Huang Chao, Zhang Wenbo, Sun Aijun, Zhang Xi, Guo Jinping, Ji Ruijuan, Qiao Liang, Sun Xuejun, Zhao Dongbao
Department of Anatomy, Institute of Biomedical Engineering, Naval Medical University, #800 Xiangyin Road, Shanghai 200433, China.
Department of Rheumatology and Immunology, Changhai Hospital Affiliated to the Naval Medical University, Shanghai 200003, China.
Oxid Med Cell Longev. 2020 Aug 18;2020:7075836. doi: 10.1155/2020/7075836. eCollection 2020.
The present study is aimed at investigating the anti-inflammatory, antioxidative, and antiapoptotic effects of methane on lipopolysaccharide- (LPS-) induced acute orchitis and its potential mechanisms.
Adult male rats were intraperitoneally (i.p.) injected with methane-rich saline (MS, 20 mL/kg) following LPS (5 mg/kg, i.p.). The survival rate was assessed every 12 h until 72 h after LPS induction, and surviving rats were sacrificed for further detection. The wet/dry (/) ratio was determined, and testicular damage was histologically assessed. Inflammatory cytokines in the testes and serum, including interleukin-1 (IL-1), IL-6, IL-10, and tumor necrosis factor- (TNF-), were measured using ELISA and RT-qPCR. Oxidative stress was evaluated by the level of superoxide dismutase (SOD) and malondialdehyde (MDA). Testicular apoptosis was detected via TUNEL staining. The expression of prokineticin 2 (PK2)/prokineticin receptor 1 (PKR1) was also analyzed using RT-qPCR, western blotting, and immunohistochemistry.
It was found that methane significantly prolonged rat survival, decreased the W/D ratio, alleviated LPS-induced histological changes, and reduced apoptotic cells in the testes. Additionally, methane suppressed and promoted the production of pro- and anti-inflammatory cytokines, respectively. Furthermore, methane significantly increased SOD levels, decreased MDA levels, and decreased testicular expression of PK2 and PKR1. Therefore, methane exerts therapeutic effects on acute orchitis and might be a new and convenient strategy for the treatment of inflammation-related testicular diseases.
本研究旨在探讨甲烷对脂多糖（LPS）诱导的急性睾丸炎的抗炎、抗氧化和抗凋亡作用及其潜在机制。
成年雄性大鼠腹腔注射LPS（5mg/kg，腹腔注射）后，腹腔注射富甲烷盐水（MS，20mL/kg）。在LPS诱导后每12小时评估一次生存率，直至72小时，处死存活的大鼠进行进一步检测。测定湿/干（W/D）比值，并对睾丸损伤进行组织学评估。采用ELISA和RT-qPCR检测睾丸和血清中的炎性细胞因子，包括白细胞介素-1（IL-1）、IL-6、IL-10和肿瘤坏死因子-（TNF-）。通过超氧化物歧化酶（SOD）和丙二醛（MDA）水平评估氧化应激。通过TUNEL染色检测睾丸细胞凋亡。还采用RT-qPCR、western blotting和免疫组织化学分析前动力蛋白2（PK2）/前动力蛋白受体1（PKR1）的表达。
发现甲烷显著延长大鼠生存期，降低W/D比值，减轻LPS诱导的组织学变化，并减少睾丸中的凋亡细胞。此外，甲烷分别抑制和促进促炎和抗炎细胞因子的产生。此外，甲烷显著提高SOD水平，降低MDA水平，并降低睾丸中PK2和PKR1的表达。因此，甲烷对急性睾丸炎具有治疗作用，可能是治疗炎症相关睾丸疾病的一种新的便捷策略。
