左归降糖解郁方对糖尿病并发抑郁症大鼠海马NVU连接蛋白表达的影响
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摘要
目的基于神经血管单元(NVU)探讨左归降糖解郁方对糖尿病并发抑郁症大鼠海马紧密和缝隙连接蛋白表达的影响。方法 70只成年雄性SD大鼠高脂乳剂灌胃、尾静脉注射链脲佐菌素联合慢性不可预见性应激建立糖尿病并发抑郁症动物模型,并随机分为5组:模型组,二甲双胍+氟西汀组,左归降糖解郁方高、中、低剂量组,另取12只未造模大鼠作为空白对照组,于应激造模同时灌胃给药28 d。旷场实验和Morris水迷宫评价大鼠自主活动水平和学习记忆能力,透射电镜观察大鼠海马NVU形态结构,免疫组化和实时荧光定量PCR检测大鼠海马紧密连接蛋白Claudin-1、Occludin和缝隙连接蛋白43(CX43)、水通道蛋白4(AQP4)的表达量。结果与空白对照组相比,模型组大鼠自主活动次数和空间探索时间明显减少、逃避潜伏期明显延长(P<0.01);透射电镜和病理结果显示模型组大鼠海马内皮细胞肿胀,血管周围紧密连接和缝隙连接紊乱,细胞排列松散,Claudin-1、Occludin、CX43和AQP4及其mRNA表达显著降低(P<0.01)。与模型组相比,左归降糖解郁方高剂量组自主活动次数和空间探索时间显著增加、逃避潜伏期显著减少(P<0.05、P<0.01);海马区域细胞水肿减轻,血管周围紧密和缝隙连接恢复,细胞排列较为整齐均一,Claudin-1、Occludin、CX43和AQP4及其mRNA表达明显升高(P<0.05、P<0.01)。结论左归降糖解郁方能明显缓解大鼠抑郁样行为,其机制可能与增强大鼠海马紧密和缝隙连接蛋白表达,维持海马NVU整体结构与功能有关。
Objective To discuss the influence of Zuogui Jiangtang Jieyu Fang(ZHJYF, Left-Restoring Sugar-lowering Depression-relieving Formula), based on neurovascular unit(NVU), on the expressions of tight junction proteins and connexins in rats with diabetes mellitus(DM) complicated by depression. Methods Adult SD rats(n=70) were orally given high-fat emulsion, injection of streptozocin(STZ) in caudal vein and chronic unpredicted stress to establish the model of DM complicated by depression. The rats were randomly divided into 5 groups: model group, metformin+fluoxetine group, high-dose, mid-dose and low-dose ZHJYF groups. Other 12 unmodeling rats were chosen into blank control group. All group were orally given corresponding medicinals at the same time of stress for 28 d. The level of autonomic activity and learning-memory ability were reviewed by using open-field test and Morris water maze(MWM), and morphologic structure of hippocampal NVU was observed by using transmission electron microscope(TEM). The expressions of tight junction proteins(Claudin-1 and Occluding), connexins [aonnexin43(CX43) and aquaporin4(AQP4)] were detected by using immunohistochemistry technique and RT-qPCR. Results Compared with blank control group, the times of autonomic activity and space exploration time decreased significantly and escape latency was prolonged significantly in model group(P<0.01). The results of TEM and pathology showed that swollen hippocampal endothelial cells, confused perivascular tight and gap junctions and loosely arranged cells, and the protein and mRNA expressions of Claudin-1, Occludin, CX43 and AQP4 decreased significantly in model group(P<0.01). Compared with mode group, the times of autonomic activity and space exploration time increased significantly and escape latency was shortened significantly(P<0.05 or P<0.01), swelling of hippocampal endothelial cells was relieved, perivascular tight and gap junctions was recovered, and cell arrangement was tidiness, and the protein and mRNA expressions of Claudin-1, Occludin, CX43 and AQP4 increased significantly in high-dose ZHJYF group(P<0.05 or P<0.01). Conclusion ZHJYF can significantly relieve depressive behavior in rats, and the mechanism may be related to enhance of expressions of tight junction proteins and connexins and maintain the structure and function of hippocampal NVU.
