琥珀散通过LncRNA MALAT1/miR-142-3p促进自噬缩小子宫内膜异位症裸鼠异位病灶的机制

Mechanism of Hupo Powder promoting autophagy and reducing ectopic lesions in endometriotic nude mice by LncRNA MALAT1/miR-142-3p

目的:观察琥珀散对子宫内膜异位症(EMT)裸鼠异位病灶的干预作用,并从LncRNA MALAT1/miR-142-3p介导自噬研究其作用机制。方法:将30只7周龄雌性免疫缺陷小鼠随机分为模型组、琥珀散组(29 g/kg)与地诺孕素组(0.001 g/kg),每组10只。从5例寒凝血瘀型EMT患者月经血中提取经血源间充质干细胞(MenSCs)以制备人MenSCs植入式EMT裸鼠模型。采用HE染色法及人类白细胞抗原α(HLAA)免疫荧光法评价模型。造模成功后连续给药21 d。实时荧光定量PCR和免疫印迹法测定各组异位病灶组织中LncRNA MALAT1、miR-142-3p及自噬标志分子(Beclin1和LC3Ⅱ)的表达情况。结果:造模后,3组注射区均有囊泡样组织形成,病理切片见子宫内膜腺体和间质,免疫荧光示HLAA阳性,系造模成功。与模型组比较,琥珀散组异位病灶体积显著缩小(P<0.01),LncRNA MALAT1和Beclin1的RNA水平、Beclin1和LC3Ⅱ蛋白水平均显著升高(P<0.01,P<0.05),miR-142-3p被显著抑制(P<0.01)。结论:琥珀散能明显缩小EMT裸鼠异位病灶体积,其作用机制可能是其促进LncRNA MALAT1诱骗miR-142-3p以上调Beclin1进而增强异位内膜自噬。

Objective:To observe the effect of Hupo Powder on ectopic lesions of endometriosis(EMT)nude mice and explore the intervention mechanism on autophagy by LncRNA MALAT1/miR-142-3p pathway.Methods:A total of 30 seven-week female immunodeficiency mice were randomly divided into three groups,containing the model group(MG),Hupo Powder group(HG)and dienogest group(DG),10 mice in each group.Menstrual blood-derived stem cells(MenSCs)isolated from menstrual blood of 5 EMT patients with cold coagulation and blood stasis.Human MenSCs implanting EMT nude mice model was prepared.HE staining and human leukocyte antigenα(HLAA)immunofluorescence were used for modeling evaluation.Reverse transcription-quantitative realtime PCR and Western Blot were used for detecting the expression of LncRNA MALAT1,miR-142-3p,and autophagy marker(Beclin1 and LC3Ⅱ)in ectopic lesions each group.Results:Vesicular tissues were formed at the subcutaneous injection area in three groups after modeling.Endometrial glands and stroma were found in pathological sections,and HLAA was positively expressed.Compared with MG,ectopic lesions’volumes in HG decreased significantly(P<0.01),LncRNA MALAT1 and Beclin1 at RNA level,Beclin1 and LC3Ⅱat protein level in HG were increased(P<0.01,P<0.05),and miR-142-3p were inhibited in HG(P<0.01).Conclusion:Hupo Powder significantly reduces the ectopic lesions in EMT nude mice,which might relate to its promotion of LncRNA MALAT1 to decoy miR-142-3p,upregulate Beclin1,and enhance the autophagy of ectopic endometrium.