LncRNA ZNF883-Mediated NLRP3 Inflammasome Activation and Epilepsy Development Involve USP47 Upregulation
LncRNA ZNF883-Mediated NLRP3 Inflammasome Activation and Epilepsy Development Involve USP47 Upregulation
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DOI：
10.1007/s12035-022-02902-7
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发表时间：
2022-06-09
影响因子：
5.1
通讯作者：
Zhang, Chen
中科院分区：
文献类型：
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作者：
Gong, Lina;Han, Yaru;Zhang, Chen
本研究的目的是探讨长链非编码RNA（lncRNA）ZNF 883调控癫痫（EP）NOD样受体3（NLRP 3）炎性小体激活的机制。分别采用毛果芸香碱和无镁细胞外液建立大鼠和细胞EP模型，检测ZNF 883、microRNA（miR）-138-5p、泛素特异性肽酶47（USP 47）和NLRP 3的差异表达。采用全细胞膜片钳技术观察海马神经元的病理变化。在癫痫神经元中ZNF 883、miR-138- 5 p和USP 47的表达被改变，并且EP大鼠被注射sh-ZNF 883。然后，测量ZNF 883、miR-138- 5 p和USP 47水平的变化。检测海马的组织病理学，沿着IL-6、IL-1 β、TNF-α和NLRP 3的检测。测定大鼠和细胞EP模型中的神经元凋亡。确定了ZNF 883、miR-138- 5 p和USP 47之间的关系以及USP 47对NLRP 3遍在化的调节。在癫痫神经元和大鼠中，ZNF 883、USP 47和NLRP 3表达增加，miR-138- 5 p表达下调，同时炎症和凋亡加重。ZNF 883过表达的癫痫神经元中USP 47的表达增加。ZNF 883靶向miR-138- 5 p，miR-138- 5 p负调控USP 47。在癫痫神经元中，抑制miR-138- 5 p或过表达USP 47部分逆转了ZNF 883沉默诱导的对NLRP 3炎性小体激活、神经元凋亡和癫痫样活动的抑制。EP大鼠中ZNF 883沉默降低USP 47和NLRP 3，增加miR-138- 5 p，并抑制炎症和凋亡。USP 47逆转了NLRP 3的泛素化。ZNF 883通过海绵状作用于miR-138- 5 p上调USP 47，抑制NLRP 3泛素化，促进EP。
The goal of this study was to characterize the mechanisms of long noncoding RNA (lncRNA) ZNF883 regulating NOD-like receptor 3 (NLRP3) inflammasome activation in epilepsy (EP). Rat and cellular EP models were established using pilocarpine and magnesium-free extracellular fluid, respectively, to detect the differential expression of ZNF883, microRNA (miR)-138-5p, ubiquitin-specific peptidase 47 (USP47), and NLRP3. The pathology of the hippocampal neurons was examined by whole-cell patch clamping. The expression of ZNF883, miR-138-5p, and USP47 was modified in epileptic neurons, and the EP rats were injected with sh-ZNF883. Then, alterations in ZNF883, miR-138-5p, and USP47 levels were measured. The histopathology of the hippocampus was detected, along with the detection of IL-6, IL-1 beta, TNF-alpha, and NLRP3. Neuronal apoptosis in the rat and cellular EP models was determined. The relationship among ZNF883, miR-138-5p, and USP47 as well as the regulation of NLRP3 ubiquitination by USP47 was determined. ZNF883, USP47, and NLRP3 were increasingly expressed and miR-138-5p was downregulated in epileptic neurons and rats, concurrent with aggravated inflammation and apoptosis. ZNF883 overexpression in epileptic neurons elevated USP47 expression. ZNF883 targeted miR-138-5p and miR-138-5p negatively regulated USP47. In epileptic neurons, inhibiting miR-138-5p or overexpressing USP47 partially reversed the ZNF883 silencing-induced inhibition on NLRP3 inflammasome activation, neuronal apoptosis, and epileptiform activity. ZNF883 silencing in EP rats decreased USP47 and NLRP3, increased miR-138-5p, and inhibited inflammation and apoptosis. USP47 reversed the ubiquitination of NLRP3. ZNF883 inhibits NLRP3 ubiquitination and promotes EP through upregulating USP47 by sponging miR-138-5p.