目的通过观察补阳还五汤对高血压病大鼠硫氧还蛋白互作蛋白-消皮素D(TXNIP/GSDMD)信号转导通路的影响,从焦亡角度探讨补阳还五汤对高血压病大鼠主动脉血管内皮损伤的保护机制。方法将大鼠随机分为空白对照组,模型组,补阳还五汤低、中、...目的通过观察补阳还五汤对高血压病大鼠硫氧还蛋白互作蛋白-消皮素D(TXNIP/GSDMD)信号转导通路的影响,从焦亡角度探讨补阳还五汤对高血压病大鼠主动脉血管内皮损伤的保护机制。方法将大鼠随机分为空白对照组,模型组,补阳还五汤低、中、高剂量组,氢氯噻嗪组,每组10只,所有大鼠于SPF级动物实验中心普通饲料适应性饲养7 d。空白对照组每日给予低盐饲料喂养,其余各组采用高盐饲料喂养,同时,补阳还五汤低、中、高剂量组分别以0.275、0.55、1.1 g/mL浓度中药干预,氢氯噻嗪组以10 mg/(kg·d)浓度药物干预。3周后,观察到大鼠血压比造模前升高大于或等于30 mm Hg(1 mm Hg≈0.133 kPa),且收缩压(SBP)≥160 mm Hg即高血压病大鼠造模成功。取大鼠主动脉进行检测,HE染色观察大鼠主动脉病理形态改变;采用ELISA检测大鼠主动脉中血管紧张素Ⅱ(AngⅡ)、白细胞介素18(IL-18)、白细胞介素1β(IL-1β)、超氧化物歧化酶(superoxide dismutase,SOD)以及丙二醛(malondialdehyde,MDA)蛋白表达;荧光定量PCR检测大鼠主动脉中TXNIP、GSDMD、还原性辅酶Ⅱ氧化酶4(NOX4)、IL-1β的mRNA表达,免疫印迹实验(Western blot)检测大鼠主动脉中GSDMD的蛋白表达,免疫组化(IHC)检测大鼠主动脉中TXNIP、NOD样受体蛋白3(NLRP3)、GSDMD的蛋白表达。结果与空白对照组相比,模型组大鼠血压明显升高,主动脉内外膜都有一定脱落,界线模糊,血管中膜弹性纤维结构比较疏松,排列有一定紊乱;TXNIP、GSDMD、IL-1β、NOX4的mRNA表达上升(P<0.05);TXNIP、NLRP3、GSDMD、IL-1β、IL-18、AngⅡ、MDA的蛋白表达升高(P<0.05),SOD蛋白表达降低(P<0.05)。与模型组相比,补阳还五汤中剂量组和氢氯噻嗪组大鼠血压明显降低,血管内外膜更加完整、光滑,结构更加致密,界线较为清晰明显,补阳还五汤低、高剂量组内外膜仍有一定脱落,弹性纤维排列比较杂乱,变化不明显;药物干预后各组TXNIP、GSDMD、IL-1β、NOX4的mRNA表达均下降(P<0.05);补阳还五汤中剂量组和氢氯噻嗪组TXNIP、NLRP3、GSDMD、IL-1β、IL-18、AngⅡ、MDA的蛋白表达降低(P<0.05),SOD蛋白表达升高(P<0.05)。结论补阳还五汤可通过抑制氧化应激,调节TXNIP/GSDMD信号通路的分子表达,调控血管内皮焦亡,从而发挥保护血管、治疗高血压病的作用。展开更多
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-inte...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 were significantly increased(p<0.001),while SOD and TAOC levels were significantly decreased(p<0.001).The 5-HT level was increased(p<0.05),and the GABA level was significantly decreased(p<0.001).SZS extract improved these conditions to varying degrees.Light microscopy showed no significant changes in cortical neurons but transmission electron microscopy revealed intact mitochondrial structures in the blank group,while the model group showed swollen and unclear mitochondria with reduced organelles.After 7 days of treatment,these conditions improved in the SZS extract groups.Compared with the blank group,the expressions of the four proteins in the model group were increased,and the expressions of these proteins were decreased in the SZS extract groups compared with the model group.Conclusion SZS extract may exert an antioxidant effect to treat insomnia by downregulating the expression of TXNIP/NLRP3 proteins and regulating oxidative stress levels in the cerebral cortex.展开更多
目的:探讨橘皮素调节硫氧还蛋白互作蛋白(thioredoxin-interacting protein,TXNIP)/含NLR家族PYRIN域蛋白3(NLR family PYRIN domain containing protein 3,NLRP3)信号通路对重症肺炎大鼠细胞焦亡的影响。方法:将SD大鼠分为对照组、重...目的:探讨橘皮素调节硫氧还蛋白互作蛋白(thioredoxin-interacting protein,TXNIP)/含NLR家族PYRIN域蛋白3(NLR family PYRIN domain containing protein 3,NLRP3)信号通路对重症肺炎大鼠细胞焦亡的影响。方法:将SD大鼠分为对照组、重症肺炎组、橘皮素低剂量组、橘皮素高剂量组、橘皮素高剂量+pcDNA组、橘皮素高剂量+pcDNA-TXNIP(TXNIP激活剂)组,每组12只。除对照组外,其它组大鼠均采用气管穿刺肺炎克雷伯菌液的方式构建重症肺炎大鼠模型。造模成功后立即给药处理,给药共持续4周。