黄芪甲苷对间充质干细胞分泌外泌体炎性miRNAs和炎性因子的影响OA
Effects of Astragaloside Ⅳ on Exosomal Inflammatory miRNAs and Inflammatory Factors Secreted by Mesenchymal Stem Cells
目的 探讨黄芪甲苷(AS-Ⅳ)对间充质干细胞(MSCs)分泌外泌体炎性miRNAs和炎性因子的影响.方法 从健康新生儿脐带血中分离并鉴定出MSCs,随机分为实验组和对照组.实验组用300 mg/L AS-Ⅳ干预,对照组给予等体积PBS液处理;培养24 h后,采用超速离心法结合超滤法提取MSCs分泌的外泌体MSC-Exos;通过透射电子显微镜观察MSC-Exos形态,BCA法测定MSC-Exos浓度,实时荧光定量PCR法检测MSC-Exos中炎症相关miRNAs的表达,ELISA法检测IL-1β、IL-6、IL-10、M-CSF及MCP-1含量.结果 培养24 h后,电镜下MSC-Exos形态为包膜完整的囊泡状结构,实验组MSC-Exos浓度为(1.215±0.018)μg/μL,对照组MSC-Exos浓度为(0.895±0.017)μg/μL,两组比较差异具有统计学意义(P<0.01);与对照组比较,实验组中miR-21-3p、miR-125b-5p及miR-126-3p表达上升,miR-31-5p、miR-126-5p及miR-221表达下降,差异均具统计学意义(P<0.05),两组间miR-145及miR-146a-5p表达比较,差异无统计学意义(P>0.05);与对照组比较,实验组MCP-1、M-CSF分泌水平降低,IL-6、IL-10分泌水平升高,差异均具统计学意义(P<0.05),两组间IL-1β分泌水平差异无统计学意义(P>0.05).结论 黄芪甲苷通过促进MSCs分泌负载有炎症相关miRNAs外泌体的表达,抑制促炎因子释放以调控炎症反应.
Objective To explore the effect of astragaloside Ⅳ(AS-Ⅳ)on exosomal inflammatory miRNAs and inflammatory factors secreted by mesenchymal stem cells(MSCs).Methods MSCs were isolated and identified from umbilical cord blood of healthy newborns and randomly divided into experimental and control groups.The experimental group was intervened with 300 mg/L AS-Ⅳ,while the control group was treated with equal volume of PBS solution.The exosomal MSC-Exos secreted by MSCs was extracted using ultracentrifugation combined with ultrafiltration after 24 hours of culture.Transmission electron microscopy was used to observe the morphology of MSC-Exos;the concentration of MSC-Exos was measured by BCA method;the expression of inflammation-related miRNAs was detected by RT-qPCR,and the levels of IL-1β,IL-6,IL-10,M-CSF and MCP-1 was detected by ELISA.Results After 24 hours of culture,the morphology of MSC-Exos was a vesicle-like structure with intact envelope under electron microscopy.The concentration of MSC-Exos in the experimental group was(1.215±0.018)μg/μL,and the concentration of MSC-Exos in the control group was(0.895±0.017)μg/μL,with a statistically significant difference(P<0.01).Compared with the control group,the experimental group exhibited a significant upregulation of miR-21-3p,miR-125b-5p,and miR-126-3p,the expression of miR-31-5p,miR-126-5p,and miR-221 was downregulated,with a statistically significant difference(P<0.05).No statistically significant differences were observed in miR-145 and miR-146a-5p expression between the two groups(P>0.05).Compared with the control group,the experimental group secreted MCP-1 and M-CSF levels decreased,and secreted IL-6 and IL-10 levels increased,with statistically significant differences(P<0.05),while the difference in IL-1β secretion between the two groups was not statistically significant(P>0.05).Conclusion AS-Ⅳ modulates inflammatory responses by promoting MSCs-derived exosomal secretion of inflammation-related miRNAs and suppressing pro-inflammatory factor release.
李思颖;邹晓玲;贺有缘;刘向男;薛冰倩;乔绍叶;王莹;宾马赢;罗佳;熊武
湖南中医药大学,湖南 长沙 410208||湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学,湖南 长沙 410208||湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学,湖南 长沙 410208||湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学,湖南 长沙 410208||湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学,湖南 长沙 410208||湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学,湖南 长沙 410208||湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学第一附属医院,湖南 长沙 410007湖南中医药大学第一附属医院,湖南 长沙 410007
黄芪甲苷间充质干细胞外泌体miRNAs炎性因子
Astragaloside ⅣMesenchymal stem cellsExosomesmiRNAsInflammatory factors
《中医药信息》 2026 (5)
33-38,54,7
湖南省自然科学基金医卫联合项目(2025JJ80930)长沙市自然科学基金项目(kq2502258)湖南省教育厅优秀青年项目(24B0368)
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