Table 2.
Results of hUCMSC-Exos treatment.
| Methods | Results | P value | |
|---|---|---|---|
| Exosomal treatment in vivo | (1) Vaginal smear | Estrous cycle was gradually restored to normal after treatment | |
| (2) Morphological analysis | Follicles were significantly increased in the Exos group (Figure 3D) | ||
| (3) Ovarian follicle counts | Numbers of primordial, primary, secondary, mature, and atretic follicles were significantly increased following exosomal transplantation (Figure 3E) | Primordial: POI vs Normal, P < 0.05; POI vs Exos, P < 0.05. Primary: POI vs Normal, P < 0.05; POI vs Exos, P < 0.05. Secondary: POI vs Normal, P < 0.05; POI vs Exos, P >0.05. Mature: POI vs Normal, P < 0.05; POI vs Exos, P < 0.01. Atretic: POI vs Normal, P < 0.01; POI vs Exos, P < 0.01. | |
| (4) Body weights | Body weights in the Exos group exhibited a gradual increase (Figure 3F) | ||
| (5) Hormones levels in serum | AMH and E2 were significantly elevated, while FSH levels were suppressed after exosomal administration (Figure 3G-I) | AMH: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P < 0.05. E2: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P < 0.05. FSH: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P > 0.05. | |
| (6) Reproductive tests | Exosomes significantly improved reproductive functions of POI mice model (Figures 4A–E) | Number of offspring: POI vs Normal, P < 0.001; POI vs Exos, P < 0.05; Exos vs Normal, P > 0.05. Time of birth: POI vs Normal, P < 0.001; POI vs Exos, P < 0.01; Exos vs Normal, P > 0.05. | |
| (7) FSHR expression in ovaries | I Immunofluorescence and Immunohistochemistry staining of FSHR revealed that more functional GCs were observed in the Exos group (Figures 5A, B) | ||
| (8) Proliferative ability | Immunofluorescence staining of PCNA and Ki67, Immunohistochemistry staining of Ki67 showed hUCMSC-Exos promoted ovarian proliferation (Figures 6A–C) | ||
| (9) Mechanism exploration | hUCMSC-Exos promoted ovarian proliferation by regulating the Hippo pathway (Figures 6D–K) | FSHR: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P < 0.01. PCNA: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P < 0.01. YAP1: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P > 0.05. p-YAP: POI vs Normal, P < 0.001; POI vs Exos, P < 0.05; Exos vs Normal, P < 0.01. TEAD1: POI vs Normal, P < 0.001; POI vs Exos, P < 0.01; Exos vs Normal, P > 0.05. TAZ: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P > 0.05. MST1: POI vs Normal, P < 0.001; POI vs Exos, P < 0.001; Exos vs Normal, P < 0.01. | |
| Exosomal treatment in vitro | (1) EdU assay | Proliferation of GCs was significantly increased in the Exos group (Figures 2C, D) | Cell proliferation ratio: CTX vs Normal, P < 0.01; CTX vs Exos, P < 0.01; Exos vs Normal, P < 0.05. |
| (2) CCK-8 assay | Proliferation of GCs was significantly enhanced in the Exos group (Figure 2E) | ||
| (3) Western blot | PCNA and FSHR were elevated in the Exos group (Figures 2F–H) | PCNA: CTX vs Normal, P < 0.01; CTX vs Exos, P < 0.01; Exos vs Normal, P > 0.05. FSHR: CTX vs Normal, P < 0.001; CTX vs Exos, P < 0.001; Exos vs Normal, P > 0.05. | |
| Therapeutic Mechanism | (1) EdU assay | hUCMSC-Exos promoted GC proliferation in vitro by regulating the Hippo pathway and the effect was inhibited by a YAP inhibitor (Figures 7A, B) | Cell proliferation ratio: CTX vs Normal, P < 0.001; CTX vs Exos, P < 0.01; Exos vs Normal, P < 0.01; Exos-Verteporfin vs Exos, P < 0.01; Exos-Verteporfin vs CTX, P >0.05. |
| (2) CCK-8 assay | hUCMSC-Exos promoted GC proliferation and the effect was inhibited by a YAP inhibitor (Figure 7C) | ||
| (3) Western blot | hUCMSC-Exos elevated GCs proliferation and function by regulating the Hippo pathway (Figures 7D–K) | FSHR: CTX vs Exos, P < 0.001; Exos vs Exos-Verteporfin, P < 0.001. PCNA: CTX vs Exos, P < 0.001; Exos vs Exos-Verteporfin, P < 0.001. YAP1: CTX vs Exos, P < 0.01; Exos vs Exos-Verteporfin, P < 0.01. p-YAP: CTX vs Exos, P < 0.01; Exos vs Exos-Verteporfin, P < 0.001. TEAD1: CTX vs Exos, P < 0.001; Exos vs Exos-Verteporfin, P < 0.001. TAZ: CTX vs Exos, P < 0.001; Exos vs Exos-Verteporfin, P < 0.001. MST1: CTX vs Exos, P < 0.001; Exos vs Exos-Verteporfin, P < 0.001. |