背景:目前发现miR-196b-5p在细胞增殖、迁移及抑制瘢痕增生中发挥作用,但在创面愈合过程中是否发挥作用缺乏相关研究。目的:探讨脂肪干细胞源性外泌体中miR-196b-5p对烧伤创面愈合的影响。方法:构建SD大鼠皮肤深Ⅱ度烧伤模型,随机分为4...背景:目前发现miR-196b-5p在细胞增殖、迁移及抑制瘢痕增生中发挥作用,但在创面愈合过程中是否发挥作用缺乏相关研究。目的:探讨脂肪干细胞源性外泌体中miR-196b-5p对烧伤创面愈合的影响。方法:构建SD大鼠皮肤深Ⅱ度烧伤模型,随机分为4组:空白对照组、外泌体组、agomiR-196b-5p组和外泌体+antagomiR-196b-5p组,每组10只,根据不同分组于创周注射PBS、脂肪干细胞源性外泌体、miR-196b-5p激动剂和miR-196b-5p抑制剂,伤后即刻和伤后7,14,21 d观察创面愈合情况,伤后7 d苏木精-伊红染色观察创面炎症表达,伤后14 d Masson染色观察创面胶原表达及免疫组化染色观察创面CD31表达,伤后7 d Western blot检测创面中α-平滑肌肌动蛋白、Ⅰ型胶原蛋白表达。结果与结论:①agomiR-196b-5p组创面愈合较快,空白对照组和外泌体+antagomiR-196b-5p组愈合较慢;②与空白对照组和外泌体+antagomiR-196b-5p组相比,外泌体组和agomiR-196b-5p组创面炎性细胞浸润较少,CD31表达明显增加(P<0.01);③与空白对照组和外泌体+antagomiR-196b-5p组相比,外泌体组和agomiR-196b-5p组中α-平滑肌肌动蛋白、Ⅰ型胶原蛋白表达升高(P<0.05)。结果表明,脂肪干细胞源性外泌体中miR-196b-5p能促进大鼠烧伤创面愈合。展开更多
Long noncoding RNA and microRNA are regulatory noncoding RNAs that are implicated in Alzheimer's disease, but the role of long noncoding RNA-associated competitive endogenous RNA has not been fully elucidated. The...Long noncoding RNA and microRNA are regulatory noncoding RNAs that are implicated in Alzheimer's disease, but the role of long noncoding RNA-associated competitive endogenous RNA has not been fully elucidated. The long noncoding RNA growth arrest-specific 5(GAS5) is a member of the 5′-terminal oligopyrimidine gene family that may be involved in neurological disorders, but its role in Alzheimer's disease remains unclear. This study aimed to investigate the function of GAS5 and construct a GAS5-associated competitive endogenous RNA network comprising potential targets. RNA sequencing results showed that GAS5 was upregulated in five familial Alzheimer's disease(5×FAD) mice, APPswe/PSEN1dE9(APP/PS1) mice, Alzheimer's disease-related APPswe cells, and serum from patients with Alzheimer's disease. Functional experiments with targeted overexpression and silencing demonstrated that GAS5 played a role in cognitive dysfunction and multiple Alzheimer's disease-associated pathologies, including tau hyperphosphorylation, amyloid-beta accumulation, and neuronal apoptosis. Mechanistic studies indicated that GAS5 acted as an endogenous sponge by competing for microRNA-23b-3p(miR-23b-3p) binding to regulate its targets glycogen synthase kinase 3beta(GSK-3β) and phosphatase and tensin homologue deleted on chromosome 10(PTEN) expression in an Argonaute 2-induced RNA silencing complex(RISC)-dependent manner. GAS5 inhibited miR-23b-3p-mediated GSK-3β and PTEN cascades with a feedforward PTEN/protein kinase B(Akt)/GSK-3β linkage. Furthermore, recovery of GAS5/miR-23b-3p/GSK-3β/PTEN pathways relieved Alzheimer's disease-like symptoms in vivo, indicated by the amelioration of spatial cognition, neuronal degeneration, amyloid-beta load, and tau phosphorylation. Together, these findings suggest that GAS5 promotes Alzheimer's disease pathogenesis. This study establishes the functional convergence of the GAS5/miR-23b-3p/GSK-3β/PTEN pathway on multiple pathologies, suggesting a candidate therapeutic target in Alzheimer's disease.展开更多
