Abstract：
BACKGROUND: WuHuTang (WHT) is a traditional Chinese medicine compound for treating asthma, and the evidence supports that it has a good effect on acute asthma attacks in children and adults. Respiratory syncytial virus (RSV) is an important factor in the pathogenesis of acute asthma attacks, and the effect on dendritic cells is the key to its pathogenesis. Previous studies have confirmed that the pathogenesis of viruses is related to exosomes. However, there are few studies on the exosomes induced by RSV. Whether WHT can improve the changes caused by RSV-induced exosomes or not is worthy of further exploration.
OBJECTIVE: We aim to study the effects of RSV-induced exosomes on the function and autophagy of dendritic cells, and to observe the intervention effect of WHT serum on the above effects.
METHODS: The co-culture model of exosomes derived from bone marrow mesenchymal stem cells induced by RSV (BMSCs-Exo-RSV) and dendritic cells was established, and then WHT serum was used to intervene. After 24 h of intervention, the CCK-8 method, flow cytometry, Elisa, RT-qCPR, and Western blot were used to detect the above-mentioned culture model.
RESULTS: RSV-induced exosomes had certain effects on viability, apoptosis, and costimulatory molecules generation of dendritic cells. At the same time, the levels of IL-6, IL-12, TNF-α, and autophagy increased, while the levels of IL-4, IL-10, and TGF-β decreased, and the AKT/TSC/mTOR pathway was inhibited. WHT serum could activate this pathway and reverse the above changes in dendritic cells.
CONCLUSIONS: This study reveals that the pathogenic effect of RSV is related to the exosomes induced by RSV. The exosomes induced by RSV affect the function of dendritic cells by inhibiting the AKT/TSC/mTOR pathway, which can be activated by WHT to reverse the effects caused by RSV-induced exosomes.
OBJECTIVE: We aim to study the effects of RSV-induced exosomes on the function and autophagy of dendritic cells, and to observe the intervention effect of WHT serum on the above effects.
METHODS: The co-culture model of exosomes derived from bone marrow mesenchymal stem cells induced by RSV (BMSCs-Exo-RSV) and dendritic cells was established, and then WHT serum was used to intervene. After 24 h of intervention, the CCK-8 method, flow cytometry, Elisa, RT-qCPR, and Western blot were used to detect the above-mentioned culture model.
RESULTS: RSV-induced exosomes had certain effects on viability, apoptosis, and costimulatory molecules generation of dendritic cells. At the same time, the levels of IL-6, IL-12, TNF-α, and autophagy increased, while the levels of IL-4, IL-10, and TGF-β decreased, and the AKT/TSC/mTOR pathway was inhibited. WHT serum could activate this pathway and reverse the above changes in dendritic cells.
CONCLUSIONS: This study reveals that the pathogenic effect of RSV is related to the exosomes induced by RSV. The exosomes induced by RSV affect the function of dendritic cells by inhibiting the AKT/TSC/mTOR pathway, which can be activated by WHT to reverse the effects caused by RSV-induced exosomes.
摘要：
背景：五虎汤（WHT）是一种治疗哮喘的中药复方，证据支持它对儿童和成人急性哮喘发作有良好的疗效。呼吸道合胞病毒(RSV)是哮喘急性发作的重要发病因素,而对树突状细胞的作用是其发病机制的关键。以往的研究已经证实病毒的发病机制与外泌体有关。然而,关于RSV诱导的外泌体的研究很少。WHT能否改善RSV诱导的外泌体引起的变化值得进一步探索。
目的：我们旨在研究RSV诱导的外泌体对树突状细胞功能和自噬的影响，并观察WHT血清对上述影响的干预效果。
方法：建立RSV诱导骨髓间充质干细胞外泌体（BMSCs-Exo-RSV）与树突状细胞共培养模型，然后用WHT血清进行干预。经过24小时的干预，CCK-8方法，流式细胞术,Elisa,RT-qCPR,采用Westernblot检测上述培养模型。
结果：RSV诱导的外泌体对生存力有一定的影响，凋亡，和共刺激分子产生树突状细胞。同时,IL-6、IL-12、TNF-α、自噬增加，而IL-4，IL-10和TGF-β的水平降低，AKT/TSC/mTOR通路受到抑制。WHT血清可以激活该途径并逆转树突状细胞的上述变化。
结论：本研究提示RSV的致病作用与RSV诱导的外泌体有关。RSV诱导的外泌体通过抑制AKT/TSC/mTOR通路影响树突状细胞的功能,可以被WHT激活以逆转RSV诱导的外泌体引起的作用。
目的：我们旨在研究RSV诱导的外泌体对树突状细胞功能和自噬的影响，并观察WHT血清对上述影响的干预效果。
方法：建立RSV诱导骨髓间充质干细胞外泌体（BMSCs-Exo-RSV）与树突状细胞共培养模型，然后用WHT血清进行干预。经过24小时的干预，CCK-8方法，流式细胞术,Elisa,RT-qCPR,采用Westernblot检测上述培养模型。
结果：RSV诱导的外泌体对生存力有一定的影响，凋亡，和共刺激分子产生树突状细胞。同时,IL-6、IL-12、TNF-α、自噬增加，而IL-4，IL-10和TGF-β的水平降低，AKT/TSC/mTOR通路受到抑制。WHT血清可以激活该途径并逆转树突状细胞的上述变化。
结论：本研究提示RSV的致病作用与RSV诱导的外泌体有关。RSV诱导的外泌体通过抑制AKT/TSC/mTOR通路影响树突状细胞的功能,可以被WHT激活以逆转RSV诱导的外泌体引起的作用。
