中华中医药学刊

负载连翘酯苷A的HucMSC外泌体制备及其对急性肺损伤的作用机制

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郭淑杰 ¹ , 谢淑敏 ¹ , 余海艳 ² , 陈朝华 ¹ , 常万顺 ¹ , 朱艳慧 ¹ , 吴灿 ¹ , 黄海英 ¹

中华中医药学刊 | 国家项目点击 2026,44(3): 144-150

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中华中医药学刊 |国家项目点击 2026 , 44 (3) : 144 -150

负载连翘酯苷A的HucMSC外泌体制备及其对急性肺损伤的作用机制

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郭淑杰¹, 谢淑敏¹, 余海艳², 陈朝华¹, 常万顺¹, 朱艳慧¹, 吴灿¹, 黄海英¹

作者信息

^1.河南中医药大学药学院，河南郑州 450046

^2.河南省儿童医院郑州儿童医院，河南郑州 450018

通讯作者:

黄海英（1979 -），女，河南三门峡人，教授，博士研究生导师，博士，研究方向：中药新剂型的开发与应用。E-mail：huang.haiying@126.com。

作者简介:

郭淑杰（1999 -），女，河南新乡人，硕士在读，研究方向：中药新剂型的开发与应用。

Preparation of Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells Loaded with Forsythoside A and Their Mechanism of Action in Acute Lung Injury

Shujie GUO¹, Shumin XIE¹, Haiyan YU², Zhaohua CHEN¹, Wanshun CHANG¹, Yanhui ZHU¹, Can WU¹, Haiying HUANG¹

Affiliations

^1.School of Pharmacy, Henan University of Traditional Chinese Medicine, Zhengzhou 450046, Henna, China

^2.Henan Children＇s Hospital, Zhengzhou Children＇s Hospital, Zhengzhou 450018, Henna, China

出版时间: 2026-03-10 doi: 10.13193/j.issn.1673-7717.2026.03.026

文章导航

摘要

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目的

制备负载连翘酯苷A（Forsythiaside A，FTA）的人脐带间充质干细胞（Human umbilical cord mesenchymal stem cells，HucMSC）外泌体（Exosomes，Exos），并基于鞘氨醇-1-磷酸（Sphingosine-1-phosphate，S1P）/磷脂酰肌醇3-激酶（Phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B1（Protein kinase B1，AKT1）信号通路研究其对急性肺损伤（Acute lung injury，ALI）的作用机制。

方法

提取HucMSC-Exos，制备FTA-Exos，并通过马尔文粒度分析仪、透射电子显微镜和蛋白质印迹（Western blot，WB）实验对其表征，高效液相色谱法测定FTA-Exos的载药量。通过脂多糖（Lipopolysaccharide，LPS）刺激人支气管上皮细胞（Human bronchial epithelioid cells，BEAS-2B）构建ALI细胞模型，将其分为对照组、模型组、FTA组和FTA-Exos组。激光共聚焦显微镜观察BEAS-2B对FTA-Exos的摄取效果。流式细胞术检测各组细胞中活性氧（Reactive oxygen species，ROS）水平。WB实验检测各组细胞中S1P、PI3K、AKT1蛋白表达水平。

结果

HucMSC-Exos和FTA-Exos的粒径和形态均符合外泌体的生物学特征，且阳性表达标志蛋白热休克蛋白70（Heat shock protein 70，HSP70）、肿瘤易感基因101（Tumor susceptibility gene 101，TSG101）和白细胞分化抗原81（Cluster of differentiation 81，CD81），FTA-Exos载药量为（8.11 ± 0.36）%。FTA-Exos可被BEAS-2B有效摄取，且荧光强度在4 h时显著增强。与对照组比较，ALI细胞模型组ROS水平和S1P、PI3K、AKT1蛋白表达水平均显著升高（P < 0.01）；与模型组比较，FTA组和FTA-Exos组ROS水平和S1P、PI3K、AKT1蛋白表达水平均显著降低（P < 0.05，P < 0.01）。

结论

研究成功制备了FTA-Exos，并证明其可以通过影响ROS水平及S1P/PI3K/AKT1信号通路对ALI细胞模型起到调节作用，为FTA治疗ALI方向的开发应用提供了新思路。

关键词

连翘酯苷A / 人脐带间充质干细胞外泌体 / 急性肺损伤 / 活性氧 / S1P/PI3K/AKT1信号通路

Abstract

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Objective

To prepare exosomes（Exos）derived from human umbilical cord mesenchymal stem cells（HucMSC）loaded with Forsythoside A（FTA）, and investigate their mechanism of action on acute lung injury（ALI）based on the sphingosine-1-phosphate（S1P）/phosphatidylinositol 3-kinase（PI3K）/protein kinase B1（AKT1）signaling pathway.

