Effect of Suan Zao Ren (Semen Ziziphi Spinosae) Extract on the TXNIP/NLRP3 Pathway in Insomniac Rats  

酸枣仁提取物对失眠大鼠TXNIP/NLRP3通路的影响

作　　者：Zijing Xu Wei Xiong Qian Wang Shuyu Li Dexian Jia 许子敬;熊伟;王谦;李姝玉;贾德贤(北京中医药大学,北京)

机构地区：[1]College of Chinese Medicine,Beijing University of Chinese Medicine,Beijing,China

出　　处：《Chinese Medicine and Natural Products》2024年第3期126-135,共10页中医学报（英文）

基　　金：Research on the Mechanism of Suan Zao Ren’s(SZS)Hypnotic Effect Based on TXNIP/NLRP3 Signaling Pathway(2023-ZXFZJJ-JW-071).

摘　　要：Objectives This study aimed to investigate the therapeutic effects of Suan Zao Ren(Semen Ziziphi Spinosae,SZS)extract on insomnia induced by p-chlorophenylalanine(PCPA)in rats and its influence on the thioredoxin-interacting protein(TXNIP)/nucleotide-binding domain Leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3)inflammasome pathway,and to preliminarily explore themechanism by which SZS extract improves insomnia.Methods Fifty male Sprague–Dawley(SD)rats were used,with 8 rats in the blank group and 42 rats in the modeling group.The modeling group was induced by intraperitoneal injection of PCPA at a dose of 500mg·kg^(-1) for six consecutive days,with daily cage exchange.After 6 days,40 successfully modeled rats were randomly divided into five groups:themodel group(equal volume of distilled water),the positive group(0.75 mg·kg^(-1)),and low-,medium-,and high-dose SZS extract groups(1.5,3,and 6 g·kg^(-1),respectively),with 8 rats in each group.Treatments were administered for seven consecutive days.Enzyme-linked immunosorbent assay was used to measure levels of 5-hydroxytryptamine(5-HT)and gamma-aminobutyric acid(GABA)in the rat cerebral cortex.The thiobarbituric acid(TBA)method was used to determine malondialdehyde(MDA)levels,and the hydroxylamine method was used to determine superoxide dismutase(SOD)levels.The 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)method was used to measure total antioxidant capacity(TAOC)in the cerebral cortex.Pathological changes in the cerebral cortex were observed,and Western blot was used to detect the protein expressions of TXNIP,NLRP3,apoptosis-associated speck-like protein containing a Caspase activation and recruitment domain(CARD),and cysteine–aspartate-specific protease 1(Caspase-1)in the cerebral cortex.Results Compared with the blank group,the model group showed a significantly prolonged sleep latency(p<0.001)and a significantly shortened sleep duration(p<0.001).There were no changes in serum MDA and SOD levels.MDA levels in the cerebral cortex目的:探讨酸枣仁提取物对对氯苯丙氨酸(p-chlorophenylalanine,PCPA)诱导失眠大鼠的治疗效果及其对硫氧还蛋白相互作用蛋白(thioredoxin-interacting protein,TXNIP)/核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3(nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3,NLRP3)炎症通路的影响,并初步探讨酸枣仁提取物改善失眠的机制。方法:50只SD大鼠随机分为空白组8只,建模组42只。建模组连续6天通过腹腔注射500 mg·kg^(-1)对氯苯丙氨酸建立失眠模型,并每日更换笼舍。6天后,40只成功建模的大鼠随机分为五组:模型组(等体积蒸馏水)、佐匹克隆组(0.75 mg·kg^(-1))及高(6 g·kg^(-1))、中(3 g·kg^(-1))、低(1.5 g·kg^(-1))剂量酸枣仁提取物组,每组8只大鼠。治疗持续7天。采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定大鼠大脑皮层中5-羟色胺(5-hydroxytryptamine,5-HT)和γ氨基丁酸(γaminobutyric acid,GABA)的水平;采用硫代巴比妥酸法(thiobarbituric acid method,TBA)测定丙二醛(malondialdehyde,MDA)水平;采用羟胺法测定超氧化物歧化酶(superoxide dismutase,SOD)水平;使用2,2′-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐[2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS]测定脑皮层总抗氧化能力(total antioxidant capacity,TAOC);观察脑皮层的病理变化,并采用蛋白免疫印迹法(western blot,WB)检测脑皮层中TXNIP、NLRP3、凋亡相关斑点样蛋白质(apoptosis-associated speck-like protein containing a CARD,ASC)及半胱氨酸天冬氨酸蛋白酶1(cysteine-aspartate-specific protease,Caspase-1)的蛋白表达水平。结果:与空白组相比,模型组睡眠潜伏期显著延长(P<0.001),睡眠持续时间显著缩短(P<0.001),血清中MDA和SOD水平无明显变化;脑皮层中MDA水平显著增加(P<0.001),而SOD和TAOC水平显著降低(P<0.001);5-HT水平显著增加(P<0.05),GABA水平显著降低(P<0.001)。与

关 键 词：INSOMNIA Suan Zao Ren TXNIP/NLRP3 oxidative stress sedative and hypnotic 

分 类 号：R285.5[医药卫生—中药学]