Human placental mesenchymal stem cells ameliorate chemotherapy-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy.
Human placental mesenchymal stem cells ameliorate chemotherapy-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy.
复制标题
人胎盘间充质干细胞通过减少细胞凋亡/氧化应激和促进自噬来改善化疗引起的睾丸损伤
DOI：
10.1186/s13287-021-02275-z
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发表时间：
2021-03-20
影响因子：
7.5
通讯作者：
Li H
中科院分区：
文献类型：
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作者：
Lu J;Liu Z;Shu M;Zhang L;Xia W;Tang L;Li J;Huang B;Li H
丁硫丹对男性生殖的副作用严重，因此在接受丁硫丹治疗的儿童中保持生育能力是全世界关注的主要问题。人胎盘间充质干细胞（hPMSCs）具有增殖稳定、免疫原性低等优点，是刺激组织修复尤其是恢复精子发生的理想材料。hPMSCs在busulfan诱导的Sertoli细胞和busulfan处理的小鼠睾丸中的保护作用尚未确定。我们的研究旨在阐明hPMSCs对布苏芬处理的睾丸和支持细胞的保护作用及其潜在机制。方法首先，建立小鼠体内布苏芬诱导的睾丸毒性模型和体外布苏芬处理的小鼠Sertoli细胞系，研究hPMSC对精子发生的保护作用及其机制。然后用游标卡尺测量睾丸的长度、宽度和重量。在移植hPMSCs后1周和4周，用苏木精-伊红染色睾丸组织切片，计数充满液体腔的精小管。通过ELISA分析，检测睾酮水平及与ROS相关的MDA、SOD、LDH、CAT活性。流式细胞仪检测ROS、细胞增殖（Ki67）和细胞凋亡（Annexin V）。接下来，用荧光显微镜检测增殖标记物（BrdU和SCP3）、抗氧化标记物（SIRT1）、精子发生标记物（PLZF）和自噬相关基因（P62和LC3AB）的荧光强度。采用qRT-PCR检测γ-H2AX、BRCA1、PARP1、PCNA、Ki67、P62、LC3 mRNA的表达。结果在巴苏凡诱导的睾丸毒性小鼠模型中，pmscs恢复了被破坏的精子发生，促进了精液参数的改善，增加了睾丸激素水平、睾丸大小和自噬。hPMSCs在体外抑制支持细胞凋亡，提高其增殖速度。此外，hPMSCs在丁硫凡诱导的睾丸毒性小鼠模型中具有抗氧化应激和降低氧化损伤的作用。此外，hPMSCs增加了增殖基因（PCNA和KI67）的表达，降低了凋亡基因γ-H2AX、BRCA1和PARP1的mRNA水平。结论本研究表明，注射hPMSC可通过减少细胞凋亡/氧化应激和促进自噬来改善丁硫芬诱导的睾丸损伤。本研究为化疗诱导精子发生的临床治疗提供了一种新的思路。
BackgroundThe side effects of busulfan on male reproduction are serious, so fertility preservation in children undergoing busulfan treatment is a major worldwide concern. Human placental mesenchymal stem cells (hPMSCs) have advantages such as stable proliferation and lower immunogenicity that make them an ideal material for stimulating tissue repair, especially restoring spermatogenesis. The protective effects of hPMSCs in busulfan-induced Sertoli cells and in busulfan-treated mouse testes have not been determined. Our study aimed to elaborate the protective effect and potential mechanisms of hPMSCs in busulfan-treated testes and Sertoli cells.MethodsFirst, we developed a mouse model of busulfan-induced testicular toxicity in vivo and a mouse Sertoli cell line treated with busulfan in vitro to assess the protective effect and mechanisms of hPMSC treatment on spermatogenesis. Then, the length, width, and weight of the testes were monitored using Vernier calipers. Furthermore, at 1 week and 4 weeks after the transplantation of hPMSCs, histological sections of testes were stained with hematoxylin-eosin, and the seminiferous tubules with fluid-filled cavities were counted. Through ELISA analysis, testosterone levels and MDA, SOD, LDH, and CAT activities, which are associated with ROS, were detected. Markers of ROS, proliferation (Ki67), and apoptosis (Annexin V) were evaluated by FACS. Next, the fluorescence intensity of proliferation markers (BrdU and SCP3), an antioxidant marker (SIRT1), a spermatogenesis marker (PLZF), and autophagy-related genes (P62 and LC3AB) were detected by fluorescence microscopy. The mRNA expression of γ-H2AX, BRCA1, PARP1, PCNA, Ki67, P62, and LC3 was determined by qRT-PCR.ResultshPMSCs restored disrupted spermatogenesis, promoted improved semen parameters, and increased testosterone levels, testis size, and autophagy in the testis toxicity mouse model induced by busulfan. hPMSCs suppressed the apoptosis of Sertoli cells and enhanced their rate of proliferation in vitro. Additionally, hPMSCs protected against oxidative stress and decreased oxidative damage in the testis toxicity mouse model induced by busulfan. Furthermore, hPMSCs increased the expression of proliferation genes (PCNA and KI67) and decreased the mRNA levels of apoptotic genes such as γ-H2AX, BRCA1, and PARP1.ConclusionsThis research showed that hPMSC injection ameliorated busulfan-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy. The present study offers an idea for a new method for clinical treatment of chemotherapy-induced spermatogenesis.
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影响因子：
7.5
作者：
Xiong Y;Wang Y;Zhang J;Zhao N;Zhang H;Zhang A;Zhao D;Yu Z;Yin Y;Song L;Xiong Y;Luan X
通讯作者：
Luan X
影响因子：
2.5
作者：
Hakemi, Sepideh Ganjalikhan;Sharififar, Fariba;Eftekhar-Vaghefi, Seyed Hassan
通讯作者：
Eftekhar-Vaghefi, Seyed Hassan
影响因子：
7.5
作者：
Ding C;Zou Q;Wang F;Wu H;Chen R;Lv J;Ling M;Sun J;Wang W;Li H;Huang B
通讯作者：
Huang B
影响因子：
2.8
作者：
Liu, Feng-Jun;Dong, Wen-Ya;Zhang, Yu-Ling
通讯作者：
Zhang, Yu-Ling
影响因子：
82.9
作者：
通讯作者：
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