LncRNA PVT1修饰BMSCs移植对卵巢早衰大鼠卵巢功能及PI3K/AKT/FOXO3a通路的影响OA

Objective:To investigate the role of long noncoding RNA PVT1(LncRNA PVT1)in the treatment of cyclophosphamide-induced premature ovarian failure(POF)in rats using bone marrow mesen-chymal stem cells(BMSCs)and its potential mechanism.Methods:BMSCs were cultured in vitro and lenti-viral transfection was used to construct a LncRNA PVT1 overexpression strain(LV-LncRNA PVT1)and a negative control strain(LV-NC).Transfection efficiency was verified by real-time quantitative PCR(RT-qPCR).A POF rat model was established by intraperitoneal injection of cyclophosphamide,and estrous cycle changes were monitored to determine model success.The rats were divided into control group(no model estab-lished,intraperitoneal injection of an equal volume of normal saline),POF group(no intervention after POF model establishment),POF+BMSCs group(BMSCs were injected via the tail vein after POF model establish-ment),POF+PVT1 group(LV-LncRNA PVT1 lentivirus solution was injected via the tail vein after POF model establishment),and POF+PVT1-BMSCs group(BMSCs transfected with LV-LncRNA PVT1 lentivirus were injected via the tail vein after POF model establishment).After 4 weeks treatment,serum estradiol(E2)and follicle-stimulating hormone(FSH)levels were measured by enzyme-linked immunosorbent assay(ELISA),ovarian tissue morphology and follicle development were observed by hematoxylin and eosin(HE)staining,ovarian cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL),the expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(AKT),phosphorylated AKT(p-AKT),and forkhead transcription factor O3a(FOXO3a)proteins in ovarian tissue were measured by Western blot.Results:The expression of LncRNA PVT1 in BMSCs transfected with LV-LncRNA PVT1 was higher than that in BMSCs transfected with LV-NC lentivirus(P<0.05).Compared with the control group,the estrous cycle of the rats was disrupted after mod-eling,in addition,the serum E2 level of rats in the POF group was decreased,the FSH level was increased,the ovarian tissue atrophy occurred,the number of follicles decreased significantly,the number of atretic folli-cles increased,the TUNEL-positive cell rate was increased,the p-PI3K/PI3K and p-AKT/AKT ratios were decreased,and the expression of FOXO3a protein was downregulated(P<0.05).Compared with the POF group,the serum E2 level of rats in the POF+BMSCs,POF+PVT1,and POF+PVT1-BMSCs groups was in-creased,the FSH level was decreased,the ovarian tissue damage was alleviated,the number of follicles in-creased,the number of atretic follicles decreased,the TUNEL-positive cell rate was decreased,and the p-PI3K/PI3K,p-AKT/AKT levels,as well as the expression of FOXO3a protein were upregulated(P<0.05).The effects of the POF+PVT1-BMSCs group were stronger than those of the POF+BMSCs group(P<0.05).Conclusion:LncRNA PVT1 can enhance the repair effect of BMSCs on ovarian tissue in POF rats,reduce o-varian damage and cell apoptosis,and thus improve ovarian function.This effect may be related to the regula-tion of the PI3K/AKT/FOXO3a signaling pathway.