Protective Role of Coenzyme Q10 in Modulating Apoptosis, Necrosis, and Autophagy Pathways Induced by Cyclophosphamide in Rat TesticularTissue

Abstract

Cyclophosphamide (CPC) is one of the oldest anticancer drugs frequently used in cancer treatment. However, CPC is known to have adverse effects on the testis, spermatogenesis, and reproductive hormones. This study aimed to investigate the effect of Coenzyme Q10 (CoQ10), a compound naturally found in the testis and seminal fluid and used in infertility treatment, on repairing the damage caused by CPC in testicular tissue. Thirty male Wistar Albino rats were divided into five groups, each consisting of six animals: Control, CPC, CoQ10, CPC + CoQ10-I (CQ-I), and CPC + CoQ10-II (CQ-II). The mRNA expression levels of junctional genes were detected in apoptotic (caspase-8, caspase-9, caspase-3, BAX, BCL2, and BAK1), autophagic (ATG3, ATG5, and LC3), and necrotic (RIPK1) cell death mechanisms. cDNA was synthesized from total mRNA, and the gene expression levels of apoptosis markers were determined by RT-qPCR. Statistical significance was set at p<0.05. CPC was found to upregulate caspase-9, caspase-3, and BAK1 mRNA levels compared to the control group (p<0.05). However, co-administration of CoQ10 with CPC significantly reduced the expression levels of caspase-9, caspase-3, and BAK1 (p<0.05). Additionally, a significant increase in BCL-2 gene expression was observed (p<0.05). CoQ10 treatment reduced the BAX/BCL-2 ratio compared to the control and CPC groups (p<0.05). CPC reduced ATG3 and LC3 mRNA expression, but co-administration of CPC with CoQ10 increased LC3 expression (p<0.05); additionally, RIPK1 levels were lower in the CQ-I group compared to CPC (p<0.05). In conclusion, CoQ10 was found to reduce CPC-induced apoptosis, regulate autophagy, and prevent necroptosis, promoting cell survival. However, larger and longer-term studies are required for clinical implementation.

Keywords

• Cyclophosphamide
• Coenzyme Q10
• Caspase-3
• Caspase-9
• ATG3
• LC3

Sıçan Testis Dokusunda Siklofosfamid ile Tetiklenen Apoptoz, Nekroz ve Otofaji Yollarının Modülasyonunda Koenzim Q10'un Koruyucu Rolü

Abstract

Siklofosfamid (CPC) kanser tedavisinde sıklıkla kullanılan en eski antikanser ilaçlarından biridir. CPC’nin testis, spermatogenez, üreme hormonları üzerinde olumsuz etkileri olduğu bilinmektedir. Bu çalışma, infertilite tedavisinde kullanılan ve testis ile seminal sıvıda doğal olarak bulunan Koenzim Q10’un (CoQ10), CPC'nin testis dokusunda oluşturduğu hasarın onarılmasına etkisini belirlemeyi amaçlamıştır. 30 Albino Wistar erkek sıçan, her grupta 6 hayvan olacak şekilde 5 gruba [Kontrol, Siklofosfamid (CPC), Koenzim Q10 (CoQ10), CPC + CoQ10-I (CQ-I), CPC + CoQ10-II (CQ-II)] ayrıldı. Apoptotik (kaspaz-8, kaspaz-9, kaspaz-3, BAX, BCL2, ve BAK1), otofajik (ATG3, ATG5 ve LC3) ve nekrotik (RIPK1) hücre ölüm mekanizmalarındaki kavşak genlerin mRNA ekspresyon düzeyleri belirlendi. Toplam mRNA'dan cDNA sentezlendi. mRNA gen ekspresyon seviyeleri RT-qPCR ile tespit edildi. Tüm grup çalışmaları için p<0.05 istatistiksel olarak anlamlı kabul edildi. CPC’nin kontrol grubuna göre kaspaz-9, kaspaz-3 ve BAK1 mRNA düzeylerini artırdığı tespit edilmiştir (p<0.05). CPC ile birlikte CoQ10 uygulandığında, kaspaz-9, kaspaz-3 ve BAK1 genlerinin ekspresyon seviyelerini düşürdüğü (p<0.05) tespit edilmiştir. Ayrıca, BCL-2 gen ekspresyonunda istatistiksel olarak anlamlı bir artış (p<0.05) gözlemlenmiştir. Hem kontrol grubuna hem de CPC grubuna kıyasla CoQ10 uygulamasının BAX/BCL-2 oranını istatistiksel olarak azalttığı (p<0.05) ortaya konulmuştur. CPC, ATG3 ve LC3 mRNA ekspresyonunu düşürmüş, ancak CPC ile birlikte CoQ10 uygulaması LC3 ekspresyonunu artırmıştır (p<0.05); ayrıca RIPK1 seviyesi CQ-I grubunda CPC'ye göre azalmıştır (p<0.05). Sonuç olarak, CoQ10’un, siklofosfamidin (CPC) neden olduğu apoptozu düşürdüğü, otofajiyi düzenlendiği ve nekroptozu önlemede rol oynadığı ve hücre sağkalımını desteklediği tespit edilmiştir. Ancak klinik uygulamalar için daha geniş ve uzun vadeli çalışmalar gerekmektedir.

Keywords

• Siklofosfamid
• KoenzimQ10
• Kaspaz-3
• Kaspaz-9
• ATG3
• LC3

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