动物肺功能仪监测大鼠呼气峰流速(peak expiratory flow,PEF)、用力肺活量(forced vital capacity,FVC)的变化;ELISA检测大鼠肺泡灌洗液中白细胞介素(interleukin,IL)-1β和IL-18水平;检测大鼠肺湿干重比值变化;HE染色检测大鼠肺组织病理变化;碘化丙啶(propidium iodide,PI)染色检测大鼠肺组织中细胞焦亡;Western blot检测大鼠肺组织中TXNIP、NLRP3、裂解的天冬氨酸特异性半胱氨酸蛋白酶-1(Cleaved Cysteinyl aspartate-specific proteinase-1,Cleaved Caspase-1)、Gasdermin家族蛋白D-N端(N-terminal fragment of GSDMD,GSDMD-N)蛋白表达。结果:与对照组比较,重症肺炎组大鼠肺组织严重损伤,PEF、FVC降低,肺泡灌洗液中IL-1β和IL-18水平、肺湿干重比值升高,肺组织中细胞焦亡率及TXNIP、NLRP3、Cleaved Caspase-1、GSDMD-N蛋白表达升高(P<0.05);与重症肺炎组比较,橘皮素低剂量组、橘皮素高剂量组大鼠肺组织损伤有所改善,PEF、FVC升高,肺泡灌洗液中IL-1β和IL-18水平、肺湿干重比值降低,肺组织中细胞焦亡率及TXNIP、NLRP3、Cleaved Caspase-1、GSDMD-N蛋白表达降低(P<0.05);pcDNA-TXNIP逆转了高剂量橘皮素对重症肺炎的保护作用。结论:橘皮素可能通过阻断TXNIP/NLRP3通路抑制重症肺炎大鼠炎症及细胞焦亡。展开更多
文摘目的通过观察补阳还五汤对高血压病大鼠硫氧还蛋白互作蛋白-消皮素D(TXNIP/GSDMD)信号转导通路的影响,从焦亡角度探讨补阳还五汤对高血压病大鼠主动脉血管内皮损伤的保护机制。方法将大鼠随机分为空白对照组,模型组,补阳还五汤低、中、高剂量组,氢氯噻嗪组,每组10只,所有大鼠于SPF级动物实验中心普通饲料适应性饲养7 d。空白对照组每日给予低盐饲料喂养,其余各组采用高盐饲料喂养,同时,补阳还五汤低、中、高剂量组分别以0.275、0.55、1.1 g/mL浓度中药干预,氢氯噻嗪组以10 mg/(kg·d)浓度药物干预。3周后,观察到大鼠血压比造模前升高大于或等于30 mm Hg(1 mm Hg≈0.133 kPa),且收缩压(SBP)≥160 mm Hg即高血压病大鼠造模成功。取大鼠主动脉进行检测,HE染色观察大鼠主动脉病理形态改变;采用ELISA检测大鼠主动脉中血管紧张素Ⅱ(AngⅡ)、白细胞介素18(IL-18)、白细胞介素1β(IL-1β)、超氧化物歧化酶(superoxide dismutase,SOD)以及丙二醛(malondialdehyde,MDA)蛋白表达;荧光定量PCR检测大鼠主动脉中TXNIP、GSDMD、还原性辅酶Ⅱ氧化酶4(NOX4)、IL-1β的mRNA表达,免疫印迹实验(Western blot)检测大鼠主动脉中GSDMD的蛋白表达,免疫组化(IHC)检测大鼠主动脉中TXNIP、NOD样受体蛋白3(NLRP3)、GSDMD的蛋白表达。结果与空白对照组相比,模型组大鼠血压明显升高,主动脉内外膜都有一定脱落,界线模糊,血管中膜弹性纤维结构比较疏松,排列有一定紊乱;TXNIP、GSDMD、IL-1β、NOX4的mRNA表达上升(P<0.05);TXNIP、NLRP3、GSDMD、IL-1β、IL-18、AngⅡ、MDA的蛋白表达升高(P<0.05),SOD蛋白表达降低(P<0.05)。与模型组相比,补阳还五汤中剂量组和氢氯噻嗪组大鼠血压明显降低,血管内外膜更加完整、光滑,结构更加致密,界线较为清晰明显,补阳还五汤低、高剂量组内外膜仍有一定脱落,弹性纤维排列比较杂乱,变化不明显;药物干预后各组TXNIP、GSDMD、IL-1β、NOX4的mRNA表达均下降(P<0.05);补阳还五汤中剂量组和氢氯噻嗪组TXNIP、NLRP3、GSDMD、IL-1β、IL-18、AngⅡ、MDA的蛋白表达降低(P<0.05),SOD蛋白表达升高(P<0.05)。结论补阳还五汤可通过抑制氧化应激,调节TXNIP/GSDMD信号通路的分子表达,调控血管内皮焦亡,从而发挥保护血管、治疗高血压病的作用。
基金Research on the Mechanism of Suan Zao Ren’s(SZS)Hypnotic Effect Based on TXNIP/NLRP3 Signaling Pathway(2023-ZXFZJJ-JW-071).
文摘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 were significantly increased(p<0.001),while SOD and TAOC levels were significantly decreased(p<0.001).The 5-HT level was increased(p<0.05),and the GABA level was significantly decreased(p<0.001).SZS extract improved these conditions to varying degrees.Light microscopy showed no significant changes in cortical neurons but transmission electron microscopy revealed intact mitochondrial structures in the blank group,while the model group showed swollen and unclear mitochondria with reduced organelles.After 7 days of treatment,these conditions improved in the SZS extract groups.Compared with the blank group,the expressions of the four proteins in the model group were increased,and the expressions of these proteins were decreased in the SZS extract groups compared with the model group.Conclusion SZS extract may exert an antioxidant effect to treat insomnia by downregulating the expression of TXNIP/NLRP3 proteins and regulating oxidative stress levels in the cerebral cortex.
