对药酸枣仁-合欢花对抑郁模型大鼠海马CA3区神经元凋亡GRP78蛋白表达的影响
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摘要
目的研究对药酸枣仁-合欢花对抑郁模型大鼠海马CA3区神经元凋亡及葡萄糖调节蛋白78(GRP78)蛋白表达的影响,从内质网应激(ERS)角度探讨其抗抑郁作用机制。方法将雄性SD大鼠按随机数字表法分为正常对照组、模型组、对药酸枣仁-合欢花组、盐酸文拉法辛组;采用孤养加慢性不可预见性温和应激(CUMS)建立抑郁症大鼠模型,并用糖水消耗实验观察各组大鼠的行为学变化,用苏木素-伊红(HE)染色法观察海马CA3区神经细胞形态及数量。应用DNA原位末端缺口标记(TUNEL)法检测海马CA3区神经元凋亡情况,应用免疫组化法检测海马CA3区ERS通路的关键因子GRP78蛋白表达。结果与正常对照组比较,模型组大鼠糖水偏爱值下降(P <0.01),海马CA3区神经细胞数量减少(P <0.01)、凋亡细胞数和细胞凋亡率增加(P <0.01)、GRP78蛋白表达增加(P <0.01);与模型组比较,对药酸枣仁-合欢花组、盐酸文拉法辛组大鼠糖水偏爱值升高(P <0.05),海马CA3区神经细胞数量增加(P <0.05)、凋亡细胞数和细胞凋亡率降低(P <0.05或P <0.01)、GRP78蛋白表达增加(P <0.05)。结论对药酸枣仁-合欢花能显著改善抑郁模型大鼠的抑郁行为,具有抗抑郁功效,其机制可能与提高海马CA3区GRP78蛋白表达,抑制胱天蛋白酶-12(Caspase-12)表达,减缓ERS引起的细胞凋亡,维持内质网(ER)的稳定有关。
Objective To study the effect of herbal pair spinosae Ziziphi semen and Albiziae flos on the neuronal apoptosis and the expression of GRP78 protein in the hippocampal CA3 area of the depression model rats,and to explore the antidepressant mechanism from the aspect of endoplasmic reticulum stress.Methods Male SD rats were divided into normal control group,model group,herbal pair spinosae Ziziphi semen and Albiziae flos group,and venlafaxine hydrochloride group by random number table.The depressive model rats was established by social isolation and chronic mild unpredictable stress(CUMS),and the behavioral changes of rats in each group were measured by sugar water consumption test;the morphology and quantity of neurons in the hippocampus CA3 region were observed after HE staining.Neuronal apoptosis in hippocampal CA3 region was detected by TUNEL,and the expression of GRP78 protein in the hippocampus CA3 region was detected by immunohistochemistry.Results Compared with the normal control group,the sugar water preference and number of neurons in the hippocampal CA3region decreased(P<0.01);the number of apoptotic cells and the apoptosis rate increased(P<0.01),and the expression of GRP78 protein increased(P<0.01)in the model group.Compared with the model group,the sugar water preference and number of neurons in the hippocampal CA3 region increased(P<0.05);and the number of apoptotic cells and the apoptosis rate decreased(P<0.05,P<0.01),the expression of GRP78 protein increased(P<0.05)in the herbal pair spinosae Ziziphi semen and Albiziae flos group and the venlafaxine hydrochloride group.Conclusion The herbal pair spinosae Ziziphi semen and Albiziae flos can significantly improve the symptoms of depressive model rats,and have antidepressant effect.The mechanism may be related to improving the expression of GRP78 protein in the CA3 region of the hippocampus,inhibiting Caspase-12 expression,slowing apoptosis caused by ERS,and maintaining the stability of ER.
Objective To study the effect of herbal pair spinosae Ziziphi semen and Albiziae flos on the neuronal apoptosis and the expression of GRP78 protein in the hippocampal CA3 area of the depression model rats,and to explore the antidepressant mechanism from the aspect of endoplasmic reticulum stress.Methods Male SD rats were divided into normal control group,model group,herbal pair spinosae Ziziphi semen and Albiziae flos group,and venlafaxine hydrochloride group by random number table.The depressive model rats was established by social isolation and chronic mild unpredictable stress(CUMS),and the behavioral changes of rats in each group were measured by sugar water consumption test;the morphology and quantity of neurons in the hippocampus CA3 region were observed after HE staining.Neuronal apoptosis in hippocampal CA3 region was detected by TUNEL,and the expression of GRP78 protein in the hippocampus CA3 region was detected by immunohistochemistry.Results Compared with the normal control group,the sugar water preference and number of neurons in the hippocampal CA3region decreased(P<0.01);the number of apoptotic cells and the apoptosis rate increased(P<0.01),and the expression of GRP78 protein increased(P<0.01)in the model group.Compared with the model group,the sugar water preference and number of neurons in the hippocampal CA3 region increased(P<0.05);and the number of apoptotic cells and the apoptosis rate decreased(P<0.05,P<0.01),the expression of GRP78 protein increased(P<0.05)in the herbal pair spinosae Ziziphi semen and Albiziae flos group and the venlafaxine hydrochloride group.Conclusion The herbal pair spinosae Ziziphi semen and Albiziae flos can significantly improve the symptoms of depressive model rats,and have antidepressant effect.The mechanism may be related to improving the expression of GRP78 protein in the CA3 region of the hippocampus,inhibiting Caspase-12 expression,slowing apoptosis caused by ERS,and maintaining the stability of ER.
引文
[1]梁姗,吴晓丽,胡旭,等.抑郁症研究的发展和趋势--从菌-肠-脑轴看抑郁症[J].科学通报,2018,63(20):2010-2025.