Objective To discuss the influence of Zuogui Jiangtang Jieyu Fang(ZHJYF, Left-Restoring Sugar-lowering Depression-relieving Formula), based on neurovascular unit(NVU), on the expressions of tight junction proteins and connexins in rats with diabetes mellitus(DM) complicated by depression. Methods Adult SD rats(n=70) were orally given high-fat emulsion, injection of streptozocin(STZ) in caudal vein and chronic unpredicted stress to establish the model of DM complicated by depression. The rats were randomly divided into 5 groups: model group, metformin+fluoxetine group, high-dose, mid-dose and low-dose ZHJYF groups. Other 12 unmodeling rats were chosen into blank control group. All group were orally given corresponding medicinals at the same time of stress for 28 d. The level of autonomic activity and learning-memory ability were reviewed by using open-field test and Morris water maze(MWM), and morphologic structure of hippocampal NVU was observed by using transmission electron microscope(TEM). The expressions of tight junction proteins(Claudin-1 and Occluding), connexins [aonnexin43(CX43) and aquaporin4(AQP4)] were detected by using immunohistochemistry technique and RT-qPCR. Results Compared with blank control group, the times of autonomic activity and space exploration time decreased significantly and escape latency was prolonged significantly in model group(P<0.01). The results of TEM and pathology showed that swollen hippocampal endothelial cells, confused perivascular tight and gap junctions and loosely arranged cells, and the protein and mRNA expressions of Claudin-1, Occludin, CX43 and AQP4 decreased significantly in model group(P<0.01). Compared with mode group, the times of autonomic activity and space exploration time increased significantly and escape latency was shortened significantly(P<0.05 or P<0.01), swelling of hippocampal endothelial cells was relieved, perivascular tight and gap junctions was recovered, and cell arrangement was tidiness, and the protein and mRNA expressions of Claudin-1, Occludin, CX43 and AQP4 increased significantly in high-dose ZHJYF group(P<0.05 or P<0.01). Conclusion ZHJYF can significantly relieve depressive behavior in rats, and the mechanism may be related to enhance of expressions of tight junction proteins and connexins and maintain the structure and function of hippocampal NVU.
引文
[1] Darwish L,Beroncal E,Sison MV,et al.Depression in people with type 2 diabetes:current perspectives[J].Diabetes,Metabolic Syndrome and Obesity:Targets and Therapy,2018,11:333-343.
[2] 徐雅岚.模拟糖尿病并发抑郁症环境下海马区神经血管单元体外模型的建立及中药干预[D].长沙:湖南中医药大学,2016.Xu YL.Establishment of in vitro model of hippocampa neurovascular unit in condition of simulated diabetes mellitus complicated by depression and intervention with Chinese medicine[D].Changsha:Hunan University of Traditional Chinese Medicine,2016.
[3] Huang SG,Guo WL,Zhou ZC,et al.Altered expression levels of occludin claudin-1 and myosin light chain kinase in the common bile duct of pediatric patients with pancreaticobiliary maljunction[J].BMC Gastroenterol,2016,16:7.
[4] Jullienne A,Fukuda AM,Ichkova A,et al.Modulating the water channel AQP4 alters miRNA expression,astrocyte connectivity and water diffusion in the rodent brain[J].Scientific Reports,2018,8:4186.
[5] 刘检,王宇红,凌佳,等.左归降糖解郁方对糖尿病并发抑郁症海马NVU神经元的调控作用及其机制[J].北京中医药大学学报,2018,41(9):752-758.Liu J,Wang YH,Ling J,et al.Regulating effect of Zuogui Jiangtang Jieyu Fang on hippocampal NVU:a study based on diabetes complicated by depression[J].Journal of Beijing University of Traditional Chinese Medicine,2018,41(9):752-758.