文摘背景:目前发现miR-196b-5p在细胞增殖、迁移及抑制瘢痕增生中发挥作用,但在创面愈合过程中是否发挥作用缺乏相关研究。目的:探讨脂肪干细胞源性外泌体中miR-196b-5p对烧伤创面愈合的影响。方法:构建SD大鼠皮肤深Ⅱ度烧伤模型,随机分为4组:空白对照组、外泌体组、agomiR-196b-5p组和外泌体+antagomiR-196b-5p组,每组10只,根据不同分组于创周注射PBS、脂肪干细胞源性外泌体、miR-196b-5p激动剂和miR-196b-5p抑制剂,伤后即刻和伤后7,14,21 d观察创面愈合情况,伤后7 d苏木精-伊红染色观察创面炎症表达,伤后14 d Masson染色观察创面胶原表达及免疫组化染色观察创面CD31表达,伤后7 d Western blot检测创面中α-平滑肌肌动蛋白、Ⅰ型胶原蛋白表达。结果与结论:①agomiR-196b-5p组创面愈合较快,空白对照组和外泌体+antagomiR-196b-5p组愈合较慢;②与空白对照组和外泌体+antagomiR-196b-5p组相比,外泌体组和agomiR-196b-5p组创面炎性细胞浸润较少,CD31表达明显增加(P<0.01);③与空白对照组和外泌体+antagomiR-196b-5p组相比,外泌体组和agomiR-196b-5p组中α-平滑肌肌动蛋白、Ⅰ型胶原蛋白表达升高(P<0.05)。结果表明,脂肪干细胞源性外泌体中miR-196b-5p能促进大鼠烧伤创面愈合。
基金supported by the National Natural Science Foundation of China,Nos. 82173806 and U1803281Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Science,Nos. 2021-I2M-1-030 and 2022-I2M-2-002Non-Profit Central Research Institute Fund of Chinese Academy of Medical Sciences,No. 2022-JKCS-08 (all to RL)。
文摘Long noncoding RNA and microRNA are regulatory noncoding RNAs that are implicated in Alzheimer's disease, but the role of long noncoding RNA-associated competitive endogenous RNA has not been fully elucidated. The long noncoding RNA growth arrest-specific 5(GAS5) is a member of the 5′-terminal oligopyrimidine gene family that may be involved in neurological disorders, but its role in Alzheimer's disease remains unclear. This study aimed to investigate the function of GAS5 and construct a GAS5-associated competitive endogenous RNA network comprising potential targets. RNA sequencing results showed that GAS5 was upregulated in five familial Alzheimer's disease(5×FAD) mice, APPswe/PSEN1dE9(APP/PS1) mice, Alzheimer's disease-related APPswe cells, and serum from patients with Alzheimer's disease. Functional experiments with targeted overexpression and silencing demonstrated that GAS5 played a role in cognitive dysfunction and multiple Alzheimer's disease-associated pathologies, including tau hyperphosphorylation, amyloid-beta accumulation, and neuronal apoptosis. Mechanistic studies indicated that GAS5 acted as an endogenous sponge by competing for microRNA-23b-3p(miR-23b-3p) binding to regulate its targets glycogen synthase kinase 3beta(GSK-3β) and phosphatase and tensin homologue deleted on chromosome 10(PTEN) expression in an Argonaute 2-induced RNA silencing complex(RISC)-dependent manner. GAS5 inhibited miR-23b-3p-mediated GSK-3β and PTEN cascades with a feedforward PTEN/protein kinase B(Akt)/GSK-3β linkage. Furthermore, recovery of GAS5/miR-23b-3p/GSK-3β/PTEN pathways relieved Alzheimer's disease-like symptoms in vivo, indicated by the amelioration of spatial cognition, neuronal degeneration, amyloid-beta load, and tau phosphorylation. Together, these findings suggest that GAS5 promotes Alzheimer's disease pathogenesis. This study establishes the functional convergence of the GAS5/miR-23b-3p/GSK-3β/PTEN pathway on multiple pathologies, suggesting a candidate therapeutic target in Alzheimer's disease.
