Department of Urology, Sapporo Medical University School of Medicine, Sapporo, 060-8556, Japan.
Department of Neural Regenerative Medicine, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, Sapporo, Hokkaido, 060-8556, Japan.
BMC Urol. 2021 Nov 13;21(1):156. doi: 10.1186/s12894-021-00923-3.
Interstitial cystitis/bladder pain syndrome (IC/BPS) categorized with and without Hunner lesions is a condition that displays chronic pelvic pain related to the bladder with no efficacious treatment options. There are strong associations suggested between Hunner-type IC and autoimmune diseases. Recently, we established an animal model of Hunner-type IC using a Toll-like receptor-7 (TLR7) agonist. Intravenous infusion of mesenchymal stem cells (MSCs) can be used to treat injury via multimodal and orchestrated therapeutic mechanisms including anti-inflammatory effects. Here, we investigated whether infused MSCs elicit therapeutic efficacy associated with the TLR7-related anti-inflammatory pathway in our Hunner-type IC model.
Voiding behaviors were monitored 24 h prior to the Loxoribine (LX), which is a TLR7 agonist instillation in order to establish a Hunner-type IC model (from - 24 to 0 h) in female Sprague-Dawley rats. LX was instilled transurethrally into the bladder. At 0 h, the initial freezing behavior test confirmed that no freezing behavior was observed in any of the animals. The LX-instilled animals were randomized. Randomized LX-instilled rats were intravenously infused with MSCs or with vehicle through the right external jugular vein. Sampling tissue for green fluorescent protein (GFP)-positive MSCs were carried out at 48 h. Second voiding behavior tests were monitored from 72 to 96 h. After the final evaluation of the freezing behavior test at 96 h after LX instillation (72 h after MSC or vehicle infusion), histological evaluation with H&E staining and quantitative real-time polymerase chain reaction (RT-PCR) to analyze the mRNA expression levels of inflammatory cytokines were performed.
Freezing behavior was reduced in the MSC group, and voiding behavior in the MSC group did not deteriorate. Hematoxylin-eosin staining showed that mucosal edema, leukocyte infiltration, and hemorrhage were suppressed in the MSC group. The relative expression of interferon-β mRNA in the bladder of the MSC group was inhibited. Numerous GFP-positive MSCs were distributed mainly in the submucosal and mucosal layers of the inflammatory bladder wall.
Intravenous infusion of MSCs may have therapeutic efficacy in a LX-instilled Hunner-type IC rat model via a TLR7-related anti-inflammatory pathway.
伴有或不伴有 Hunner 病变的间质性膀胱炎/膀胱疼痛综合征 (IC/BPS) 是一种表现为与膀胱相关的慢性盆腔疼痛的疾病,目前尚无有效的治疗方法。Hunner 型 IC 与自身免疫性疾病之间存在很强的关联。最近,我们使用 Toll 样受体 7 (TLR7) 激动剂建立了 Hunner 型 IC 的动物模型。间充质干细胞 (MSCs) 的静脉输注可通过多种方式和协调的治疗机制发挥治疗作用,包括抗炎作用。在这里,我们研究了在我们的 Hunner 型 IC 模型中,输注的 MSCs 是否通过与 TLR7 相关的抗炎途径产生治疗效果。
在使用 TLR7 激动剂洛索利宾 (LX) 进行膀胱灌注以建立 Hunner 型 IC 模型之前 24 小时(从-24 小时到 0 小时),监测排尿行为。将 LX 通过经尿道灌注到膀胱中。在 0 小时,最初的冷冻行为测试证实,没有动物表现出冷冻行为。将 LX 灌注的动物随机分组。将随机分组的 LX 灌注大鼠通过右颈外静脉静脉内输注 MSCs 或载体。在 48 小时时采集用于 GFP 阳性 MSCs 的组织样本。从 72 小时到 96 小时进行第二次排尿行为测试。在 LX 灌注后 96 小时(MSC 或载体输注后 72 小时)进行最后一次冷冻行为测试评估后,进行 H&E 染色的组织学评估和定量实时聚合酶链反应 (RT-PCR) 以分析炎症细胞因子的 mRNA 表达水平。
MSC 组的冷冻行为减少,而 MSC 组的排尿行为没有恶化。苏木精-伊红染色显示 MSC 组的黏膜水肿、白细胞浸润和出血得到抑制。MSC 组膀胱中干扰素-β mRNA 的相对表达受到抑制。大量 GFP 阳性 MSCs 主要分布在炎症性膀胱壁的黏膜下层和黏膜层。
MSC 的静脉输注可能通过 TLR7 相关的抗炎途径在 LX 灌注的 Hunner 型 IC 大鼠模型中具有治疗效果。