[6] 赵洪庆,杜青,凌佳,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元JNK/Elk-1/c-fos通路的调控作用[J].中国临床药理学与治疗学,2016,21(6):625-629.Zhao HQ,Du Q,Ling J,et al.Regulatory effects of Zuoguijiangtangjieyu formula on JNK/Elk-1 /c-fos pathway in hippocampal neurons of diabetic rats with depression[J].Chin J Clin Pharmacol Ther,2016,21(6):625-629.
[7] 李佳帅,朱路文,叶涛,等.神经血管单元与神经退行性疾病关系研究进展[J].中国康复理论与实践,2018,24(3):287-289.Li JS,Zhu LW,Ye T,et al.Relationship of neurovascular units and neurodegenerative diseases (review)[J].Chin J Rehabil Theory Prac,2018,24(3):287-289.
[8] 张颖,王联生,陈宇霞,等.基于神经血管单元稳态探索开心解郁方抗抑郁作用机制[J].世界科学技术-中医药现代化,2017,19(8):1355-1359.Zhang Y,Wang LS,Chen YX,et al.Effect of Kai-Xin Jie-Yu Decoction on regulation of neurovascular unit of depression[J].World Science and Technology/Modernization of Traditional Chinese Medicine and Materia Medica,2017,19(8):1355-1359.
[9] Aggarwal A,Khera A,Singh I,et al.S-nitrosoglutathione prevents blood-brain barrier disruption associated with increased matrix metalloproteinase-9 activity in experimental diabetes[J].Journal of Neurochemistry,2015,132(3):595-608.
[10] Falkowska A,Gutowska I,Goschorska M,et al.Energy metabolism of the brain,including the cooperation between astrocytes and neurons,especially in the context of glycogen metabolism[J].Int J Mol Sci,2015,16(11):25959-25981.
[11] 徐雅岚,赵洪庆,杜青,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马区星形胶质细胞谷氨酸转运体的影响[J].北京中医药大学学报,2016,39(6):470-475.Xu YL,Zhao HQ,Du Q,et al.Effect of Zuogui Jiangtang Jieyu Formula on amino acid transporter expression in hippocampal astrocytes of diabetes mellitus rats with depression[J].Journal of Beijing University of Traditional Chinese Medicine,2016,39(6):470- 475.
[12] 薛占霞,高永山,武海霞,等.大黄酚对原代培养小鼠星形胶质细胞谷氨酸代谢的影响[J].北京中医药大学学报,2016,39(1):35-39.Xue ZX,Gao YS,Wu HX,et al.Effects of chrysophanol on glutamate metabolism of primarily cultured strocytes in mice[J].Journal of Beijing University of Traditional Chinese Medicine,2016,39(1):35-39.
[13] 黄燕,王庆松.星形胶质细胞的水通道蛋白4在神经系统疾病相关认知障碍中的作用[J].国际神经病学神经外科学杂志,2018,45(1):99-102.Huang Y,Wang QS.Role of aquaporin 4 in astrocyte in cognitive impairment related to neurological diseases[J].Journal of International Neurology and Neurosurgery,2018,45(1):99-102.
[14] Golden SH,Shah N,Naqibuddin M,et al.The prevalence and specificity of depression diagnosis in a clinic-based population of adults with type 2 diabetes mellitus[J].Psychosomatics,2017,58(1):28-37.
[2] 徐雅岚.模拟糖尿病并发抑郁症环境下海马区神经血管单元体外模型的建立及中药干预[D].长沙:湖南中医药大学,2016.Xu YL.Establishment of in vitro model of hippocampa neurovascular unit in condition of simulated diabetes mellitus complicated by depression and intervention with Chinese medicine[D].Changsha:Hunan University of Traditional Chinese Medicine,2016.
[3] Huang SG,Guo WL,Zhou ZC,et al.Altered expression levels of occludin claudin-1 and myosin light chain kinase in the common bile duct of pediatric patients with pancreaticobiliary maljunction[J].BMC Gastroenterol,2016,16:7.
