c-Jun氨基末端蛋白激酶在人角膜上皮细胞调控NLRP3炎症小体激活中的作用

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Abstract Objective: To investigate the role of JNK (c-Jun N-terminal kinase) in hyperosmosis-induced NLRP3 (NLR family pyrin domain containing 3) up-regulation and IL-1β secretion in the human corneal epithelial cells. Methods: HCECs cultured in normal osmolar media (310 mOsm) were switched to 500 mOsm, with or without the JNK inhibitor SP600125 (20 μmol/L). The mRNA levels of JNK, NLRP3, ASC, caspase-1 and IL-1β were detected by RT-qPCR. Caspase-1 activity was tested by caspase-1 colorimetric assay kits. Cell apoptosis was tested by AOEB staining method. The protein expression of NLRP3 and IL-1β were detected by immunofluorescence staining. And the protein level of JNK, p-JNK, NLRP3, caspase-1, ASC and IL-1β were determined by Western blot. Results: Hyperosmosis-induced mRNA overexpressions of JNK1, JNK2, NLRP3, caspase-1, IL-1β, TNF-α, IL-8 and IL-6 were significantly inhibited by JNK inhibitor (P<0.05). Moreover, hyperosmosis-induced caspase-1 activity was decreased with JNK inhibitor treatment by caspase-1 colorimetric assay (P<0.001). JNK inhibitor was able to downregulate cell apoptosis by AOEB staining. The protein overexpressions of NLRP3 and IL-1β induced by hyperosmosis were reversed by JNK inhibitor with immunofluorescence staining assays. And the protein overexpressions of p-JNK, NLRP3, caspase-1 and IL-1β induced by hyperosmosis were also downregulated with the use of JNK inhibitor by Western blot (P<0.05). Conclusion: NLRP3 inflammasome activation and subsequent IL-1β secretion are key consecutive events in HCECs response to hyperosmotic stress, and JNK may play a pivotal role in the regulation of the inflammasomes activation and expression of inflammatory factors.

Key words： hyperosmotic c-Jun N-terminal kinase inflammasome interleukin-1beta xerophthalmia

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	TAN Qiufan
	CHEN Huaicheng
	ZHU Yirui
	ZHU Hanlei
	ZHENG Qinxiang
	CHEN Wei

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2018/V48/I1/13

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