Method

HucMSC-Exos were extracted, FTA-Exos were prepared, and characterized through Malvern particle size analyzer, transmission electron microscope and Western blot（WB）experiments. The drug loading of FTA-Exos was determined by high-performance liquid chromatography. An ALI cell model was constructed by stimulating human bronchial epithelial cells（BEAS-2B）with lipopolysaccharide（LPS）, and the cells were divided into a control group, a model group, an FTA group and an FTA-Exos group. The uptake effect of FTA-Exos by BEAS-2B cells was observed using laser confocal microscopy. Flow cytometry was used to detect the level of reactive oxygen species（ROS）in cells from each group. WB experiments were performed to detect the expression levels of S1P, PI3K and AKT1 proteins in cells of each group.

Results

The particle size and morphology of HucMSC-Exos and FTA-Exos both conformed to the biological characteristics of exosomes, and they positively expressed the marker proteins heat shock protein 70（HSP70）, tumor susceptibility gene 101（TSG101）and cluster of differentiation 81（CD81）. The drug loading of FTA-Exos was（8.11 ± 0.36）%. FTA-Exos could be effectively taken up by BEAS-2B cells, and the fluorescence intensity was significantly enhanced at 4 hours. Compared with those in the control group, the levels of ROS and the expressions of S1P, PI3K and AKT1 proteins were significantly increased in the ALI cell model group（P < 0.01）. In comparison with those in the model group, the levels of ROS and the expressions of S1P, PI3K and AKt1 proteins were significantly decreased in both the FTA group and the FTA-Exos group（P < 0.05, P < 0.01）.

Conclusion

The study successfully prepared FTA-Exos and demonstrated that they can regulate ALI cell models by affecting ROS levels and the S1P/PI3K/AKT1 signaling pathway, providing new ideas for the development and application of FTA in the treatment of ALI.

Key words

Forsythiaside A / HucMSC-Exos / ALI / reactive oxygen species / S1P/PI3K/AKT1 signaling pathway

引用本文

郭淑杰, 谢淑敏, 余海艳, 陈朝华, 常万顺, 朱艳慧, 吴灿, 黄海英. 负载连翘酯苷A的HucMSC外泌体制备及其对急性肺损伤的作用机制. 中华中医药学刊, 2026 , 44 (3) : 144 -150 . DOI: 10.13193/j.issn.1673-7717.2026.03.026

Shujie GUO, Shumin XIE, Haiyan YU, Zhaohua CHEN, Wanshun CHANG, Yanhui ZHU, Can WU, Haiying HUANG. Preparation of Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells Loaded with Forsythoside A and Their Mechanism of Action in Acute Lung Injury[J]. Chinese Archives of Traditional Chinese Medicine, 2026 , 44 (3) : 144 -150 . DOI: 10.13193/j.issn.1673-7717.2026.03.026

基金

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国家自然科学基金青年科学基金项目(81904021; 82204736; 82404876)
河南省省级科技研发计划项目联合基金项目(222301420090)
河南省“十四五”规划重点项目(2023JKZD16)
河南中医药大学研究生科研创新能力提升计划项目(2023KYCX067)

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2026年第44卷第3期

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文章信息

doi: 10.13193/j.issn.1673-7717.2026.03.026

首发时间：2026-10-14
出版时间：2026-03-10

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基金

国家自然科学基金青年科学基金项目(81904021; 82204736; 82404876)

河南省省级科技研发计划项目联合基金项目(222301420090)

河南省“十四五”规划重点项目(2023JKZD16)

河南中医药大学研究生科研创新能力提升计划项目(2023KYCX067)

作者信息

^1.河南中医药大学药学院，河南郑州 450046

^2.河南省儿童医院郑州儿童医院，河南郑州 450018

通讯作者:

黄海英（1979 -），女，河南三门峡人，教授，博士研究生导师，博士，研究方向：中药新剂型的开发与应用。E-mail：huang.haiying@126.com。

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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