Han Shanshan, Dai Yanlin, Sun Lihui, Xing Yaping, Ding Ying, Zhang Xia, Xu Shanshan
Pediatric Medical College, Henan University of Traditional Chinese Medicine, Zhengzhou, China.
Department of Pediatrics, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, China.
Front Pharmacol. 2023 Feb 17;14:1050907. doi: 10.3389/fphar.2023.1050907. eCollection 2023.
We determined the effects of Cuscutae semen ( Lam. or R. Br.)-Radix rehmanniae praeparata ( Libosch.) on the protein levels in testicular tissues of rats gavaged with tripterygium wilfordii multiglycosides (GTW) and elucidated the molecular mechanism underlying Cuscutae semen-Radix rehmanniae praeparata for relieving GTW-induced reproductive injury. A total of 21 male Sprague-Dawley rats were randomly divided into the control group, model group, and Cuscutae semen-Radix rehmanniae praeparata group based on their body weights. The control group was given 10 mLkg of 0.9% normal saline by gavage daily. The model group (GTW group) was administered with 12 mg kg GTW by gavage daily. Cuscutae semen-Radix rehmanniae praeparata group (the TSZSDH group) was administered with 1.56 gkg of Cuscutae semen-Radix rehmanniae praeparata granules daily according to their model group dosing. The serum levels of luteinizing hormone, follicle-stimulating hormone, estradiol, and testosterone were measured after 12 weeks of continuous gavage, and the pathological lesion of testicular tissues was observed. Differentially expressed proteins were evaluated by quantitative proteomics and verified by western blotting (WB) and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR). Cuscutae semen-Radix rehmanniae praeparata can effectively relieve pathological lesions of GTW-induced testicular tissues. A total of 216 differentially expressed proteins were identified in the TSZSDH group and model group. High-throughput proteomics revealed that differentially expressed proteins are closely associated with the peroxisome proliferator-activated receptor (PPAR) signaling pathway, protein digestion and absorption, and protein glycan pathway in cancer. Cuscutae semen-Radix rehmanniae praeparata can significantly upregulate the protein expressions of Acsl1, Plin1, Dbil5, Plin4, Col12a1, Col1a1, Col5a3, Col1a2, Dcn, so as to play a protective role on testicular tissues. Acsl1, Plin1, and PPARγ on the PPAR signaling pathway were verified by WB and RT-qPCR experiments, which were found to be consistent with the results of proteomics analysis. Cuscutae semen and Radix rehmanniae praeparata may regulate the PPAR signaling pathway mediated Acsl1, Plin1 and PPARγ to reduce the testicular tissue damage of male rats caused by GTW.
我们研究了菟丝子（Lam. 或R. Br.）-熟地黄（Libosch.）对雷公藤多苷（GTW）灌胃大鼠睾丸组织蛋白质水平的影响，并阐明了菟丝子-熟地黄减轻GTW诱导的生殖损伤的分子机制。将21只雄性Sprague-Dawley大鼠根据体重随机分为对照组、模型组和菟丝子-熟地黄组。对照组每天灌胃10 mL/kg的0.9%生理盐水。模型组（GTW组）每天灌胃12 mg/kg GTW。菟丝子-熟地黄组（TSZSDH组）按模型组给药剂量每天灌胃1.56 g/kg的菟丝子-熟地黄颗粒。连续灌胃12周后测定血清黄体生成素、卵泡刺激素、雌二醇和睾酮水平，并观察睾丸组织的病理损伤。通过定量蛋白质组学评估差异表达蛋白，并通过蛋白质免疫印迹法（WB）和实时定量聚合酶链反应（RT-qPCR）进行验证。菟丝子-熟地黄可有效减轻GTW诱导的睾丸组织病理损伤。TSZSDH组和模型组共鉴定出216个差异表达蛋白。高通量蛋白质组学显示，差异表达蛋白与过氧化物酶体增殖物激活受体（PPAR）信号通路、蛋白质消化吸收及癌症中的蛋白质聚糖途径密切相关。菟丝子-熟地黄可显著上调Acsl1、Plin1、Dbil5、Plin4、Col12a1、Col1a1、Col5a3、Col1a2、Dcn的蛋白表达，从而对睾丸组织起到保护作用。通过WB和RT-qPCR实验验证了PPAR信号通路上的Acsl1、Plin1和PPARγ，发现与蛋白质组学分析结果一致。菟丝子和熟地黄可能通过调节PPAR信号通路介导的Acsl1、Plin1和PPARγ来减轻GTW对雄性大鼠睾丸组织的损伤。
