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Nanomicelle curcumin-induced DNA fragmentation in testicular tissue; Correlation between mitochondria dependent
Sana Moshari1, Vahid Nejati1, Gholamreza Najafi2
1Department of Biology, Faculty of Basic Science, Urmia University, Urmia, Iran.
Abstract:
Current study was done to assess possible anti-proliferative effect of nanomicelle curcumin (NMCM) against germ cells in testicular tissue. For this purpose, 24 mature male Wistar rats were divided into control and test groups. The animals in test groups received 7.5mg/kg, 15mg/kg and 30mg/kg of NMC (NO=6 rats in each group). Following 48days, the expression of Bcl-2, Bax, caspase-3, P53 and proliferating cell nuclear antigen (PCNA) were evaluated by using reverse transcription-PCR and immunohistochemistry. Histological changes, tubular differentiation index (TDI), tissue cellularity and serum level of testosterone were analyzed. Finally, the DNA laddering test was used to assess the DNA fragmentation as hallmark for apoptosis. The NMCM significantly (P<0.05) diminished the Bcl-2, p53 and PCNA and enhanced the Bax and caspase-3 mRNA levels. The NMCM significantly (P<0.05) elevated the percentage of Bax and caspase-3-positive tubules and remarkably reduced the percentage of tubules with positive reaction for Bcl-2, p53 and PCNA. The NCMN-received animals exhibited remarkable (P<0.05) reduction in cell population, TDI ratio and serum level of testosterone. Severe DNA fragmentation was observed in 30mg/kg NMCM-received group. In conclusion, the NMCM by reducing the testicular endocrine status, down-regulating Bcl-2 expression and by enhancing the Bax and caspase-3 expression initiates the intrinsic apoptosis pathway. On the other hand, inhibited expression of p53 and PCNA (at dose level of 30mg/kg) suppresses the p53 and PCNA-related hemostasis/preservative reactions. All these alterations adversely affect the spermatogenesis.
Insights
Nanomicelle curcumin (NMCM) negatively impacts male rat germ cells, reducing testosterone and impairing spermatogenesis by initiating apoptosis. NMCM treatment significantly alters key protein expressions and DNA integrity in testicular tissue.
Area of Science:
- Reproductive Biology
- Toxicology
- Nanomedicine
Background:
- Curcumin, a natural compound, has potential therapeutic applications.
- Nanomicelle formulations enhance curcumin bioavailability.
- Testicular germ cell proliferation and apoptosis are critical for male fertility.
Purpose of the Study:
- To evaluate the anti-proliferative effects of nanomicelle curcumin (NMCM) on testicular germ cells.
- To investigate the molecular mechanisms underlying NMCM's impact on spermatogenesis.
- To assess the dose-dependent toxicity of NMCM in male Wistar rats.
Main Methods:
- Administration of varying doses of NMCM (7.5, 15, 30 mg/kg) to male Wistar rats for 48 days.
- Analysis of gene and protein expression (Bcl-2, Bax, caspase-3, p53, PCNA) via RT-PCR and immunohistochemistry.
- Evaluation of histological changes, tubular differentiation index (TDI), cellularity, serum testosterone levels, and DNA fragmentation.
Main Results:
- NMCM significantly altered the expression of apoptosis-related genes (Bcl-2, Bax, caspase-3) and proliferation markers (p53, PCNA).
- NMCM treatment led to reduced germ cell population, decreased TDI, and lowered serum testosterone levels.
- Dose-dependent DNA fragmentation was observed, indicating NMCM-induced apoptosis.
Conclusions:
- NMCM induces intrinsic apoptosis in testicular germ cells by modulating Bcl-2, Bax, and caspase-3 expression.
- NMCM adversely affects spermatogenesis by reducing testicular endocrine status and inhibiting p53 and PCNA.
- The findings suggest NMCM possesses anti-proliferative effects on germ cells, impacting male reproductive health.