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麻黄草通过上调促性腺激素释放激素信号通路减轻阿霉素诱导的睾丸毒性。

Ephedra herb reduces adriamycin-induced testicular toxicity by upregulating the gonadotropin-releasing hormone signalling pathway.

机构信息

Department of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China; The Engineering and Technology Center for Chinese Medicine Development of Henan Province, Zhengzhou 450046, China.

Department of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China.

出版信息

Biomed Pharmacother. 2022 Jun;150:113061. doi: 10.1016/j.biopha.2022.113061. Epub 2022 May 23.

Abstract

OBJECTIVE

We investigated the protective effects of ephedra herb (HEPH) on adriamycin-induced testicular toxicity in rats and explored the potential mechanisms underlying these effects.

METHODS

A rat model of adriamycin injury was established, and sperm motility-related indicator and oxidative stress levels in the testis were evaluated. Serum levels of sex hormones and levels of testicular cell apoptosis were detected by enzyme-linked immunosorbent assay and flow cytometry, respectively. Western blotting (WB), immunofluorescence analyses, and reverse transcription-polymerase chain reaction (RT-PCR) were performed to evaluate the gonadotropin-releasing hormone (GnRH) signalling pathway- and meiosis-related genes and proteins. In subsequent in vitro experiments, adriamycin was used to stimulate GC-1 cells, which were treated with HEPH, ephedrine, or pseudoephedrine. Cell viability was assessed using flow cytometry to detect apoptosis and reactive oxygen species, whereas the GnRH signalling pathway and levels of meiosis-related genes and proteins were evaluated by InCell WB, a high-content imaging system, and RT-PCR.

RESULTS

Per in vivo experiments, HEPH restored testicular weight and function, sperm characteristics, serum and tissue hormonal levels, and antioxidant defences and significantly activated the GnRH signalling pathway- and meiosis-related protein levels. All protective effects of HEPH against adriamycin-induced injury were antagonised by the GnRH antagonist cetrorelix. In vitro, HEPH, ephedrine, and pseudoephedrine significantly reduced adriamycin-induced GC-1 cell apoptosis and reactive oxygen species levels and increased the expression of GnRH signalling pathway- and meiosis-related proteins. The effect of pseudoephedrine was greater than that of ephedrine, and these findings may be an important basis for understanding the effects of HEPH.

摘要

目的

研究麻黄草(HEPH)对阿霉素诱导的大鼠睾丸毒性的保护作用,并探讨其潜在作用机制。

方法

建立阿霉素损伤大鼠模型,评估精子运动相关指标和睾丸氧化应激水平。采用酶联免疫吸附试验和流式细胞术检测血清性激素水平和睾丸细胞凋亡水平,采用Western blot(WB)、免疫荧光分析和逆转录-聚合酶链反应(RT-PCR)评估促性腺激素释放激素(GnRH)信号通路和减数分裂相关基因和蛋白。随后,在体外实验中,用阿霉素刺激 GC-1 细胞,并用 HEPH、麻黄碱或伪麻黄碱处理。采用流式细胞术评估细胞活力以检测细胞凋亡和活性氧,采用 InCell WB、高内涵成像系统和 RT-PCR 评估 GnRH 信号通路和减数分裂相关基因和蛋白水平。

结果

体内实验中,HEPH 恢复了睾丸重量和功能、精子特征、血清和组织激素水平以及抗氧化防御功能,显著激活了 GnRH 信号通路和减数分裂相关蛋白水平。HEPH 对阿霉素诱导损伤的所有保护作用均被 GnRH 拮抗剂 cetrorelix 拮抗。在体外,HEPH、麻黄碱和伪麻黄碱显著降低了阿霉素诱导的 GC-1 细胞凋亡和活性氧水平,增加了 GnRH 信号通路和减数分裂相关蛋白的表达。伪麻黄碱的作用大于麻黄碱,这些发现可能是理解 HEPH 作用的重要基础。

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