p-nitrotoluene is an important organic intermediate widely used in pesticide, foamed plastics, dyestuff and medicine industries. In this paper, Crusian carps (Carassius auratus) were exposed to dilute p-nitrotoluene s...p-nitrotoluene is an important organic intermediate widely used in pesticide, foamed plastics, dyestuff and medicine industries. In this paper, Crusian carps (Carassius auratus) were exposed to dilute p-nitrotoluene solutions with different concentrations of 0, 20, 40, 80, 120, 160, 240 and 320 μg/L; the subsequent physiological responses to the chemical were observed, and the variation characteristics of blood corpuscles, ovaries and oocytes were studied by bio-microscopy. Results showed that even very low concentrations of p-nitrotoluene would lead to toxicosis and even the death of Carassius auratus. The bio-microscopic structures revealed that, compared with the compartment of Carassius auratus growing in the p-nitrotoluene–free water (the control group), the lower concentrations of p-nitrotoluene (for example no more than 80 μg/L) would result in the faster disintegration of erythrocytes and the significant enlargement of oocytes, while the higher contents of p-nitrotoluene (no less than 240 μg/L in this case) would inhibit the division of cells and their further growth, even degenerate blood corpuscles and ovaries, suggesting stronger estrogenic activities.展开更多
目的探讨醒脑静注射液对缺糖缺氧再灌注(oxygen and glucose deprivation/reoxygenation,OGD/R)损伤诱导SH-SY5Y细胞焦亡的保护作用及机制。方法使用不同浓度醒脑静注射液预处理SH-SY5Y细胞,确定醒脑静注射液给药浓度;将SH-SY5Y细胞随...目的探讨醒脑静注射液对缺糖缺氧再灌注(oxygen and glucose deprivation/reoxygenation,OGD/R)损伤诱导SH-SY5Y细胞焦亡的保护作用及机制。方法使用不同浓度醒脑静注射液预处理SH-SY5Y细胞,确定醒脑静注射液给药浓度;将SH-SY5Y细胞随机分为对照组、OGD/R组、醒脑静注射液+OGD/R组(10μL/mL醒脑静注射液)以及醒脑静注射液+OGD/R+核因子E2相关因子2(nuclear factor-E2 related factor 2,Nrf2)抑制剂组(10μL/mL醒脑静注射液+2μmol/mL ML385)共4组,每组实验进行6次(n=6),采用MTT比色法检测细胞存活率;采用酶联免疫吸附试验(enzyme-linked immunosorbentassay,ELISA)法测定细胞上清液中白介素18(interleukin 18,IL-18)和白介素-1β(interleukin 1 beta,IL-1β)表达水平;采用Western blot检测GasderminD蛋白(GSDMD-N)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、白介素1β前体(pro-interleukin 1,pro-IL-1β)、半胱氨酸天冬氨酸蛋白酶1(caspase-1)、Nrf2以及凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)表达情况;采用细胞乳酸脱氢酶(lactate dehydrogenase,LDH)试剂盒法测定细胞内LDH表达变化情况,采用Hochest/PI染色法观察细胞凋亡情况。结果与对照组比较,OGD/R组细胞存活率[(40.16±3.39)%比(101.3±1.54)%]、Nrf2表达水平[(0.47±0.09)比(1.00±0.00)]明显降低;IL-1β[(46.55±9.59)pg/mL比(9.35±3.33)pg/mL]、IL-18[(35.39±10.63)pg/mL比(7.85±3.04)pg/mL]、LDH渗出率[(0.36±0.04)%比(0.18±0.08)%]、GSDMD-N[(2.24±0.36)比(1.00±0.00)]、NLRP3[(2.15±0.23)比(1.00±0.00)]、pro-IL-1β[(3.08±0.17)比(1.00±0.00)]、caspase-1[(2.20±0.32)比(1.00±0.00)]表达水平明显升高,差异有统计学意义(P<0.001);与OGD/R组比较,醒脑静注射液组细胞存活率[(63.69±11.28)%比(40.16±3.39)%]、Nrf2表达水平[(0.89±0.11)比(0.47±0.09)]明显升高,LDH渗出率[(0.25±0.07)%比(0.36±0.04)%]、IL-1β[(33.30±8.60)pg/mL比(46.55±9.59)pg/mL]、IL-18[(21.72±7.50)pg/mL比(35.39±10.63)pg/mL]、GSDMD-N[(1.34±0.06)比(2.24±0.36)]、NLRP3[(1.25±0.18)比(2.15±0.23)]、pro-IL-1β[(1.65±0.30)比(3.08±0.17)]、caspase-1[(1.23±0.15)比(2.20±0.32)]表达水平明显降低,差异有统计学意义(P<0.05);与醒脑静组比较,醒脑静+Nrf2抑制剂组Nrf2蛋白表达水平[(0.38±0.12)比(0.89±0.11)]明显降低,LDH渗出率[(0.36±0.07)%比(0.25±0.07)%]、IL-1β[(53.28±6.37)pg/mL比(33.30±8.60)pg/mL]、IL-18[(49.56±8.87)pg/mL比(21.72±7.50)pg/mL]、GSDMD-N[(1.75±0.27)比(1.34±0.06)]、NLRP3[(2.22±0.17)比(1.25±0.18)]、pro-IL-1β[(3.54±0.50)比(1.65±0.30)]、caspase-1[(2.36±0.28)比(1.23±0.15)]蛋白表达水平明显升高,差异有统计学意义(P<0.05)。结论Nrf2抑制剂可逆转醒脑静注射液对OGD/R损伤诱导SH-SY5Y细胞焦亡的保护作用;醒脑静注射液通过上调Nrf2蛋白表达,抑制OGD/R损伤诱导的SH-SY5Y细胞焦亡。展开更多
基金The study is financially supported by the Development Foundation ofScience and Technology under Shandong Provincial Education Department(No. J05D51).
