Department of Obstetrics and Gynecology, E-Da Hospital, I-Shou University, Kaohsiung, Taiwan.
School of Medicine, College of Medicine, I-Shou University, Kaohsiung, Taiwan.
Front Endocrinol (Lausanne). 2023 May 24;14:1158527. doi: 10.3389/fendo.2023.1158527. eCollection 2023.
Endometriosis is defined as the growth of endometrial glands and stromal cells in a heterotopic location with immune dysregulation. It usually leads to chronic pelvic pain and subfertility. Although various treatments are available, the recurrence rate remains high. Adipose tissue is an abundant source of multipotent mesenchymal adipose-derived stem cells (ADSCs). ADSCs display effects on not only tissue regeneration, but also immune regulation. Thus, the current study aims to test the effects of ADSCs on the growth of endometriosis.
ADSCs isolated from lipoaspiration-generated adipose tissue and their conditioned medium (ADSC-CM) were subjected to quality validation, including karyotyping as well as growth promotion and sterility tests for microbial contamination under Good Tissue Practice and Good Manufacturing Practice regulations. An autologous endometriosis mouse model was established by suturing endometrial tissue to peritoneal wall followed by treating with DMEM/F12 medium, ADSC-CM, ADSCs or ADSC-CM+ADSCs for 28 days. The area of endometriotic cysts and the degree of pelvic adhesion were measured. ICAM-1, VEGF and caspase 3 expression was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. Moreover, the mice were allowed to mate and deliver. The pregnancy outcomes were recorded. The ADSC-CM was subjected to proteomics analysis with further data mining with Ingenuity Pathway Analysis (IPA).
Both ADSC-CM and ADSCs passed quality validation. ADSC-CM reduced the area of endometriotic cysts. The inhibition by ADSC-CM was obliterated by adding ADSCs. The presence of ADSCs with or without ADSC-CM increased the peritoneal adhesion. ADSC-CM inhibited ICAM-1 and VEGF mRNA and protein expression, whereas the addition of ADSCs not only did not inhibit by itself, but also blocked the inhibition by ADSC-CM. The resorption rate was reduced by ADSC-CM. The number of live birth/dam and the survival rate of pup at 1 week-old were both increased by ADSC-CM in mice with endometriosis. IPA demonstrated that PTX3 was potentially critical for the inhibition of endometriosis by ADSC-CM due to its anti-inflammatory and antiangiogenic properties as well as its importance in implantation.
ADSC-CM inhibited endometriosis development and improved pregnancy outcomes in mice. Potential translation to clinical treatment for human endometriosis is expected.
子宫内膜异位症是指子宫内膜腺体和间质在异位位置的生长，伴有免疫失调。它通常导致慢性盆腔疼痛和不孕。虽然有各种治疗方法，但复发率仍然很高。脂肪组织是多能间充质脂肪来源的干细胞（ADSCs）的丰富来源。ADSCs 不仅对组织再生有影响，而且对免疫调节也有影响。因此，本研究旨在测试 ADSCs 对子宫内膜异位症生长的影响。
从脂肪抽吸生成的脂肪组织中分离出 ADSCs，并对其进行条件培养基（ADSC-CM）进行质量验证，包括染色体核型分析以及根据良好组织实践和良好生产规范进行的微生物污染生长促进和无菌试验。通过将子宫内膜组织缝合到腹膜壁上来建立自体子宫内膜异位症小鼠模型，然后用 DMEM/F12 培养基、ADSC-CM、ADSCs 或 ADSC-CM+ADSCs 处理 28 天。测量子宫内膜异位症囊肿的面积和盆腔粘连的程度。通过定量逆转录聚合酶链反应（qRT-PCR）和免疫组织化学评估 ICAM-1、VEGF 和 caspase 3 的表达。此外，让小鼠交配和分娩。记录妊娠结果。对 ADSC-CM 进行蛋白质组学分析，并使用 IPA（Ingenuity Pathway Analysis）进行进一步的数据挖掘。
ADSC-CM 和 ADSCs 均通过质量验证。ADSC-CM 减少了子宫内膜异位症囊肿的面积。加入 ADSCs 消除了 ADSC-CM 的抑制作用。存在 ADSCs 或不存在 ADSC-CM 均增加了腹膜粘连。ADSC-CM 抑制了 ICAM-1 和 VEGF mRNA 和蛋白表达，而加入 ADSCs 不仅本身没有抑制作用，而且还阻断了 ADSC-CM 的抑制作用。ADSC-CM 降低了吸收率。ADSC-CM 增加了子宫内膜异位症小鼠的活产/母鼠数量和 1 周龄时幼仔的存活率。IPA 表明，由于其抗炎和抗血管生成特性以及在植入中的重要性，PTX3 可能是 ADSC-CM 抑制子宫内膜异位症的关键。
ADSC-CM 抑制了子宫内膜异位症的发展并改善了小鼠的妊娠结局。有望转化为人类子宫内膜异位症的临床治疗。
