触活性带长度变大(P<0.05),PSD厚度增加(P<0.05);突触界面曲率虽有所增大,但差异无统计学意义(P>0.05)。结论:雄黄可致小鼠海马突触结构参数改变,导致记忆能力和认知功能降低,GA可部分改善小鼠海马突触超微结构的异常变化。
Objective:To explore the improvement of glycyrrhetinic acid(GA)on the damage of synaptic ultrastructures of hippocampus induced by realgar in the mice,and to clarify the related mechanisms.Methods:Sixty ICR mice were randomly divided into three groups with twenty mice in each group:control group(intragastrically treated with 0.5% CMC-Na),realgar group(intragatrically treated with realgar 1.35 g·kg-1),and GA intervention group(intragastrically administered with GA 48 mg·kg-1 and realgar 1.35 g·kg-1).The mice were administrated once a day for eight consecutive weeks.The cognitive and memory abilities were tested using object recognition task(ORT).The levels of glutathione(GSH)in the hippocampus in the mice in various groups were detected.The changes of the ultrastructures of synapse in hippocampal CA1 region,the width of synaptic cleft,the length of synaptic active zone,the thickness of post synaptic density(PSD)and the curvature of synaptic interface were observed by transmission electron microscope.Results:Compared with control group,the preferential index(PI)for the novel object and the level of GSH in the hippocampus tissue of the mice in realgar group were significantly decreased(P<0.05);compared with realgar group,the PI for the novel object had no obvious changes and only showed the increasing tendency(P>0.05),and the level of GSH in the hippocampus tissue of the mice in GA intervention group was significantly increased(P<0.05).Compared with control group,the synaptic structure in the hippocampal CA1 region of the mice in realgar group was fuzzy;the width of synaptic cleft was increased(P<0.01),the length of the synaptic active zone,the thickness of PSD and the curvature of synaptic interface were decreased(P <0.01).Compared with realgar group,the synaptic structure in the hippocampal CA1 region of the mice in GA intervention group was clear and complete;the width of synaptic cleft was decreased(P<0.05),the length of the synaptic active zone and the thickness of PSD were significantly increased(P<0.05);the curvature of synaptic interface had no obvious change,only showed the increasing tendency(P>0.05).Conclusion:Realgar can change the synaptic structural parameters and cause deficits in cognitive and memory abilities.GA can alleviate the abnormal ultrastructural changes in the hippocampal synapses of the mice.
Objective:To explore the improvement of glycyrrhetinic acid(GA)on the damage of synaptic ultrastructures of hippocampus induced by realgar in the mice,and to clarify the related mechanisms.Methods:Sixty ICR mice were randomly divided into three groups with twenty mice in each group:control group(intragastrically treated with 0.5% CMC-Na),realgar group(intragatrically treated with realgar 1.35 g·kg-1),and GA intervention group(intragastrically administered with GA 48 mg·kg-1 and realgar 1.35 g·kg-1).The mice were administrated once a day for eight consecutive weeks.The cognitive and memory abilities were tested using object recognition task(ORT).The levels of glutathione(GSH)in the hippocampus in the mice in various groups were detected.The changes of the ultrastructures of synapse in hippocampal CA1 region,the width of synaptic cleft,the length of synaptic active zone,the thickness of post synaptic density(PSD)and the curvature of synaptic interface were observed by transmission electron microscope.Results:Compared with control group,the preferential index(PI)for the novel object and the level of GSH in the hippocampus tissue of the mice in realgar group were significantly decreased(P<0.05);compared with realgar group,the PI for the novel object had no obvious changes and only showed the increasing tendency(P>0.05),and the level of GSH in the hippocampus tissue of the mice in GA intervention group was significantly increased(P<0.05).Compared with control group,the synaptic structure in the hippocampal CA1 region of the mice in realgar group was fuzzy;the width of synaptic cleft was increased(P<0.01),the length of the synaptic active zone,the thickness of PSD and the curvature of synaptic interface were decreased(P <0.01).Compared with realgar group,the synaptic structure in the hippocampal CA1 region of the mice in GA intervention group was clear and complete;the width of synaptic cleft was decreased(P<0.05),the length of the synaptic active zone and the thickness of PSD were significantly increased(P<0.05);the curvature of synaptic interface had no obvious change,only showed the increasing tendency(P>0.05).Conclusion:Realgar can change the synaptic structural parameters and cause deficits in cognitive and memory abilities.GA can alleviate the abnormal ultrastructural changes in the hippocampal synapses of the mice.
引文
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[13]安思训,陈晓丽,何欣,等.多烯磷脂对青年鼠海马CA3区神经元突触可塑性的影响[J].吉林大学学报:医学版,2009,35(3):419-422.
[14]张璇,蔺俊斌,王娟,等.“形神共养”对脑缺血再灌注大鼠功能恢复和突触形态结构参数的影响[J].中国康复医学杂志,2015,30(11):1105-1111.
[15]吴敏霞,曾育琦,林旭云,等.APP/PS1转基因AD鼠海马CA1区神经元突触超微结构[J].解剖学杂志,2017,40(4):437-439.
[16]吴馥梅,杜红燕,章子贵.突触界面曲率及其生理意义[J].神经解剖学杂志,1994,10(1):89-92.
