Yin M, Chen K, Wu L, Jiang P, Ji Z, Zhang N, Zhou H, Han H
Brain Disease Center, First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China.
Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Nov 20;44(11):2063-2073. doi: 10.12122/j.issn.1673-4254.2024.11.02.
To investigate the therapeutic mechanism of Decoction (GDBSD) for improving reproductive disorders in male mouse models of Wilson disease (WD).
Sixty male homozygous TX mice were randomized equally into 4 groups and treated with daily gavage of saline (WD model group), penicillamine (0.09 g/kg), or GDBSD (0.2 mL/10 g), or with intraperitoneal injection of U0126 (20 mg/kg) in addition to GDBSD gavage, with 15 male DL mice as control. After 4 weeks of treatment, copper content in testicular tissue of the mice was detected, and histopathology of the testes and epididymis was examined using HE staining and electron microscopy. TUNEL staining was used to identify apoptotic cells in the testes. The protein expressions of Bcl-2, Cytc, caspase-3, ERK, and p-ERK in the testicular tissue were evaluated with Western blotting, and BrdU-positive cells were detected with immunohistochemical labeling. Sperm density, viability, malformation rate and fertility levels of male mice were studied.
Treatment with penicillamine and GDBSD obviously improved pathological changes of the testis, increased sperm density and motility, lowered sperm abnormality rate, fertility levels and increased testicular JOHNSEN score of TK mice, but the therapeutic effect of GDBSD was blocked by U0126. GDBSD treatment significantly lowered Cytc and caspase-3 expressions and increased Bcl-2 expression in the testicular tissue of TX mice ( < 0.05), while U0126 treatment significantly lowered testicular Bcl-2 expression level. No significant differences were found in total protein expression levels of ERK1/2 among the 5 groups, but p-ERK protein expression was significantly reduced in WD and U0126 groups and increased in penicillamine and GDBSD groups.
GDBSD can improve spermatogenesis and enhance fertility of male TX mice with WD possibly by activating the ERK signaling pathway to enhance proliferation and reduce apoptosis of the spermatogenic cells.
探讨肝豆汤（GDBSD）改善肝豆状核变性（WD）雄性小鼠模型生殖功能障碍的治疗机制。
将60只雄性纯合TX小鼠随机均分为4组，分别每日灌胃生理盐水（WD模型组）、青霉胺（0.09 g/kg）、GDBSD（0.2 mL/10 g），或在灌胃GDBSD的基础上腹腔注射U0126（20 mg/kg），以15只雄性DL小鼠作为对照。治疗4周后，检测小鼠睾丸组织中的铜含量，并用苏木精-伊红（HE）染色和电子显微镜检查睾丸和附睾的组织病理学。采用TUNEL染色鉴定睾丸中的凋亡细胞。用蛋白质免疫印迹法评估睾丸组织中Bcl-2、细胞色素C（Cytc）、半胱天冬酶-3（caspase-3）、细胞外信号调节激酶（ERK）和磷酸化细胞外信号调节激酶（p-ERK）的蛋白表达，并用免疫组织化学标记检测5-溴脱氧尿嘧啶核苷（BrdU）阳性细胞。研究雄性小鼠的精子密度、活力、畸形率和生育水平。
青霉胺和GDBSD治疗明显改善了睾丸的病理变化，提高了精子密度和活力，降低了精子畸形率、生育水平，并提高了TX小鼠的睾丸约翰森评分，但GDBSD的治疗效果被U0126阻断。GDBSD治疗显著降低了TX小鼠睾丸组织中Cytc和caspase-3的表达，增加了Bcl-2的表达（P<0.05），而U0126治疗显著降低了睾丸Bcl-2表达水平。5组间ERK1/2的总蛋白表达水平无显著差异，但WD组和U0126组的p-ERK蛋白表达显著降低，青霉胺组和GDBSD组的p-ERK蛋白表达增加。
GDBSD可能通过激活ERK信号通路增强生精细胞的增殖并减少其凋亡，从而改善WD雄性TX小鼠的精子发生并提高其生育能力。
