Ochratoxin A induces nephrotoxicity in vitro and in vivo via pyroptosis
Ochratoxin A induces nephrotoxicity in vitro and in vivo via pyroptosis
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DOI：
10.1007/s00204-021-02993-6
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发表时间：
2021-02-04
影响因子：
6.1
通讯作者：
Chen, Xingxiang
中科院分区：
文献类型：
--
作者：
Li, Hu;Mao, Xinru;Chen, Xingxiang
赭曲霉毒素A（OTA）是一种广泛存在于食品和饲料中的具有肾毒性的真菌毒素污染物，可引起肾脏损伤。为了揭示连续给予OTA的肾毒性并研究与焦亡相关的潜在机制，雄性C57 BL/6小鼠腹腔注射1.0和2.0 mg/kg B.W.每隔一天OTA，共14天。在2.0 mg/kg B.W.与对照组相比，OTA给药显著增加了组织学损伤和肾纤维化分子（α-SMA、波形蛋白、TGF-β），激活了NOD样受体蛋白3（NLRP 3）炎性小体，并诱导了焦亡。在体外试验中，将Madin-Darby犬肾（MDCK）上皮细胞在无血清培养基中暴露于0-4.0 μ g/ml OTA 24小时。数据显示OTA剂量依赖性地影响细胞活力并显著上调肾纤维化基因（α-SMA、波形蛋白、TGF-β）。2.0μ g/ml OTA显著诱导NLRP 3炎性小体活化和半胱天冬酶-1依赖性焦亡，增加MDCK细胞中促炎细胞因子（IL-6、TNF-α）和焦亡相关基因（GSDMD、IL-1 β、IL-18）的表达和分泌。在用抑制剂MCC 950抑制NLRP 3激活并用小干扰RNA（siRNA）沉默NLRP 3后，这些结果显着消除。此外，半胱天冬酶-1的敲低也通过抑制焦亡来改善OTA诱导的肾纤维化。总的来说，所选剂量的OTA通过体外和体内的NLRP 3炎性体活化和半胱天冬酶-1依赖性焦亡引发肾毒性。
Ochratoxin A (OTA), a prevalent nephrotoxic mycotoxin contaminant in food and feedstuff, has been reported to induce renal injury. To disclose the nephrotoxicity of continuous administration of OTA and to investigate potential mechanisms related to pyroptosis, male C57BL/6 mice were intraperitoneally injected with 1.0 and 2.0 mg/kg B.W. OTA every other day for 14 days. At 2.0 mg/kg B.W. OTA administration significantly increased histological injury and renal fibrosis molecules (alpha-SMA, Vimentin, TGF-beta) and activated the NOD-like receptor protein 3 (NLRP3) inflammasome and induced pyroptosis compared with control. In the in vitro tests, Madin-Darby canine kidney (MDCK) epithelial cells were exposed to 0-4.0 mu g/ml OTA for 24 h in serum-free medium. Data showed that OTA dose-dependently affected cell viability and significantly up-regulated renal fibrosis genes (alpha-SMA, Vimentin, TGF-beta). 2.0 mu g/ml OTA significantly induced NLRP3 inflammasome activation and caspase-1-dependent pyroptosis, increasing the expression and secretion of pro-inflammatory cytokines (IL-6, TNF-alpha) and pyroptosis-related genes (GSDMD, IL-1 beta, IL-18) in MDCK cells. These outcomes were significantly abrogated after inhibiting NLRP3 activation with inhibitor MCC950 and silencing NLRP3 with small interfering RNA (siRNA). Furthermore, knockdown of caspase-1 also ameliorated OTA-induced renal fibrosis via the inhibition of pyroptosis. Collectively, the chosen doses of OTA-triggered nephrotoxicity through NLRP3 inflammasome activation and caspase-1-dependent pyroptosis both in vitro and in vivo.