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王涵翰,张尚霞,王宇红,刘平安,赵洪庆,吴梦瑶,刘检,唐荣穗.左归降糖解郁方对DD大鼠海马NVU神经元mEPSC的影响[J].湖南中医药大学学报,2020,40(7):823-827[点击复制] |
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| 左归降糖解郁方对DD大鼠海马NVU神经元mEPSC的影响 |
| 王涵翰,张尚霞,王宇红,刘平安,赵洪庆,吴梦瑶,刘检,唐荣穗 |
| (湖南中医药大学湖南省中药粉体与创新药物省部国家重点实验室培育基地, 湖南 长沙 410208;湖南中医药大学湖南省中药粉体与创新药物省部国家重点实验室培育基地, 湖南 长沙 410208;湖南中医药大学科技创新中心, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 观察左归降糖解郁方对糖尿病并发抑郁症（diabetes mellitus with depression，DD）大鼠海马神经血管单元（neurovascular unit，NVU）体外共培养体系的神经元微小兴奋性突触后电流（miniature excitatory postsynaptic currents，mEPSC）频率及幅度的影响，探讨其对海马NVU培养体系的保护机制。方法 有效构建海马NVU体外培养体系，采用免疫细胞化学染色法进行细胞鉴定；采用高糖（150 mmol/L）联合皮质酮（200 μmol/L）干预构建海马NVU细胞模型；采用全细胞膜片钳记录NVU体外共培养体系神经元mEPSC，比较各组NVU培养体系神经元mEPSC频率和电流幅度。结果 经免疫细胞化学鉴定，NVU培养体系中细胞结构完整。与正常组和空白血清组相比，模型组大鼠海马NVU培养体系神经元mEPSC的频率和幅度均显著上升（P<0.01）；与模型组相比，左归降糖解郁方血清组和阳性药血清组大鼠海马NVU体外培养体系神经元mEPSC的频率和幅度均显著下降（P<0.01）；与阳性药血清组相比，左归降糖解郁方血清组大鼠NVU细胞培养体系神经元mEPSC频率和幅度差异均无统计学意义（P>0.05）。结论 左归降糖解郁方可降低DD大鼠海马NVU体外培养体系神经元mEPSC的频率和幅度，对海马NVU培养体系的具有调控作用。 |
| 关键词: 糖尿病 抑郁症 左归降糖解郁方 微小兴奋性突出后电流 膜片钳 |
| DOI：10.3969/j.issn.1674-070X.2020.07.009 |
| 投稿时间：2019-11-04 |
| 基金项目:国家“重大新药创制”科技重大专项资助项目（2017ZX09309026）；国家自然科学基金项目（81874464，81703694）；中医学一流学科开放基金（2018ZYX50）；湖南省研究生创新项目（CX20190566）；湖南省湖南中医药大学研究生创新项目（2018CX70）；湖南中医药大学中医学国内一流学科资助项目；湖南中医药大学研究生创新课题（2018CX68）。 |
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| Effects of Zuogui Jiangtang Jieyu Recipe on mEPSC of Hippocampal NVU Neurons in DD Rats |
| WANG Hanhan,ZHANG Shangxia,WANG Yuhong,LIU Ping'an,ZHAO Hongqing,WU Mengyao,LIU Jian,TANG Rongsui |
| (The State Key Laboratory Breeding Base for the Cooperation of Chinese Medicine Powders and Innovative Drugs of Human University of Chinese Medicine, Changsha, Hunan 410208, China;The State Key Laboratory Breeding Base for the Cooperation of Chinese Medicine Powders and Innovative Drugs of Human University of Chinese Medicine, Changsha, Hunan 410208, China;Technology Innovation Center of Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Affiliated Hospital of Human University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To observe the effects of Zuogui Jiangtang Jieyu Recipe on the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSC) in the hippocampal neurovascular unit (NVU) co-culture system in rats with diabetes mellitus with depression (DD), and to explore its protection mechanism for hippocampal NVU culture system. Methods The hippocampal NVU in vitro co-culture system was constructed efficiently. Cell identification was performed by immunocytochemical staining. Hippocampus NVU cell model was constructed by high glucose (150 mmol/L) combined with corticosterone (200 μmol/L); The whole cell patch clamp was used to record the mEPSC of NVU in vitro co-culture system, and the mEPSC frequency and current amplitude of neurons in NVU culture system were compared. Results The cell structure of NVU culture system was complete by NSE immunocytochemistry. Compared with the normal group and blank serum group, the frequency and amplitude of mEPSC in the hippocampal NVU in vitro co-culture system of the model group were significantly increased (P<0.01). Compared with the model group, the frequency and amplitude of mEPSC in the hippocampal NVU co-culture system of the Zuogui Jiangtang Jieyu Recipe serum group and the positive drug serum group were significantly decreased (P<0.01). Compared with the positive drug serum group, there was no significant difference in the frequency and amplitude of mEPSC in the neurons of the NVU cell culture system in the Zuogui Jiangtang Jieyu Recipe serum group (P>0.05). Conclusion Zuogui Jiangtang Jieyu Recipe can reduce the frequency and amplitude of neuronal mEPSC in hippocampal NVU culture system of rats with DD, and has a regulation effect on hippocampal NVU culture system. |
| Key words: diabetes mellitus depression Zuogui Jiangtang Jieyu Recipe miniature excitatory postsynaptic currents patch clamp |
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