Journal of Traditional Chinese Medical Sciences (Apr 2023)
Hupo powder promotes autophagy of menstrual blood-derived stem cells from patients with endometriosis
Abstract
Objective: To explore the effect of Hupo powder (HP) on autophagy in menstrual blood-derived stem cells (MenSCs) with endometriosis (EMT). Methods: EMT MenSCs (E-MenSCs) and healthy MenSCs (H-MenSCs) were isolated from the menstrual blood of patients with EMT and healthy female participants, respectively. We identified their stem cells’ characteristics via adipogenic and osteogenic differentiation. Twelve male Sprague–Dawley rats received 0.9% NaCl and HP-dispensing granules by gastric irrigation to prepare blank serum and medicated serum, respectively. We used serum concentrations of 5%, 10%, and 20%, each at administered times of 12, 24, and 48 h to select the best condition. These cells were divided into three groups: blank serum of the control group, blank serum of the model group, and medicated serum of the HP group. H-MenSCs were used in the control group, while E-MenSCs were used in the model and HP groups. We analyzed cell viability using a cell counting kit-8 assay, observed cell morphology, evaluated the amounts of autophagosomes and autolysosomes by transmission electron microscopy, and detected the protein expression of autophagy markers (LC3-II and Beclin1) by Western blot. Results: E-MenSCs and H-MenSCs became long fusiform with a diffuse radial pattern, forming lipid droplets and calcium nodules after adipogenic and osteogenic differentiation. We then used the best condition—20% serum and 48 h—for the subsequent experiments. In contrast to the model group, the HP group exhibited lower cell viability (P = .007), larger amounts of autophagosomes and autolysosomes (P < .001 and P = .001, respectively), and higher expression of LC3-II and Beclin1 (P = .021 and P = .019, respectively). Conclusion: Hupo powder can promote autophagy in E-MenSCs, which might be one of the mechanisms underlying its therapeutic effects.
