Feng Xiaoling, Jiang Sha, Leung WingTing, Wang Ling, Gober Hans Jürgen, Chen Lu, Zhang Yang, Wang Ling
The First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin, China.
The Heilongjiang University of Chinese Medicine, Harbin, China.
Evid Based Complement Alternat Med. 2020 Oct 5;2020:6868470. doi: 10.1155/2020/6868470. eCollection 2020.
BuShen HuoXue decoction (BSHXD) has been used to treat patients with unexplained recurrent spontaneous abortion (URSA). However, the chemical compounds and mechanism by which BSHXD exerts its therapeutic and systemic effects to promote the proliferation of decidual stromal cells (DSCs) has not been elucidated. This work sought to elucidate the cellular and molecular mechanism of BSHXD in terms of inflammatory factors IL-17A in DSCs because of the critical roles of inflammation, apoptosis, and immunity in the development and progression of pregnancy loss. Twelve migratory chemical compounds from BSHXD extract were qualitatively analyzed by high-performance liquid chromatography (HPLC). DSCs were collected from normal early pregnancy (NEP) and URSA to determine whether BSHXD affects IL-17A/IL17RA via the PI3K/AKT pathway. Abnormal apoptosis and activated p-AKT were observed in URSA DSCs. RhIL-17 A, LY294002 (a PI3K pathway inhibitor), and BSHXD were individually or simultaneously administered in NEP DSCs, suggesting that BSHXD restored cell proliferation without excessive stimulation and IL-17A promotes proliferation via the PI3K/AKT pathway. Using the same intervention in URSA DSCs, qRT-PCR measured the upregulated mRNA levels of IL-17 A/IL-17RA, PI3K, AKT, p-AKT, PTEN, Bcl-2, and Bcl-xL and downregulated mRNA levels of BAD and ACT1 after treatment with BSHXD. We demonstrated that BSHXD affected IL-17A/IL-17R via PI3K/AKT pathway to promote the proliferative activity of DSCs in URSA. These results provide a new insight to further clarify the relationship between inflammation and apoptosis and the mechanism of imbalance in the dynamic equilibrium between Th17/Treg immune cells at the maternal-fetal interface.
补肾活血汤（BSHXD）已被用于治疗不明原因复发性自然流产（URSA）患者。然而，BSHXD发挥其治疗和全身作用以促进蜕膜基质细胞（DSCs）增殖的化学成分和机制尚未阐明。由于炎症、凋亡和免疫在流产发生和发展过程中的关键作用，本研究旨在从DSCs中的炎性因子IL-17A方面阐明BSHXD的细胞和分子机制。通过高效液相色谱（HPLC）对BSHXD提取物中的12种迁移化学成分进行了定性分析。收集正常早孕（NEP）和URSA患者的DSCs，以确定BSHXD是否通过PI3K/AKT途径影响IL-17A/IL17RA。在URSA DSCs中观察到异常凋亡和活化的p-AKT。将重组人白细胞介素-17A（RhIL-17 A）、LY294002（一种PI3K途径抑制剂）和BSHXD分别或同时作用于NEP DSCs，结果表明BSHXD可在不过度刺激的情况下恢复细胞增殖，且IL-17A通过PI3K/AKT途径促进增殖。对URSA DSCs进行相同干预后，实时定量聚合酶链反应（qRT-PCR）检测到BSHXD处理后IL-17 A/IL-17RA、PI3K、AKT、p-AKT、PTEN、Bcl-2和Bcl-xL的mRNA水平上调，BAD和ACT1的mRNA水平下调。我们证明，BSHXD通过PI3K/AKT途径影响IL-17A/IL-17R，从而促进URSA中DSCs的增殖活性。这些结果为进一步阐明炎症与凋亡之间的关系以及母胎界面Th17/Treg免疫细胞动态平衡失衡机制提供了新的见解。
