| 摘要: | 臨床常見治療癌症之化療藥物Cisplatin (CIS)造成睪丸功能障礙之副作用會影響男性生殖功能。睪丸內細胞因氧化壓力增加會促使發炎反應，細胞走向凋亡損傷，亦會發生精子生成異常，最終造成睪丸功能性受損。而L-半胱胺酸 (L-cysteine, CYS)因具有強大抗氧化、抗發炎等功效，但至今對男性生殖影響之相關機制研究尚未明確證實，故本研究目的為考慮臨床輔助治療之應用，以體內及體外模式探討CYS對CIS造成男性生殖損傷之相關改善效應。體外實驗利用TM3及TM4小鼠睪丸細胞株，以MTS試驗及結晶紫染色測定細胞存活率，以Western blot測定血睪障壁、發炎及細胞凋亡相關蛋白表現。結果顯示，CYS可顯著恢復CIS誘導TM3及TM4細胞存活率，並減少Caspase3、PARP、Bax凋亡相關蛋白表現，同時降低TM3細胞NLRP3及COX2發炎相關蛋白表現，且增加TM4細胞ZO-1結構蛋白表現。亦以CIS建立誘導睪丸損傷之動物模式，實驗期21天後，發現CYS可顯著降低睪丸組織及精子結構損傷，維持血清睪固酮濃度，恢復精子存活狀態，並且減少睪丸PARP蛋白表現。因此，本研究表明L-cysteine可改善Cisplatin對男性生殖之不利影響，顯示L-cysteine具有輔助臨床Cisplatin藥物治療下對男性生殖功能損傷之保護潛力。
Cisplatin (CIS), cis-diamminedichloroplatinum (II), a widely used chemotherapeutic drug in different types of cancers. Commonly, CIS can adversely affect testicular injury by inducing oxidative, inflammation, and apoptosis to lead to spermatogenesis abnormality. The non-essential amino acid L-cysteine (CYS) is used within cells for diverse roles, including antioxidant homeostasis and anti-inflammatory. Therefore, the aim of this study was to evaluate the possible protective effect of L-cysteine on cisplatin-induced male reproductive damage through in vivo and in vitro study. In in vitro study, TM3 and TM4 cells were co-treated with different concentrations of CYS (0.1, 0.2, 0.5, 1 mM) and induced by CIS (7.5, 10 ?M) for 24 and 16 hr. In in vivo study, SD rats were administered low CYS (LC, 100 mg/kg bw/day) or high CYS (HC, 300 mg/kg bw/day) for 21 consecutive days and then injected CIS (10 mg/kg bw, i.p.) single dose to establish reproductive damage model. The results show that CYS significantly recovered cisplatin-induced TM3/4 cell death by MTS assay and crystal violet staining. Besides, CYS significantly reduced apoptosis (PARP, caspase3, Bax), inflammation (NLRP3, COX2) and increased blood-testis barrier (ZO-1) related protein expressions by western blotting. In animal study, sperm analysis showed that CYS significantly alleviated sperm viability after cisplatin-induced reproductive damage. The serum level of testosterone had significantly increased by CYS. Besides, the protein expression of PARP in testis had significantly decreased. In conclusion, L-cysteine exhibited a potential protective effect on CIS-induced testicular and spermatogenic impairment through in vivo and in vitro study. |