文摘p-nitrotoluene is an important organic intermediate widely used in pesticide, foamed plastics, dyestuff and medicine industries. In this paper, Crusian carps (Carassius auratus) were exposed to dilute p-nitrotoluene solutions with different concentrations of 0, 20, 40, 80, 120, 160, 240 and 320 μg/L; the subsequent physiological responses to the chemical were observed, and the variation characteristics of blood corpuscles, ovaries and oocytes were studied by bio-microscopy. Results showed that even very low concentrations of p-nitrotoluene would lead to toxicosis and even the death of Carassius auratus. The bio-microscopic structures revealed that, compared with the compartment of Carassius auratus growing in the p-nitrotoluene–free water (the control group), the lower concentrations of p-nitrotoluene (for example no more than 80 μg/L) would result in the faster disintegration of erythrocytes and the significant enlargement of oocytes, while the higher contents of p-nitrotoluene (no less than 240 μg/L in this case) would inhibit the division of cells and their further growth, even degenerate blood corpuscles and ovaries, suggesting stronger estrogenic activities.
文摘目的探讨醒脑静注射液对缺糖缺氧再灌注(oxygen and glucose deprivation/reoxygenation,OGD/R)损伤诱导SH-SY5Y细胞焦亡的保护作用及机制。方法使用不同浓度醒脑静注射液预处理SH-SY5Y细胞,确定醒脑静注射液给药浓度;将SH-SY5Y细胞随机分为对照组、OGD/R组、醒脑静注射液+OGD/R组(10μL/mL醒脑静注射液)以及醒脑静注射液+OGD/R+核因子E2相关因子2(nuclear factor-E2 related factor 2,Nrf2)抑制剂组(10μL/mL醒脑静注射液+2μmol/mL ML385)共4组,每组实验进行6次(n=6),采用MTT比色法检测细胞存活率;采用酶联免疫吸附试验(enzyme-linked immunosorbentassay,ELISA)法测定细胞上清液中白介素18(interleukin 18,IL-18)和白介素-1β(interleukin 1 beta,IL-1β)表达水平;采用Western blot检测GasderminD蛋白(GSDMD-N)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、白介素1β前体(pro-interleukin 1,pro-IL-1β)、半胱氨酸天冬氨酸蛋白酶1(caspase-1)、Nrf2以及凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)表达情况;采用细胞乳酸脱氢酶(lactate dehydrogenase,LDH)试剂盒法测定细胞内LDH表达变化情况,采用Hochest/PI染色法观察细胞凋亡情况。结果与对照组比较,OGD/R组细胞存活率[(40.16±3.39)%比(101.3±1.54)%]、Nrf2表达水平[(0.47±0.09)比(1.00±0.00)]明显降低;IL-1β[(46.55±9.59)pg/mL比(9.35±3.33)pg/mL]、IL-18[(35.39±10.63)pg/mL比(7.85±3.04)pg/mL]、LDH渗出率[(0.36±0.04)%比(0.18±0.08)%]、GSDMD-N[(2.24±0.36)比(1.00±0.00)]、NLRP3[(2.15±0.23)比(1.00±0.00)]、pro-IL-1β[(3.08±0.17)比(1.00±0.00)]、caspase-1[(2.20±0.32)比(1.00±0.00)]表达水平明显升高,差异有统计学意义(P<0.001);与OGD/R组比较,醒脑静注射液组细胞存活率[(63.69±11.28)%比(40.16±3.39)%]、Nrf2表达水平[(0.89±0.11)比(0.47±0.09)]明显升高,LDH渗出率[(0.25±0.07)%比(0.36±0.04)%]、IL-1β[(33.30±8.60)pg/mL比(46.55±9.59)pg/mL]、IL-18[(21.72±7.50)pg/mL比(35.39±10.63)pg/mL]、GSDMD-N[(1.34±0.06)比(2.24±0.36)]、NLRP3[(1.25±0.18)比(2.15±0.23)]、pro-IL-1β[(1.65±0.30)比(3.08±0.17)]、caspase-1[(1.23±0.15)比(2.20±0.32)]表达水平明显降低,差异有统计学意义(P<0.05);与醒脑静组比较,醒脑静+Nrf2抑制剂组Nrf2蛋白表达水平[(0.38±0.12)比(0.89±0.11)]明显降低,LDH渗出率[(0.36±0.07)%比(0.25±0.07)%]、IL-1β[(53.28±6.37)pg/mL比(33.30±8.60)pg/mL]、IL-18[(49.56±8.87)pg/mL比(21.72±7.50)pg/mL]、GSDMD-N[(1.75±0.27)比(1.34±0.06)]、NLRP3[(2.22±0.17)比(1.25±0.18)]、pro-IL-1β[(3.54±0.50)比(1.65±0.30)]、caspase-1[(2.36±0.28)比(1.23±0.15)]蛋白表达水平明显升高,差异有统计学意义(P<0.05)。结论Nrf2抑制剂可逆转醒脑静注射液对OGD/R损伤诱导SH-SY5Y细胞焦亡的保护作用;醒脑静注射液通过上调Nrf2蛋白表达,抑制OGD/R损伤诱导的SH-SY5Y细胞焦亡。
