Rana Mohammad Nasiruddin, Lu Jie, Xue Enfu, Ruan Jingjing, Liu Yuting, Zhang Lejun, Dhar Rana, Li Yajun, Hu Zhengqiang, Zhou Jie, Ma Wangqian, Tang Huifang
Department of Pharmacology, School of Basic Medical Sciences, Zhejiang University, Hangzhou, China.
Clinical Laboratory, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Front Pharmacol. 2021 Apr 8;12:643215. doi: 10.3389/fphar.2021.643215. eCollection 2021.
Ulcerative colitis (UC) is a form of inflammatory bowel disease, which manifests as irritation or swelling and sores in the large intestine in a relapsing and remitting manner. In a dextran sulfate sodium sulfate (DSS)-induced UC model in female mice, we found that the levels of cyclic guanosine monophosphate (cGMP) are reduced, while the expression of phosphodiesterase 9A (PDE9A) is highest among all phosphodiesterase (PDEs). Since PDE9 has the highest affinity toward cGMP, we evaluated the selective PDE9 inhibitor PF-04447943 (PF) as a potential candidate for UC treatment. PF has been extensively studies in cognitive function and in sickle cell disease, but not in models for inflammatory bowel disease (IBD). Therefore, we used female C57BL/6 mice treated with 3% DSS alone or co-treated with PF or sulfasalazine (SASP) to study the body weight, colon length, histopathology, and measure superoxide dismutase (SOD), malondialdehyde (MDA), and cGMP level, as well as cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-17 (IL-17), interleukin-12/23 (IL-12/23), interleukin-10 (IL-10), and pathways including nuclear factor kappa B (NF-κB), signal transducer and activator of transcription 3 (STAT3), and inflammasome activation. In addition, the number of dendritic cells (DC) and regulatory T cells (Treg cell) was assessed in the spleen, lymph node, and colon using flow cytometry. DSS reduced the number of goblet cells, decreased colon lengths and body weights, all of them were attenuated by PF treatment. It also suppressed the elevated level of inflammatory cytokines and increased level the anti-inflammatory cytokine, IL-10. PF treatment also reduced the DSS-induced inflammation by suppressing oxidative stress, NF-κB, STAT3, and inflammasome activation, by upregulating nuclear factor erythroid 2-related factor 2 (Nrf-2) and its downstream proteins via extracellular signal-regulated kinase (ERK) phosphorylation. Importantly, PF reversed imbalance in Treg/T helper 17 cells (Th17) cells ratio, possibly by regulating dendritic cells and Treg developmental process. In summary, this study shows the protective effect of a PDE9A inhibitor in ulcerative colitis by suppressing oxidative stress and inflammation as well as reversing the Treg/Th17 cells imbalance.
溃疡性结肠炎(UC)是炎症性肠病的一种形式,表现为大肠反复出现刺激、肿胀和溃疡。在雌性小鼠的葡聚糖硫酸钠(DSS)诱导的UC模型中,我们发现环磷酸鸟苷(cGMP)水平降低,而磷酸二酯酶9A(PDE9A)的表达在所有磷酸二酯酶(PDEs)中最高。由于PDE9对cGMP具有最高亲和力,我们评估了选择性PDE9抑制剂PF-04447943(PF)作为UC治疗的潜在候选药物。PF已在认知功能和镰状细胞病方面进行了广泛研究,但尚未在炎症性肠病(IBD)模型中进行研究。因此,我们使用单独用3% DSS处理或与PF或柳氮磺胺吡啶(SASP)联合处理的雌性C57BL/6小鼠来研究体重、结肠长度、组织病理学,并测量超氧化物歧化酶(SOD)、丙二醛(MDA)和cGMP水平,以及细胞因子如肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-17(IL-17)、白细胞介素-12/23(IL-12/23)、白细胞介素-10(IL-10),以及包括核因子κB(NF-κB)、信号转导和转录激活因子3(STAT3)和炎性小体激活在内的信号通路。此外,使用流式细胞术评估脾脏、淋巴结和结肠中树突状细胞(DC)和调节性T细胞(Treg细胞)的数量。DSS减少了杯状细胞数量,缩短了结肠长度并降低了体重,而PF处理减轻了所有这些变化。它还抑制了炎性细胞因子水平的升高,并增加了抗炎细胞因子IL-10的水平。PF处理还通过抑制氧化应激、NF-κB、STAT3和炎性小体激活,通过细胞外信号调节激酶(ERK)磷酸化上调核因子红细胞2相关因子2(Nrf-2)及其下游蛋白,从而减轻了DSS诱导的炎症。重要的是,PF可能通过调节树突状细胞和Treg发育过程逆转了Treg/辅助性T细胞17(Th17)细胞比例的失衡。总之,本研究表明PDE9A抑制剂通过抑制氧化应激和炎症以及逆转Treg/Th17细胞失衡对溃疡性结肠炎具有保护作用。