Zhang Y, Zhang Y, Shen X, Wang G, Zhu L
School of Pharmacy, Wannan Medical College, Wuhu 241002, China.
Anhui Provincial Engineering Research Center for Polysaccharide Drugs, Wuhu, 241002, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Jul 20;44(7):1315-1326. doi: 10.12122/j.issn.1673-4254.2024.07.11.
To explore the neuroprotective role of Rab10 gene in depression and the mechanism mediating its effect.
Forty-eight male SD rats were randomized into a control group and 3 chronic unpredictable mild stress (CUMS) groups (=12). The rats in the latter 3 groups were subjected to injections of normal saline, an adeno-associated viral (AAV) vector, or a Rab10-overexpressing AAV vector in the lateral ventricle after CUMS modeling. The depressive behavioral changes of the rats were assessed using behavioral tests. The TargetScan database was used to predict the miRNA interacting with Rab10 and the binding sites. The interaction between miRNA-103-3p and Rab10 was investigated using dual-luciferase and radioimmunoprecipitation (RIP) assay. The effect of corticosterone treatment on PC12 cell viability was assessed with CCK-8 assay. In corticosterone-stimulated PC12 cells, the changes in BDNF, CREB, p62, Beclin-1, Wnt3a, Gsk3β, phosphorylated (p)-Gsk3β, and β-catenin protein expressions following transfection with the Rab10-overexpressing AAV vector and a miRNA-103-3p inhibitor, alone or in combination, were analyzed using qRT-PCR and Western blotting.
Injection of Rab10-overexpressing AVV vector into the lateral ventricle significantly improved depressive behaviors of CUMS rats. The mRNA and proteins expression of Rab10 were significantly down-regulated in the hippocampus of CUMS rats and in corticosteronestimulated PC12 cells. Bioinformatics analysis and the results of double luciferase and RIP experiments confirmed the targeting relationship between miRNA-103-3p and Rab10. In PC12 cells, overexpression of Rab10 or silencing miRNA-103-3p activated the Wnt/β-catenin signaling pathway, up-regulated the expressions of BDNF, CREB and Beclin-1, and down-regulated the expression of p62 protein; silencing Rab10 obviously blocked the effect of miRNA-103-3p inhibitor.
In mouse models of depression, miRNA-103-3p activates Wnt/β-catenin signaling via targeting rab10 to improve neural plasticity and promotes neural cell autophagy.
探讨Rab10基因在抑郁症中的神经保护作用及其作用机制。
将48只雄性SD大鼠随机分为对照组和3个慢性不可预测性温和应激（CUMS）组（每组n = 12）。后3组大鼠在CUMS建模后于侧脑室注射生理盐水、腺相关病毒（AAV）载体或过表达Rab10的AAV载体。采用行为学测试评估大鼠的抑郁行为变化。利用TargetScan数据库预测与Rab10相互作用的miRNA及其结合位点。采用双荧光素酶和放射免疫沉淀（RIP）实验研究miRNA - 103 - 3p与Rab10之间的相互作用。用CCK - 8法评估皮质酮处理对PC12细胞活力的影响。在皮质酮刺激的PC12细胞中，单独或联合转染过表达Rab10的AAV载体和miRNA - 103 - 3p抑制剂后，采用qRT - PCR和蛋白质免疫印迹法分析脑源性神经营养因子（BDNF）、环磷腺苷效应元件结合蛋白（CREB）、p62、Beclin - 1、Wnt3a、糖原合成酶激酶3β（Gsk3β）、磷酸化（p）- Gsk3β和β - 连环蛋白的蛋白表达变化。
向侧脑室注射过表达Rab10的AVV载体可显著改善CUMS大鼠的抑郁行为。CUMS大鼠海马及皮质酮刺激的PC12细胞中Rab10的mRNA和蛋白表达均显著下调。生物信息学分析以及双荧光素酶和RIP实验结果证实了miRNA - 103 - 3p与Rab10之间的靶向关系。在PC12细胞中，过表达Rab10或沉默miRNA - 103 - 3p可激活Wnt/β - 连环蛋白信号通路，上调BDNF、CREB和Beclin - 1的表达，下调p62蛋白表达；沉默Rab10明显阻断了miRNA - 103 - 3p抑制剂的作用。
在抑郁症小鼠模型中，miRNA - 103 - 3p通过靶向Rab10激活Wnt/β - 连环蛋白信号通路，改善神经可塑性并促进神经细胞自噬。
