| 摘要: |
| 目的：研究左归降糖解郁方调控糖尿病并发抑郁症大鼠海马微小RNA-let-7d（microRNAs-let-7d，miRNA-let-7d）/TLX/微小RNA-9（microRNAs-9，miRNA-9）信号通路并促进齿状回（dentate gyrus，DG）颗粒下区（subgranular zone，SGZ）神经干细胞（neural stem cells，NSC）增殖的作用机制。方法：用尾静脉注射链脲佐菌素（streptozotocin，STZ，38 mg/kg）联合慢性不可预测性温和刺激复制糖尿病并发抑郁症大鼠模型。将造模成功的大鼠随机分为模型组、腺相关对照病毒组（阴性对照组，2 μL）、过表达TLX腺相关病毒组（过表达TLX组，2 μL）、左归降糖解郁方组+TLX敲低腺相关病毒组（ZGJJ+TLX敲低组，20.52 g/kg+2 μL）、左归降糖解郁方组（ZGJJ组，20.52 g/kg），另设正常组。采用血糖仪检测各组大鼠空腹血糖（fasting blood-glucose，FBG）水平，酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测外周胰岛素（insulin，INS）及DG区白细胞介素-4（interleukin -4，IL-4）、白细胞介素-1β（interleukin -1β,IL-1β）水平，旷场实验评价大鼠抑郁样行为，苏木素-伊红（eosin-hematoxylin，HE）染色评价大鼠海马SGZ病理变化，免疫荧光法检测SGZ胸苷类似物溴脱氧尿苷（thymidine analog bromodeoxyuridine，BrdU）蛋白的表达情况，实时荧光定量聚合酶链反应法（real-time fluorescence quantitative polymerase chain reaction，RT-qPCR）检测miRNA-let-7d/TLX/miRNA-9基因的表达。结果：与正常组比较，模型组大鼠跨格总路程明显减少，不动时间明显增加，FBG和INS水平显著升高，成熟颗粒细胞数量明显减少，病理改变明显，DG区BrdU积分光密度值减少，IL-4含量降低，IL-1β含量增加，miRNA-let-7d和microRNA-9表达增加，TLX mRNA表达减少（均P＜0.05）;与模型组比较，ZGJJ组和ZGJJ+TLX敲低组大鼠跨格总路程明显增加，不动时间明显缩短，FBG和INS水平显著降低，成熟颗粒细胞数量明显增加，DG区BrdU积分光密度值增加，IL-4分泌增加，IL-1β分泌抑制，miRNA-let-7d和miRNA-9表达减少，TLX mRNA表达增加（均P＜0.05）。与ZGJJ组比较，ZGJJ+TLX敲低组FBG和INS水平差异无统计学意义（P＞0.05），跨格总路程减少，不动时间增加，成熟颗粒细胞数量减少，DG区BrdU积分光密度值减少，IL-4含量降低，IL-1β含量增加，microRNA-let-7d和microRNA-9表达增加，TLX mRNA表达减少（均P＜0.05）。结论：左归降糖解郁方可通过调控海马microRNA-let-7d/TLX/microRNA-9信号通路促进SGZ神经干细胞增殖。 |
| 关键词: 糖尿病并发抑郁症 颗粒下区 TLX 微小RNA-let-7d 微小RNA-9 |
| DOI： |
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| Mechanism of action of Zuogui Jiangtang Jieyu prescription in regulating the microRNAs/TLX pathway and promoting the proliferation of neural stem cells in the subgranular zone of rats with diabetes complicated by depression |
| DU Qing,GUO Xuejun,LIU Zhuo |
| (Hunan Academy of Chinese Medicine，Changsha 410013，Hunan，China;Hunan Provincial Hospital of Integrated Traditional Chinese and Western Medicine，Changsha 410006，Hunan，China) |
| Abstract: |
| Objective: To investigate the mechanism of action of Zuogui Jiangtang Jieyu prescription in regulating the microRNA-let-7d (miRNA-let-7d）/TLX/microRNA-9 (miRNA-9） signaling pathway in the hippocampus and promoting the proliferation of neural stem cells (NSCs） in the subgranular zone (SGZ） of the dentate gyrus (DG） in rats with diabetes complicated by depression.Methods: Rats were given injection of 38 mg/kg streptozotocin (STZ） via the caudal vein and chronic unpredictable mild stimulation to establish a model of diabetes complicated by depression.After successful modeling，the rats were randomly divided into model group，adeno-associated virus control group (negative control group，2 μL），adeno-associated virus TLX overexpression group (TLX overexpression group，2 μL），Zuogui Jiangtang Jieyu prescription+adeno-associated virus TLX knockdown group (ZGJJ+TLX knockdown group，20.52 g/kg+2 μL），and Zuogui Jiangtang Jieyu prescription group (ZGJJ group，20.52 g/kg），and a normal group was also established.A glucometer was used to measure the level of fasting blood glucose (FBG）;ELISA was used to measure the level of insulin (INS） in peripheral blood and the levels of interleukin-4 (IL-4） and interleukin-1β (IL-1β） in the DG;the open field test was used to assess the depression-like behavior of rats;HE staining was used to observe the pathological changes of the SGZ in the hippocampus of rats;immunofluorescence assay was used to measure the protein expression of the thymidine analog bromodeoxyuridine (BrdU） in the SGZ，and RT-qPCR was used to measure the mRNA expression levels of miRNA-let-7d，TLX，and miRNA-9.Results: Compared with the normal group，the model group had a significant reduction in the total distance of grids crossed，a significant increase in immobility time，significant increases in the levels of FBG and INS，a significant reduction in the number of mature granular cells，and marked pathological changes，as well as a significant reduction in the integrated optical density of BrdU in the DG，a significant reduction in the content of IL-4，a significant increase in the content of IL-1β，significant increases in the expression levels of miRNA-let-7d and microRNA-9，and a significant reduction in the mRNA expression level of TLX (all P＜0.05）.Compared with the model group，the ZGJJ group and the ZGJJ+TLX knockdown group had a significant increase in the total distance of grids crossed，a significant reduction in immobility time，significant reductions in the levels of FBG and INS，a significant increase in the number of mature granular cells，a significant increase in the integrated optical density of BrdU in the DG，a significant increase in the secretion of IL-4，a significant inhibition of IL-1β secretion，significant reductions in the expression levels of miRNA-let-7d and microRNA-9，and a significant increase in the mRNA expression level of TLX (all P＜0.05）.There were no significant differences in the levels of FBG and INS between the ZGJJ group and the ZGJJ+TLX knockdown group (P＞0.05），and compared with the ZGJJ group，the ZGJJ+TLX knockdown group had a significant reduction in the total distance of grids crossed，a significant increase in immobility time，a significant reduction in the number of mature granular cells，a significant reduction in the integrated optical density of BrdU in the DG，a significant reduction in the content of IL-4，a significant increase in the content of IL-1β，significant increases in the expression levels of miRNA-let-7d and microRNA-9，and a significant reduction in the mRNA expression level of TLX (all P＜0.05）.Conclusion: Zuogui Jiangtang Jieyu prescription can promote the proliferation of NSCs in the SGZ by regulating the microRNA-let-7d/TLX/microRNA-9 signaling pathway in the hippocampus. |
| Key words: diabetes complicated by depression subgranular zone TLX microRNA-let-7d microRNA-9 |
