目的 基于NF-κB/NLRP3/Caspase-1信號軸探究白藜蘆醇對痛風(fēng)性腎病模型大鼠的腎臟的保護(hù)作用機(jī)制。方法 將60只SD雄性大鼠隨機(jī)分為對照組、模型組、秋水仙堿（陽性對照，0.03 mg · kg^-1）組和白藜蘆醇高、中、低劑量（1 000、500、250 mg·kg^-1）組，連續(xù)7 d ig給藥，給藥過程中，除對照組外，其余各組使用氧嗪酸鉀合并尿酸鈉的方法制備大鼠痛風(fēng)性腎病模型。ELISA法檢測大鼠血清中白細(xì)胞介素（IL）-1β、IL-18、尿酸、肌酐（SCr）、尿素氮（BUN）水平，腎臟組織勻漿中腫瘤壞死因子-α（TNF-α）、單核細(xì)胞趨化蛋白-1（MCP-1）、環(huán)氧合酶-2（COX-2）水平；HE、Masson染色觀察腎臟組織細(xì)胞形態(tài)變化；PAS染色檢測大鼠腎組織中腎小球損傷情況，TUNEL觀察腎臟組織細(xì)胞DNA損傷情況；實(shí)時熒光定量PCR （qRT-PCR）、免疫組化法檢測腎臟組織中核因子-κB （NF-κB）、NOD樣受體熱蛋白結(jié)構(gòu)域相關(guān)蛋白3（NLRP3）、半胱氨酸蛋白酶-1（Caspase-1） mRNA以及蛋白的表達(dá)量，最后采用分子對接研究白藜蘆醇與NF-κB、NLRP3、Caspase-1的結(jié)合情況。結(jié)果 與模型組比較，秋水仙堿組及白藜蘆醇各給藥組大鼠血清中IL-1β、IL-18、SCr、BUN及腎臟TNF-α、MCP-1、COX-2水平顯著降低（P<0.01），白藜蘆醇高劑量組尿酸、中和高劑量組BUN顯著降低（P<0.05）；白藜蘆醇各劑量組不同程度降低腎組織中膠原纖維化面積、腎小球陽性率以及腎組織細(xì)胞TUNEL染色陽性率，減緩病理損傷情況，其中高劑量組作用最顯著（P<0.05）；qRT-PCR、免疫組化結(jié)果表明，白藜蘆醇各給藥組均抑制腎臟組織細(xì)胞中NF-κB、NLRP3、Caspase-1mRNA和蛋白的表達(dá)，其中高劑量組作用最顯著（P<0.05）；分子對接結(jié)果進(jìn)一步表明，白藜蘆醇與NF-κB、NLRP3、Caspase-1結(jié)合狀態(tài)良好，即白藜蘆醇對NF-κB、NLRP3、Caspase-1具有良好的靶向調(diào)控作用。結(jié)論 白藜蘆醇對痛風(fēng)性腎病模型大鼠的腎臟保護(hù)作用可能為抑制NF-κB信號通路，進(jìn)而抑制NLRP3的激活從而阻斷Caspase-1招募IL-1β、IL-18，降低其分泌，遏制腎臟細(xì)胞程序性死亡的初始階段細(xì)胞焦亡的發(fā)生，從而逆轉(zhuǎn)痛風(fēng)性腎病大鼠腎組織的炎癥損傷。

Objective Exploring the protective mechanism of resveratrol on rat kidney in gouty nephropathy model based on NF-κB/NLRP 3/Caspase-1 signaling axis. Method Sixty male SD rats were randomly divided into control group, model group, colchicine (positive control, 0.03 mg· kg^-1) group, and resveratrol high, medium, and low dose (1 000, 500, 250 mg· kg^-1) group. The rats were administered ig for seven consecutive days. During the administration process, except for the control group, the other groups were prepared with potassium oxazinate combined with sodium urate to create a rat model of gouty nephropathy. Level of interleukin (IL)-1β, IL-18, uric acid, creatinine (SCr), urea nitrogen (BUN) in serum, and tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1) and cyclooxygenase-2 (COX-2) levels in renal tissue homogenate were detected by ELISA method. The morphological changes of kidney tissues were observed by HE and Masson staining. The glomerular injury in rat renal tissue was detected by PAS staining. The DNA damage in kidney tissue cells was observed by TUNEL. The expression levels of NF-κB, NLRP3, Caspase-1 mRNA and protein in kidney tissues were detected by qRT-PCR and immunohistochemistry. Finally, molecular docking technology was used to study the combination between resveratrol and NF-κB, NLRP3, Caspase-1. Result Compared with model group, the levels of IL-1β, IL-18, SCr, and BUN in the serum and and TNF-α, MCP-1 and COX-2 in renal of rats treated with colchicine and resveratro were significantly reduced (P< 0.01). While uric acid in the high-dose group of resveratrol, and BUN in medium and high dose group showed a significant decrease (P< 0.05). Each dose group of resveratrol reduced the collagen fibrosis area, glomerular positivity rate, and TUNEL staining positivity rate of renal tissue cells to varying degrees, slowing down pathological damage, and the high-dose group had the most significant effect among them (P< 0.05). qRT-PCR and immunohistochemistry results showed that all treatment groups of resveratrol inhibited NF-κB, NLRP3, and Caspase-1 mRNA and protein in renal tissue cells, with the high-dose group having the most significant effect (P< 0.05). The molecular docking results further indicate that resveratrol interacts with NF-κB. The binding state of NLRP3 and Caspase-1 is good, indicating that resveratrol has an effect on NF-κB. NLRP3 and Caspase-1 have good targeted regulatory effects. The molecular docking results further indicated that, resveratrol binds well to NF-κB, NLRP3, and Caspase-1, resveratrol showed good targeted regulation of NF-κB, NLRP3 and Caspase-1. Conclusion The renal protective effect of resveratrol in GN model of gouty arthritis with hyperuricemia is related to the inhibition of inflammatory factor secretion. Its anti-inflammatory mechanism may be through the inhibition of NF-κB signaling pathway, and then NLRP3 activation to block Caspase-1 recruitment of IL-1 IL-1 and IL-18, reduce the secretion of inflammatory factors such as IL-1β and IL-18, and curb the occurrence of cell pyroptosis in the initial stage of programmed renal cell death, thus reversing the inflammatory injury of renal tissue in GN.

R285.5

國家自然科學(xué)基金資助項(xiàng)目（82074149）；黑龍江省自然科學(xué)基金項(xiàng)目（JJ2020ZD0087）