[4] Jullienne A,Fukuda AM,Ichkova A,et al.Modulating the water channel AQP4 alters miRNA expression,astrocyte connectivity and water diffusion in the rodent brain[J].Scientific Reports,2018,8:4186.
[5] 刘检,王宇红,凌佳,等.左归降糖解郁方对糖尿病并发抑郁症海马NVU神经元的调控作用及其机制[J].北京中医药大学学报,2018,41(9):752-758.Liu J,Wang YH,Ling J,et al.Regulating effect of Zuogui Jiangtang Jieyu Fang on hippocampal NVU:a study based on diabetes complicated by depression[J].Journal of Beijing University of Traditional Chinese Medicine,2018,41(9):752-758.
[6] 赵洪庆,杜青,凌佳,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马神经元JNK/Elk-1/c-fos通路的调控作用[J].中国临床药理学与治疗学,2016,21(6):625-629.Zhao HQ,Du Q,Ling J,et al.Regulatory effects of Zuoguijiangtangjieyu formula on JNK/Elk-1 /c-fos pathway in hippocampal neurons of diabetic rats with depression[J].Chin J Clin Pharmacol Ther,2016,21(6):625-629.
[7] 李佳帅,朱路文,叶涛,等.神经血管单元与神经退行性疾病关系研究进展[J].中国康复理论与实践,2018,24(3):287-289.Li JS,Zhu LW,Ye T,et al.Relationship of neurovascular units and neurodegenerative diseases (review)[J].Chin J Rehabil Theory Prac,2018,24(3):287-289.
[8] 张颖,王联生,陈宇霞,等.基于神经血管单元稳态探索开心解郁方抗抑郁作用机制[J].世界科学技术-中医药现代化,2017,19(8):1355-1359.Zhang Y,Wang LS,Chen YX,et al.Effect of Kai-Xin Jie-Yu Decoction on regulation of neurovascular unit of depression[J].World Science and Technology/Modernization of Traditional Chinese Medicine and Materia Medica,2017,19(8):1355-1359.
[9] Aggarwal A,Khera A,Singh I,et al.S-nitrosoglutathione prevents blood-brain barrier disruption associated with increased matrix metalloproteinase-9 activity in experimental diabetes[J].Journal of Neurochemistry,2015,132(3):595-608.
[10] Falkowska A,Gutowska I,Goschorska M,et al.Energy metabolism of the brain,including the cooperation between astrocytes and neurons,especially in the context of glycogen metabolism[J].Int J Mol Sci,2015,16(11):25959-25981.
[11] 徐雅岚,赵洪庆,杜青,等.左归降糖解郁方对糖尿病并发抑郁症大鼠海马区星形胶质细胞谷氨酸转运体的影响[J].北京中医药大学学报,2016,39(6):470-475.Xu YL,Zhao HQ,Du Q,et al.Effect of Zuogui Jiangtang Jieyu Formula on amino acid transporter expression in hippocampal astrocytes of diabetes mellitus rats with depression[J].Journal of Beijing University of Traditional Chinese Medicine,2016,39(6):470- 475.
[12] 薛占霞,高永山,武海霞,等.大黄酚对原代培养小鼠星形胶质细胞谷氨酸代谢的影响[J].北京中医药大学学报,2016,39(1):35-39.Xue ZX,Gao YS,Wu HX,et al.Effects of chrysophanol on glutamate metabolism of primarily cultured strocytes in mice[J].Journal of Beijing University of Traditional Chinese Medicine,2016,39(1):35-39.
[13] 黄燕,王庆松.星形胶质细胞的水通道蛋白4在神经系统疾病相关认知障碍中的作用[J].国际神经病学神经外科学杂志,2018,45(1):99-102.Huang Y,Wang QS.Role of aquaporin 4 in astrocyte in cognitive impairment related to neurological diseases[J].Journal of International Neurology and Neurosurgery,2018,45(1):99-102.
[14] Golden SH,Shah N,Naqibuddin M,et al.The prevalence and specificity of depression diagnosis in a clinic-based population of adults with type 2 diabetes mellitus[J].Psychosomatics,2017,58(1):28-37.