目的 比較子宮內(nèi)膜異位癥（EMT）患者與健康女性的經(jīng)血源間充質干細胞（MenSCs）的自噬功能。方法 分別從EMT患者及健康女性的月經(jīng)血中提取EMT MenSCs（E-MenSCs）和正常MenSCs（H-MenSCs），并借助成脂、成骨誘導分化鑒定其干細胞屬性。通過CCK-8法比較E-MenSCs和H-MenSCs在12、24、48、72、96、120 h時刻于波長450 nm的吸光度（A）值，并繪制細胞增殖曲線，比較2組細胞的細胞活力；運用透射電鏡觀察E-MenSCs與H-MenSCs的細胞形態(tài)，檢測比較其中自噬體和自噬溶酶體的數(shù)量；采用Western blotting法檢測E-MenSCs和H-MenSCs中自噬標志物微管相關蛋白輕鏈3-II（LC3-II）和Beclin1的蛋白表達。結果 E-MenSCs和H-MenSCs均為長梭形，呈輻射狀擴散，且經(jīng)成脂、成骨誘導分化后均有脂滴、鈣結節(jié)形成。與H-MenSCs比較，E-MenSCs在72、96、120 h的細胞活力均顯著升高（P<0.01、0.001），自噬體和自噬溶酶體數(shù)量顯著減少（P<0.05、0.01），且LC3-Ⅱ、Beclin1蛋白表達量也顯著降低（P<0.05）。結論 與H-MenSCs比較，E-MenSCs的自噬功能減弱，這可能是EMT發(fā)病進展的潛在機制。

Objective To compare the autophagy of menstrual blood-derived mesenchymal stem cells (MenSCs) in patients with endometriosis (EMT) and healthy women. Methods EMT MenSCs (E-MenSCs) and healthy MenSCs (H-MenSCs) were isolated from the menstrual blood of patients with endometriosis and healthy volunteers, respectively. Then their stem cell properties were identified using adipogenic and osteogenic differentiation. The absorbance (A) value at 450 nm of E-MenSCs and H-MenSCs at 12, 24, 48, 72, 96, and 120 h time points were obtained using CCK-8 assay, the cell proliferation curves were plotted, and the cell viability of E-MenSCs and H-MenSCs were compared. The cell morphology of E-MenSCs and H-MenSCs was observed by transmission electron microscope, and the number of autophagosome and autophagic lysosome was detected and compared. The expression of autophagy marker proteins (LC3-II and Beclin1) in E-MenSCs and H-MenSCs were detected by Western blotting. Results E-MenSCs and H-MenSCs, long spindle-shaped, diffused radially. After adipogenic and osteogenic differentiation, the lipid droplets and calcium nodules in E-MenSCs and H-MenSCs were observed. In contrast to H-MenSCs, E-MenSCs showed increased cell viability at 72, 96, and 120 h (P < 0.01 and 0.001), decreased expressions of autophagosomes and autolysosomes (P < 0.05 and 0.01), and reduced LC3-II and Beclin1 proteins (P < 0.05). Conclusion E-MenSCs autophagy is weakened compared with H-MenSCs, which might be the underlying mechanism of EMT pathogenesis and progression.

R966

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