Khanmohammad Khadije Rezai, Khalili Mohammad Bagher, Sadeh Maryam, Talebi Ali Reza, Astani Akram, Shams Ali, Zare Fateme
Department of Microbiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Department of Laboratory Sciences, Faculty of Paramedical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Clin Exp Reprod Med. 2021 Jun;48(2):105-110. doi: 10.5653/cerm.2020.03888. Epub 2021 May 21.
Uropathogenic Escherichia coli is known to cause urinary tract infections, and the endotoxin (lipopolysaccharide [LPS]) of this bacterium may cause deficiencies of sperm quality and morphology. In the present study, the effects of LPS on mouse sperm were studied, and the levels of interleukin (IL)-17A and possible changes in testis tissue were evaluated.
LPS of uropathogenic E. coli was extracted using the methanol-chloroform method, followed confirmation using sodium dodecyl sulfate-polyacrylamide electrophoresis. Purified LPS (100 µg/kg) or phosphate-buffered saline was injected intraperitoneally into BALB/c mice for 7 days consecutively in the test and control groups, Mice were sacrificed on days 3, 7, and 42 after the first injection. Blood was tested for levels of IL-17A using the enzyme-linked immunosorbent assay method. Testis tissue and sperm were collected from each mouse and were studied according to standard protocols.
The mean sperm count and motility significantly decreased (p=0.03) at 3, 7, and 42 days after the injections. The level of IL-17A in the test groups increased, but not significantly (p=0.8, p=0.11, and p=0.15, respectively). Microscopic studies showed no obvious changes in the morphology of the testis tissue; however, significant changes were observed in the cellular parenchyma on day 42.
LPS can stimulate the immune system to produce proinflammatory cytokines, resulting in an immune response in the testis and ultimately leading to deficiency in sperm parameters and testis tissue damage. In addition, the presence of LPS could significantly impair sperm parameters, as shown by the finding of decreased motility.
已知尿路致病性大肠杆菌会引起尿路感染,该细菌的内毒素(脂多糖[LPS])可能导致精子质量和形态缺陷。在本研究中,研究了LPS对小鼠精子的影响,并评估了白细胞介素(IL)-17A水平以及睾丸组织可能发生的变化。
采用甲醇-氯仿法提取尿路致病性大肠杆菌的LPS,随后用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行确认。在试验组和对照组中,将纯化的LPS(100μg/kg)或磷酸盐缓冲盐水连续7天腹腔注射到BALB/c小鼠体内。在首次注射后的第3、7和42天处死小鼠。采用酶联免疫吸附测定法检测血液中IL-17A水平。从每只小鼠收集睾丸组织和精子,并按照标准方案进行研究。
注射后第3、7和42天,平均精子计数和活力显著下降(p=0.03)。试验组中IL-17A水平升高,但差异不显著(分别为p=0.8、p=0.11和p=0.15)。显微镜研究显示睾丸组织形态无明显变化;然而,在第42天观察到细胞实质有显著变化。
LPS可刺激免疫系统产生促炎细胞因子,导致睾丸发生免疫反应,最终导致精子参数缺陷和睾丸组织损伤。此外,LPS的存在可显著损害精子参数,如活力下降所示。