目的 基于CXCL12/CXCR4軸探討琥珀散對子宮內(nèi)膜異位癥（EMT）經(jīng)血源間充質(zhì)干細(xì)胞（MenSCs）自噬活性的調(diào)控作用。方法 從EMT患者和健康女性志愿者的月經(jīng)血中分離提取EMT MenSCs（E-MenSCs）和正常MenSCs（HMenSCs）。采用12只7周齡雄性SD大鼠制備空白血清和含藥血清（ig給予大鼠琥珀散9.9 g·kg^-1，臨床等效劑量）。將細(xì)胞分為3組：對照組（H-MenSCs）、模型組（E-MenSCs）和琥珀散組（E-MenSCs），對照組和模型組使用20%空白血清干預(yù)，琥珀散組使用20%含藥血清干預(yù)，均干預(yù)48 h。分別采用CCK-8法、透射電鏡、實時熒光定量PCR（qRT-PCR）和Westernblotting法檢測3組細(xì)胞的細(xì)胞活力、自噬體和自噬溶酶體表達(dá)、CXCL12和CXCR4的mRNA水平和CXCL12/CXCR4及自噬標(biāo)志分子（Beclin1和LC3Ⅱ）的蛋白表達(dá)量。結(jié)果 與對照組比較，模型組的細(xì)胞活力、CXCL12和CXCR4的mRNA和蛋白表達(dá)水平均顯著提高（P<0.05、0.01、0.001），自噬體和自噬溶酶體數(shù)量、Beclin1和LC3Ⅱ的蛋白表達(dá)均顯著降低（P<0.05、0.01）；與模型組比較，琥珀散組的細(xì)胞活力、CXCL12和CXCR4的mRNA及蛋白表達(dá)水平均顯著下調(diào)（P<0.05、0.01、0.001），自噬體和自噬溶酶體數(shù)量、Beclin1和LC3Ⅱ的蛋白表達(dá)均顯著上調(diào)（P<0.05、0.01、0.001）。結(jié)論 琥珀散含藥血清能夠上調(diào)E-MenSCs自噬活性，其作用機制可能與其抑制CXCL12/CXCR4軸有關(guān)。

Objective Based on CXCL12/CXCR4 axis, the regulation of Hupo Powder on the autophagy of menstrual blood-derived mesenchymal stem cells (MenSCs) with endometriosis (EMT) was studied. Methods EMT MenSCs (E-MenSCs) and healthy MenSCs (H-MenSCs) from the menstrual blood of patients with EMT and healthy volunteers were isolated, respectively. Twelve seven-weeks old male SD rats were used to prepare blank serum and drug containing serum (9.9 g·kg^-1 of amber powder was given to rats by ig, clinical equivalent dose). These cells were divided into three groups:control (H-MenSCs), model (E-MenSCs), and Hupo Powder (E-MenSCs) groups. The control and model groups were intervened in 20% concentration of blank serum for 48 hours, while the Hupo Powder group was intervened in 20% concentration of Hupo Powder containing serum for 48 hours. CCK-8 assay, transmission electron microscope, qRT-PCR, and Western blotting were used to detect the cell viability, autophagosomes and autolysosomes, the mRNA levels of CXCL12 and CXCR4, and the protein level of CXCL12, CXCR4, and autophagy marker proteins (Beclin1 and LC3Ⅱ), respectively. Results Compared to the control group, cell viability, mRNA and protein expression of CXCL12 and CXCR4 in the model group were increased (P < 0.05, 0.01, and 0.001), and autophagosomes and autolysosomes, Beclin1 and LC3Ⅱ protein expression were decreased (P < 0.05 and 0.01). Compared to the model group, cell viability, mRNA and protein expression of CXCL12 and CXCR4 in the Hupo Powder group were down-regulated (P < 0.05, 0.01, and 0.001), and autophagosomes and autolysosomes, Beclin1 and LC3 Ⅱ protein expression were upregulated (P < 0.05, 0.01, and 0.001). Conclusion Hupo Powder containing serum can up-regulate E-MenSCs autophagy, which might relate to the inhibition of CXCL12/CXCR4 from Hupo Powder.

R285.5

國家自然科學(xué)基金面上項目（81973895）；北京中醫(yī)藥大學(xué)重點攻關(guān)項目（2020-JYB-ZDGG-143-3）