腹腔注射白消安对精原干细胞代谢特征的影响

Effects of intraperitoneal injection of busulfan on metabolic characteristics of spermatogonial stem cells

目的通过构建白消安处理的小鼠模型,探讨其对小鼠睾丸精原干细胞(SSCs)代谢的影响。方法采用8周龄C57BL/6J雄性小鼠,腹腔注射10 mg/kg白消安,于第0天、第5天和第10天取材,并通过免疫磁珠法分选出Thy1阳性细胞。随后对这些细胞进行了纯度鉴定和代谢组学分析。结果实验发现白消安处理能明显降低小鼠睾丸质量(P<0.05),并损伤睾丸的组织结构。基于主成分分析(PCA)和偏最小二乘法判别分析(PLS-DA)模型显示,处理0 d、5 d和10 d的样本组间存在显著的代谢差异。共筛选鉴定出89种差异代谢物,包括谷胱甘肽(GSH)、牛磺酸、精氨酸、必需氨基酸(EAAs)和不饱和脂肪酸(UFAs)等,其重要相关代谢途径涉及甘油磷脂代谢、精氨酸和脯氨酸代谢等。结论白消安通过影响特定的代谢途径,产生明显的生殖毒性效应,导致小鼠睾丸质量下降和组织结构损伤。代谢组学分析其潜在的生殖毒性机制可能与脂质代谢、精氨酸和脯氨酸代谢等代谢通路有关。

Objective To establish a mouse model treated with busulfan and to investigate its effects on the metabolism of spermatogonial stem cells(SSCs)of mouse testis.Methods C57BL/6J male mice with age of 8 weeks were injected with 10 mg/kg of busulfan intraperitoneally,then Thy1 positive cells were selected by immunomagnetic beads on day 0,day 5 and day 10 and followed by identification for purity and metabolomic analysis.Results The testis weight ratio decreased and the tissue structure of testis was damaged(P<0.05).Based on the results of principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA),there were significant metabolic differences between the sample groups treated for 0 d,5 d and 10 d.A total of 89 differential metabolites were identified including glutathione(GSH),arginine and unsaturatedfatty acids(UFAs),and their important metabolic pathways involved glycerophospholipid metabolism,arginine and proline metabolism.Conclusions Affecting the specific metabolic pathway may result in obvious reproductive toxicity and lead to decrease of testicular weight as well as tissue structure damage in mice.Metabolomic analysis showed that the potential reproductive toxicity mechanism of SSCs may be related to the metabolic pathways such as lipid metabolism,arginine and proline metabolism.