| 摘要: |
| 目的：基于弗林蛋白（fractalkine，FKN）/趋化因子 C-X3-C-基元受体1（chemokine C-X3-C-motif receptor 1，CX3CR1）信号研究左归降糖解郁方（Zuogui Jiangtang Jieyu presscription，ZGJTJY）调节糖尿病并发抑郁症（diabetes mellitus with depression，DD）“四重突触”体外模型的作用机制。方法：建立模拟 DD环境的体外细胞模型，实验设空白组（Control）、模型组（Model）、阳性药含药血清组（Positive）、复方中药含药血清组（ZGJTJY）、空白血清组（Blank）和CX3CR1抑制组（CX3CR1）。采用激光共聚焦检测胶质纤维酸性蛋白（glial fibrillary acidic protein，GFAP）、离子钙结合适配分子 1（ionized calcium binding adapter molecule 1，Iba-1）的表达，高内涵检测突触素（synaptophysin，SYN）、突触后致密物95（postsynaptic density protein 95，PSD-95）、FKN、CX3CR1、N-甲基-D-天冬氨酸受体2A（N-methyl-D-aspartate receptor 2A，NR2A）、N-甲基-D-天冬氨酸受体2B（N-methyl-D-aspartate receptor 2B，NR2B）的表达，酶联免疫吸附测定（enzyme-linked immunosorbent assay，ELISA）检测肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）和白细胞介素-6（interleukin-6，IL-6）等炎症因子的表达。结果：与Control比较，Model的GFAP和Iba-1荧光强度明显升高，SYN和PSD-95蛋白表达强度减弱，FKN、CX3CR1、NR2A和NR2B荧光蛋白表达量增加，TNF-α、IL-1β和IL-6含量均显著上升（P＜0.05或P＜0.01）。ZGJTJY干预后，GFAP和Iba-1降低，SYN和PSD-95升高，FKN、CX3CR1、NR2A和NR2B表达降低，TNF-α、IL-1β和IL-6含量均显著降低（P＜0.05或P＜0.01）。CX3CR1抑制组SYN和PSD-95分泌增加，FKN、CX3CR1、NR2A和NR2B表达降低（P＜0.05或P＜0.01）。结论：ZGJTJY可通过FKN/CX3CR1信号对DD信号“四重突触” 结构体外模型产生调节作用。 |
| 关键词: 糖尿病并发抑郁症 左归降糖解郁方 FKN/CX3CR1 四重突触 体外模型 |
| DOI： |
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| Mechanism of action of Zuogui Jiangtang Jieyu prescription in regulating the in vitro model of diabetes mellitus with depression with a quadruple synapse structure：A study based on the fractalkine/chemokine C-X3-C-motif receptor 1 signaling pathway |
| LIU Zhuo,LING Jia,ZHAO Hongqing |
| (Hunan Provincial Hospital of Integrated Traditional Chinese and Western Medicine，Changsha 410006，Hunan，China;Hunan Academy of Chinese Medicine，Changsha 410013，Hunan，China;Center for Science and Technology Innovation，Hunan University of Chinese Medicine，Changsha 410208，Hunan，China) |
| Abstract: |
| Objective：To investigate the mechanism of action of Zuogui Jiangtang Jieyu prescription in regulating the in vitro model of diabetes mellitus with depression （DD) with a quadruple synapse structure based on the fractalkine （FKN)/chemokine C-X3-C-motif receptor 1 （CX3CR1) signaling pathway.Methods：An in vitro cell model was established to simulate the environment of DD,the cells were divided into control group （Control)，model group （Model)，serum containing positive drug group （Positive)，serum containing Zuogui Jiangtang Jieyu prescription group （ZGJTJY)，blank serum group （Blank)，and CX3CR1 inhibition group （CX3CR1).Confocal laser scanning was used to measure the expression levels of glial fibrillary acidic protein （GFAP)，ionized calcium binding adapter molecule 1 （Iba-1)，synaptophysin （SYN)，postsynaptic density protein 95 （PSD-95)，FKN，CX3CR1，N-methyl-D-aspartate receptor 2A （NR2A)，and N-methyl-D-aspartate receptor 2B （NR2B)，and enzymelinked immunosorbent assay （ELISA) was used to measure the expression levels of the inflammatory factors tumor necrosis factor-α （TNF-α)，interleukin-1β （IL-1β)，and interleukin-6 （IL-6).Results：Compared with the Control，the Model had significant increases in the fluorescence intensities of GFAP and Iba-1，significant reductions in the protein expression levels of SYN and PSD-95，and significant increases in the protein expression levels of FKN，CX3CR1，NR2A，and NR2B and the content of TNF-α，IL-1β，and IL-6 （P < 0.05 or P < 0.01).After ZGJTJY intervention，there were significant reductions in GFAP and Iba-1，significant increases in SYN and PSD-95，and significant reductions in the expression levels of FKN，CX3CR1，NR2A，and NR2B and the content of TNF-α，IL-1β，and IL-6 （P < 0.05 or P < 0.01).The CX3CR1 inhibition group had significant increases in the secretion of SYN and PSD-95 and significant reductions in the expression levels of FKN，CX3CR1，NR2A，and NR2B （P < 0.05 or P < 0.01).Conclusion：ZGJTJY can regulate the in vitro model of DD with a quadruple synapse structure through the FKN/CX3CR1 signaling pathway. |
| Key words: diabetes mellitus with depression Zuogui Jiangtang Jieyu prescription fractalkine/chemokine C-X3-C-motif receptor 1 signaling pathway quadruple synapse in vitro model |
