主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJIGN UNIVERSITY OF TRADITIONAL CHINE ›› 2018, Vol. 41 ›› Issue (9): 752-758.doi: 10.3969/j.issn.1006-2157.2018.09.009

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Regulating effect of Zuogui Jiangtang Jieyu Fang on hippocampal NVU: a study based on diabetes complicated by depression*

Liu Jian1, Wang Yuhong2#, Lin Jia2, Liu Lin1, Meng Pan2, Yang Hui1, Zhao Hongqing2   

  1. 1 First Affiliated Hospital of Hunan University of Chinese Medicine, Hunan 410007, China;
    2 State Key Laboratory Base of Powdered and Innovative Chinese Medicinal of Hunan Province and Ministry of Education, Hunan 410208, China
  • Received:2018-04-04 Online:2018-09-30 Published:2018-09-30
  • Supported by:
    National Natural Science Foundation of China (No. 81573965、81603604), Natural Science Foundation of Hunan Province (No. 2017JJ3241), Education Department Scientific Research Program of Hunan Province

Abstract: Objective To discuss the influence of glutamic acid (Glu) imbalance on protein expressions of GluR2 and Beclin-1 in hippocampal neurovascular unit (NVU) in status of diabetes complicated by depression (DD), and interventional effect of Zuogui Jiangtang Jieyu Fang (Zuogui Blood Sugar-decreasing Depression-relieving Decoction, ZJJF). Methods The hippocampal neurons, astrocytes (Ast) and brain microvascular endothelial cells were primitively isolated, purified and cultured in SD rats to establish in vitro co-cultural system of hippocampal NVU. The immunocytochemistic staining method was used to identify cells, and high glucose (150 mmol/L) combined with corticosterone (CORT, 200 μmol/L) were used to intervene establishment of hippocampal NVU model. The non-modeled cells were randomly divided into normal group and blank group, and modeled cells were divided into model group, group of medicated serum of Zuogui Jiangtang Jieyu Fang (ZJJF group) and group of medicated serum of positive drugs [positive drug group, fluoxetine (0.18 g/kg)+metform (1.8 mg/kg)]. The normal group and model group were given the same amount of nutrient solution, and other groups were given medicated serum or blank serum in corresponding volume fraction of 10%. Glu content in supernatant of hippocampal NVU neurons was detected by using enzyme-linked immunosorbent assay (ELISA) in all groups. The protein expressions of GluR2 and Beclin-1 in hippocampal neurons were detected by using immunofluorescent staining and high content analysis (HCA) in all groups. The apoptosis of hippocampal neurons was detected by using Tunel staining in all groups. Results Compared with normal group and blank group, Glu content in supernatant of hippocampal NVU neurons increased significantly (P<0.01), GluR2 expression decreased significantly (P<0.01), Beclin-1 expression increased significantly (P<0.01), and apoptosis rate increased significantly in model group. Compared with model group, Glu content decreased significantly (P<0.01), GluR2 expression was up-regulated (P<0.01), Beclin-1 expression was down-regulated (P<0.01 or P<0.05), and apoptosis rate decreased significantly in positive drug group and ZJJF group. Conclusion ZJJF has significant regulation effect on the protein expressions of neuron GluR2 and Beclin-1 in hippocampal NVU model, which may be related to the relief of Glu imbalance. It may be an important mechanism of ZJJF inhibiting autophagy and antagonizing hippocampal neuron apoptosis of DD.

Key words: diabetes complicated by depression, neurovascular unit, glutamic acid, GluR2, Beclin-1, autophagy, Zuogui Jiangtang Jieyu Fang (Zuogui Blood Sugar-decreasing Depression-relieving Decoction), rats

CLC Number: 

  • R285.5