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Lentivirus‐Mediated bcl‐2 Gene Therapy Improves Function and Structure of Chemotherapy‐Damaged Ovaries in Wistar Rats
Author(s) -
Wang Xuefeng,
He Yuanli,
Liu Mubiao,
Fu Xiafei
Publication year - 2013
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/aji.12048
Subject(s) - intraperitoneal injection , apoptosis , ovary , endocrinology , western blot , medicine , biology , blot , lentivirus , genetic enhancement , andrology , microbiology and biotechnology , gene , immunology , virus , biochemistry , viral disease
Objective This study aimed to explore the roles and mechanisms of lentivirus‐mediated bcl‐2 gene therapy in repairing the function and structure of chemotherapy‐damaged ovaries in rats. Method of Study The lentivirus vector carrying the bcl‐2 gene ( pGC ‐ FU ‐ EGFP ‐ bcl ‐2) was constructed and condensed at a high titer. Wistar rats were divided into seven groups based on the treatment they were given: no treatment [the normal control ( NC ) group]; intraperitoneal injection of cyclophosphamide (the CTX group); bilateral ovarian injection of pGC ‐ FU ‐ EGFP ‐ bcl ‐2 (the bcl ‐2 group) or empty vector pGC ‐ FU ‐ EGFP (the enhanced green fluorescent protein ( EGFP ) group); bilateral ovarian injection of normal saline (the NS + CTX group), pGC ‐ FU ‐ EGFP (the EGFP + CTX group), or pGC ‐ FU ‐ EGFP ‐ bcl ‐2 (the bcl ‐2 + CTX group) followed by intraperitoneal injection of CTX . At 15, 30, 45, and 60 days after injection, the rats were killed, serum levels of estradiol (E 2 ) and follicle‐stimulating hormone ( FSH ) were detected by radioimmunoassay; ovarian structure and follicles were observed under a microscope, the apoptosis of granulosa cells was detected by terminal deoxynucleotidyide transferase‐mediated biotin–dUTP biotin nick‐end labeling, and the expression of Bcl ‐2 in the ovaries was detected by Western blotting. Results After the injection of pGC ‐ FU ‐ EGFP ‐ bcl ‐2, the serum level of E 2 was elevated, whereas that of FSH was dropped, follicles were increased, the CTX ‐induced apoptosis of granulosa cells was inhibited, and the expression of Bcl ‐2 was up‐regulated. Conclusion The lentivirus‐mediated bcl‐2 gene therapy can improve ovarian function and structure damaged by chemotherapy and, therefore, might be a potential method to treat CTX ‐induced pre‐mature ovarian failure.