[17]LIU H Y,YANG J H,LIU Q F,et al.Lanthanum chloride impairs spatial memory through ERK/MSK1 signaling pathway of hippocampus in rats[J].Neurochem Res,2014,39(12):2479-2491.
[18]ZHANG Z G,WANG X Y,NIAN W W,et al.Effects of calcium on drinking fluorosis-induced hippocampal synaptic plasticity impairment in the offspring of rats[J].Transl Neurosci,2017,8:191-200.
[19]OZTANIR M N,CIFTCI O,CETIN A,et al.The beneficial effects of 18β-glycyrrhetinic acid following oxidative and neuronal damage in brain tissue caused by global cerebral ischemia/reperfusion in a C57BL/J6mouse model[J].Neurol Sci,2014,35(8):1221-1228.
[20]WANG D,GUO T Q,WANG Z Y,et al.ERKs and mitochondria-related pathways are essential for glycyrrhizic acid-mediated neuroprotection against glutamate-induced toxicity in differentiated PC12cells[J].Braz J Med Biol Res,2014,47(9):773-779.
[2]HUO T G,LI W K,ZHANG Y H,et al.Excitotoxicity induced by realgar in the rat hippocampus:the involvement of learning memory injury,dysfunction of glutamate metabolism and NMDA receptors[J].Mol Neurobiol,2015,51(3):980-994.
[3]董菊,项馨立,王子妤,等.牛黄解毒片配伍对雄黄肝毒性的影响及其与凋亡调控的关系[J].中国实验方剂学杂志,2015,21(24):79-83.
[4]XU C L,LIANG C H,SUN W X,et al.Glycyrrhizic acid ameliorates myocardial ischemic injury by the regulation of inflammation and oxidative state[J].Drug Des Devel Ther,2018,12:1311-1319.
[5]尤丽丽,曹东慧,姜晶,等.18β-甘草次酸对K19-C2mE转基因鼠胃肿瘤的抑制作用及其机制[J].吉林大学学报:医学版,2016,42(2):221-225.
[6]姜泓,丁敬华,张颖花,等.雄黄水飞法炮制工艺探讨[J].中药材,2009,32(1):26-28.
[7]HEYSER C J,FERRIS J S.Object exploration in the developing rat:Methodological considerations[J].Dev Psychobiol,2013,55(4):373-381.
[8]ZHANG W B,LI P L,GENG Q Q,et al.Simultaneous determination of glutathione,cysteine,homocys-teine,and cysteinylglycine in biological fluids by ion-pairing highperformance liquid chromatography coupled with precolumn derivatization[J].J Agric Food Chem,2014,62(25):5845-5852.
[9]GULDNER FH,INGHAM CA.Increase in postsynaptic density material in optic target neurons of the rat suprachiasmatic nucleus after bilateral enucleation[J].Neurosci Lett,1980,17(1/2):27-31.
[10]JONES DG,DEVON RM.An ultrastructural study into the effects of pentobarbitone on synaptic organization[J].Brain Res,1978,147(1):47-63.
[11]EAVRI R,SHEPHERD J,WELSH CA,et al.Interneuron simplification and loss of structural plasticity as markers of aging-related functional decline[J].J Neurosci,2018,38(39):8421-8432.
[12]PALACIOS-FILARDO J,MELLOR JR.Neuromodulation of hippocampal long-term synaptic plasticity[J].Curr Opin Neurobiol,2018,54:37-43.
[13]安思训,陈晓丽,何欣,等.多烯磷脂对青年鼠海马CA3区神经元突触可塑性的影响[J].吉林大学学报:医学版,2009,35(3):419-422.
[14]张璇,蔺俊斌,王娟,等.“形神共养”对脑缺血再灌注大鼠功能恢复和突触形态结构参数的影响[J].中国康复医学杂志,2015,30(11):1105-1111.
[15]吴敏霞,曾育琦,林旭云,等.APP/PS1转基因AD鼠海马CA1区神经元突触超微结构[J].解剖学杂志,2017,40(4):437-439.
[16]吴馥梅,杜红燕,章子贵.突触界面曲率及其生理意义[J].神经解剖学杂志,1994,10(1):89-92.
[17]LIU H Y,YANG J H,LIU Q F,et al.Lanthanum chloride impairs spatial memory through ERK/MSK1 signaling pathway of hippocampus in rats[J].Neurochem Res,2014,39(12):2479-2491.
[18]ZHANG Z G,WANG X Y,NIAN W W,et al.Effects of calcium on drinking fluorosis-induced hippocampal synaptic plasticity impairment in the offspring of rats[J].Transl Neurosci,2017,8:191-200.
[19]OZTANIR M N,CIFTCI O,CETIN A,et al.The beneficial effects of 18β-glycyrrhetinic acid following oxidative and neuronal damage in brain tissue caused by global cerebral ischemia/reperfusion in a C57BL/J6mouse model[J].Neurol Sci,2014,35(8):1221-1228.
[20]WANG D,GUO T Q,WANG Z Y,et al.ERKs and mitochondria-related pathways are essential for glycyrrhizic acid-mediated neuroprotection against glutamate-induced toxicity in differentiated PC12cells[J].Braz J Med Biol Res,2014,47(9):773-779.