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[11]施学丽.“心肝同治”治疗抑郁症的理论探讨[J].辽宁中医杂志,2017,44(1):63-65.
[12]施学丽,郭超峰,蒋春雷,等.对药酸枣仁-合欢花对抑郁患者认知功能及血清BDNF水平的影响[J].世界科学技术-中医药现代化,2013,15(7):1604-1607.
[13]施学丽,郭超峰,范丽丽,等.对药酸枣仁-合欢花对老年抑郁症患者生活质量的影响[J].世界科学技术-中医药现代化,2013,15(9):1933-1936.
[14]施学丽,杜晓娜,郭超峰,等.不同剂量对药酸枣仁-合欢花对抑郁模型大鼠体重及行为学的影响[J].广西医学,2018,40(22):2702-2705.
[15]陈理,王玲玲.“通督调神”针法对抑郁症大鼠海马Bax和Bcl-2的影响[J].中医药学报,2011,39(6):85-88.
[16]张莹,孙黎光.GRP78的研究进展[J].国外医学(生理、病理科学与临床分册),2005(3):251-253.
[17]RAO R V,PEEL A,LOGVINOVA A,et al.Coupling endoplasmic reticulum stress to the cell death program.An Apaf-l-independent intrinsic pathway[J].J Biol Chem,2002,277(24):21836-21842.
[18]SZEGEZDI E,LOGUE S E,UORMAN A M,et al.Mediators o Iendoplasmic reticulum stress-induced apoptosis[J].EMBO Rep2006,7(9):880-885.
[19]UZNETSOV G,CHEN LB,NIGAM SK.Multiple molecular chaperonescomplex with misfolded large oligomeric glycoproteins in the endoplasmic reticulum[J].Biol Chem,1997,272(5):3057-3063.
[20]HITOMI J,KATAYAMA T,TANIGUCHI M,et al.Apoptosis induced by endoplasmic reticulum stress depends on activation of caspase-3 via caspase-12[J].Neurosci Lett,2003,357:127-130.
[2]李鹏.舒郁胶囊联合氟西汀治疗抑郁症(轻、中度)肝气郁证疗效观察及对HPA轴影响机制研究[D].济南:山东中医药大学,2016.
[3]樊蔚虹,姚建平,杨清,等.柴胡疏肝散干预卒中后抑郁模型大鼠海马神经细胞凋亡的机制研究[J].中国实验方剂学杂志,2011,17(22):216.
[4]吕莹,张德禄,张宏,等.细胞凋亡检测方法研究进展[J].西北师范大学学报:自然科学版,2007,43(2):82-87.
[5]张悦.内质网应激介导的CRH诱导大鼠海马神经元凋亡[D].石家庄:河北医科大学,2012.
[6]TABAS I,RON D.Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress[J].Nat Cell Biol,2011,13(3):184-190.
[7]GULOW K,BIENERt D,HAAS I G.BiP is feed-back regulated by control of protein translation efficiency[J].J Cell Sci,2002,115(Pt 11):2443-2452.
[8]蒋艳红,王瑾,胡梦兰,等.抑郁症易感性差异与海马CA3区及DG内树突棘数量变化的相关性研究[J].重庆医科大学学报,2018,43(6):856-862.
[9]贾宝辉,李志刚,卢峻,等.电针抗抑郁研究的模型探讨[J].针刺研究,2005,30(1):22.
[10]罗文舒,皮敏,饶晓丹,等.针刺督脉和膀胱经促进抑郁症大鼠海马神经元再生的实验研究[J].广西中医药,2008,31(4):46.
[11]施学丽.“心肝同治”治疗抑郁症的理论探讨[J].辽宁中医杂志,2017,44(1):63-65.
[12]施学丽,郭超峰,蒋春雷,等.对药酸枣仁-合欢花对抑郁患者认知功能及血清BDNF水平的影响[J].世界科学技术-中医药现代化,2013,15(7):1604-1607.
[13]施学丽,郭超峰,范丽丽,等.对药酸枣仁-合欢花对老年抑郁症患者生活质量的影响[J].世界科学技术-中医药现代化,2013,15(9):1933-1936.
[14]施学丽,杜晓娜,郭超峰,等.不同剂量对药酸枣仁-合欢花对抑郁模型大鼠体重及行为学的影响[J].广西医学,2018,40(22):2702-2705.
[15]陈理,王玲玲.“通督调神”针法对抑郁症大鼠海马Bax和Bcl-2的影响[J].中医药学报,2011,39(6):85-88.
[16]张莹,孙黎光.GRP78的研究进展[J].国外医学(生理、病理科学与临床分册),2005(3):251-253.
[17]RAO R V,PEEL A,LOGVINOVA A,et al.Coupling endoplasmic reticulum stress to the cell death program.An Apaf-l-independent intrinsic pathway[J].J Biol Chem,2002,277(24):21836-21842.
[18]SZEGEZDI E,LOGUE S E,UORMAN A M,et al.Mediators o Iendoplasmic reticulum stress-induced apoptosis[J].EMBO Rep2006,7(9):880-885.
[19]UZNETSOV G,CHEN LB,NIGAM SK.Multiple molecular chaperonescomplex with misfolded large oligomeric glycoproteins in the endoplasmic reticulum[J].Biol Chem,1997,272(5):3057-3063.
[20]HITOMI J,KATAYAMA T,TANIGUCHI M,et al.Apoptosis induced by endoplasmic reticulum stress depends on activation of caspase-3 via caspase-12[J].Neurosci Lett,2003,357:127-130.