左归降糖解郁方对模拟糖尿病并发抑郁症环境下海马神经血管单元的保护作用  

Protective effect of Zuogui Jiangtang Jieyu Formula on hippocampal NVU:Under simulated DD environment

作　　者：凌佳[1,2] 罗薇絮 刘灿 刘检[4] 韩远山 王宇红[1,2] LING Jia;LUO Wei-xu;LIU Can;LIU Jian;HAN Yuan-shan;WANG Yu-hong(Institute of Innovation and Applied Research in Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Provincial Key Laboratory of Power and Innovative Drugs National Key Laboratory of Ministry Training Bases,Changsha 410208,China;The Third Hospital of Changsha,Changsha 410035,China;The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学科技创新中心,长沙410208 [2]湖南省中药粉体与创新药物省部共建国家重点实验室培育基地,长沙410208 [3]长沙市第三医院,长沙410035 [4]湖南中医药大学第一附属医院,长沙410007

出　　处：《中华中医药杂志》2021年第10期5865-5869,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81874464,No.81573965);湖南中医药大学研究生创新课题(No.2020CX05)。

摘　　要：目的:探讨左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经血管单元(NVU)的保护作用。方法:构建海马NVU共培养体系,并使用高糖联合皮质酮模拟DD环境。将NVU共培养体系随机分为正常组、空白血清组、模型组、阳性药含药血清组(氟西汀0.18g/kg+二甲双胍1.8mg/kg)和中药含药血清组(32.82g/kg)。3个血清组分别给予10%的对应自制血清,正常组和模型组给予培养液补齐液面。18h后采用ELISA法检测体系中5-HT和分泌蛋白含量,采用跨内皮电阻值(TEER)和4h渗漏试验液面差评价NVU屏障功能,Western blot法检测GFAP表达情况,流式细胞术检测神经元凋亡情况。结果:与正常组比较,模型组胞内外5-HT和分泌蛋白含量均显著降低(P<0.01,P<0.05),TEER显著降低(P<0.01),4h渗漏试验液面差显著增加(P<0.01),GFAP表达显著降低(P<0.05),NE凋亡率显著增加(P<0.01);与模型组比较,中药含药血清组的胞内外5-HT和分泌蛋白含量均显著上升(P<0.05),TEER显著升高(P<0.05),4h渗漏试验液面差显著降低(P<0.05),GFAP表达显著升高(P<0.05),NE凋亡率显著降低(P<0.01)。结论:左归降糖解郁方对模拟DD环境下海马NVU具有保护作用,可能与调控NVU分泌功能有关。Objective:To investigate the regulatory effect of Zuogui Jiangtang Jieyu Formula containing serum on hippocampal neurovascular unit(NVU)barrier function under the environment of diabetes mellitus with depression(DD).Methods:Using hippocampal neurons(NE),astrocytes(AS),and brain microvascular endothelial cells(BM)to establish hippocampal NVU in vitro co-culture system,the model of DD treated by glucose combined with corticosterone for 18 h.the NVU co-culture system was randomly divided into five groups:normal group,model group,blank serum group,positive drug(fluoxetine 0.18 g/kg+metformin 1.8 mg/kg)containing serum group and Chinese herb(32.82 g/kg)containing serum group.Except the normal group and the model group were given the same amount of culture medium,the other groups were given the corresponding volume fraction of 10%containing serum or blank serum.To evaluate NVU barrier function,the changes of TEER were evaluated by cell resistance instrument,and the surface leakage height differences were detected by using micrometer.The contents of 5-HT and secretory proteins in BM and AS were detected by ELISA.Moreover,the expressions of GFAP were evaluated by Western blot and the apoptosis rate of neuron by flow cytometry.Results:Comparing with normal group,the contents of 5-HT and secretory proteins decreased significantly(P<0.01,P<0.05),the TEER decreased significantly(P<0.01),and internal and the surface leakage height increased significantly(P<0.01)in the model group.Additionally,the expression of GFAP was down-regulated and the apoptosis of NE was obvious(P<0.05,P<0.01).Fortunately,all of the above phenomena were reversed in the ZJJR group and the positive group when comparing with the model group.Conclusion:Zuogui Jiangtang Jieyu Formula could have a repair function on hippocampal NVU barrier function in the simulated DD environment,and its mechanism is related to the regulation of NVU secretion funtion.

关 键 词：糖尿病并发抑郁症 神经血管单元 屏障结构 分泌功能 左归降糖解郁方 

分 类 号：R285[医药卫生—中药学]