Data for: Radiation of the Urinary Bladder Attenuates the Development of Lipopolysaccharide-Induced Cystitis

In the present study we assessed how ionization radiation affects TLR4-stimulated immune activation in lipopolysaccharide (LPS)-induced cystitis. LPS or saline was administered intravesically to female rats followed by urinary bladder irradiation (20 Gy) 24 hours later or sham treatment. Presence in the urinary bladder of inflammatory cells (mast cells, CD3+, ionized calcium-binding adapter molecule 1 (Iba-1)+, CD68+, CD40+, CD80+, CD11c+ and CD206+ cells) and expression of oxidative stress (8-OHdG), hypoxia (HIF1α) and anti-oxidative responses (NRF2, HO-1, SOD1, SOD2, catalase) were assessed 14 days later with western blot, qPCR and/or immunohistochemistry. LPS stimulation resulted in a decrease of Iba-1+ cells in the urothelium, an increase in mast cells in the submucosa and a decrease in the bladder protein expression of HO-1, while no changes in the bladder expression of 8-OHdG, NRF2, SOD1, SOD2, catalase and HIF1α were observed. Bladder irradiation inhibited the LPS-driven increase in mast cells and the decrease in Iba1+ cells. Combining LPS and radiation increased the expression of 8-OHdG and number of CD3-positive cells in the urothelium and led to a decrease in NRF2α gene expression in the urinary bladder. In conclusion, irradiation may attenuate LPS-induced immune responses in the urinary bladder but potentiates LPS-induced oxidative stress, which as a consequence may have an impact on the urinary bladder immune sensing of pathogens and danger signals.

本研究旨在探究电离辐射（ionization radiation）对脂多糖（LPS）诱导的膀胱炎中Toll样受体4（TLR4）触发的免疫激活的影响。我们向雌性大鼠膀胱内灌注给予脂多糖或生理盐水，24小时后对其膀胱实施20 Gy电离辐射或假照射处理；于造模后14天，通过蛋白质印迹（Western Blot）、实时定量聚合酶链反应（qPCR）及/或免疫组织化学法（immunohistochemistry），检测大鼠膀胱组织内炎症细胞[包括肥大细胞、CD3+细胞、离子钙结合衔接分子1阳性（Iba-1+）细胞、CD68+细胞、CD40+细胞、CD80+细胞、CD11c+细胞及CD206+细胞]的浸润情况，同时检测氧化应激标志物8-羟基脱氧鸟苷（8-OHdG）、缺氧相关因子缺氧诱导因子1α（HIF1α）以及抗氧化应答相关分子（NRF2、HO-1、SOD1、SOD2、过氧化氢酶（catalase））的表达水平。结果显示，脂多糖刺激可导致膀胱上皮内Iba-1+细胞数量减少，黏膜下层肥大细胞数量增多，并降低膀胱组织中HO-1的蛋白表达，但未观察到膀胱组织中8-OHdG、NRF2、SOD1、SOD2、过氧化氢酶及HIF1α的表达出现显著变化；膀胱照射可抑制脂多糖诱导的肥大细胞增多及Iba-1+细胞减少现象；联合脂多糖刺激与膀胱照射可上调膀胱上皮内8-OHdG的表达及CD3+细胞数量，并降低膀胱组织中NRF2的基因表达水平。综上，电离辐射可减弱膀胱组织中脂多糖诱导的免疫应答，但会增强脂多糖引发的氧化应激，这一效应可能会对膀胱识别病原体与危险信号的免疫感知功能产生影响。