CAJ | 학술논문

目的评价乌司他丁(UTI)对心肺复苏(CPR)后大鼠大脑皮层炎症因子TNF-α和IL-6表达及神经元细胞凋亡的影响.方法 36只成年雄性Wistar大鼠采用交流电致颤的方式诱发室颤(VF),持续7 min后进行CPR,建立VF/CPR模型.自主循环恢复(ROSC)后立即给予乌司他丁100 000 U/kg或等体积的PBS静脉注射.VF前和ROSC后2、4、8h采血检测血浆TNF-α、IL-6水平,并取大脑皮层进行定量PCR和Western blot检测TNF-α、IL-6 mRNA表达和蛋白表达,检测核因子NF-κB p65的核浆转位情况.ROSC后72 h行皮层NISSL和TUNEL染色计数顶叶皮层存活神经元细胞数和凋亡神经元细胞数.结果 ROSC后UTI组2、4、8h血浆TNF-α质量浓度分别为(17.7±1.4)、(21.9±2.1)和(17.1±0.6),低于PBS组(t2h=1.42,t4h =2.93,t8 h =4.22,各组P＜0.05).UTI组2、4、8h血浆IL-6质量浓度(ng/mL)分别为(208.9±14.1)、(281.5 ±25.9)和(251.8±15.3),显著低于PBS组(t2h=3.87,t4h=2.45,t8h=3.74,各组P＜0.05).ROSC后UTI组2、4、8h大脑皮层TNF-α、IL-6 mRNA表达和蛋白表达量均显著低于PBS组.PBS组ROSC后2、4、8h大脑皮层NF-κB p65的核/浆转换率为(1.08±0.08)、(1.02±0.05)和(0.97 ±0.02),高于UTI组.ROSC后72 h UTI组大脑皮层存活神经元细胞数为(22±3)个/400×400像素,多于PBS组的(19±2)个/400×400像素(Z=2.887,P=0.02);凋亡细胞数UTI组为(10±2)个/400×400像素,低于PBS组的(13±3)个/400×400像素(Z =3.751 P=0.01).结论 UTI降低ROSC后Wistar大鼠全身炎症反应,减少大脑皮层NF-κB通路的激活,减少致炎因子TNF-α和IL-6的表达,从而减少神经元细胞凋亡,促进细胞存活.
목적 평개오사타정(UTI)대심폐복소(CPR)후대서대뇌피층염증인자TNF-α화IL-6표체급신경원세포조망적영향.방법 36지성년웅성Wistar대서채용교류전치전적방식유발실전(VF),지속7 min후진행CPR,건립VF/CPR모형.자주순배회복(ROSC)후립즉급여오사타정100 000 U/kg혹등체적적PBS정맥주사.VF전화ROSC후2、4、8h채혈검측혈장TNF-α、IL-6수평,병취대뇌피층진행정량PCR화Western blot검측TNF-α、IL-6 mRNA표체화단백표체,검측핵인자NF-κB p65적핵장전위정황.ROSC후72 h행피층NISSL화TUNEL염색계수정협피층존활신경원세포수화조망신경원세포수.결과 ROSC후UTI조2、4、8h혈장TNF-α질량농도분별위(17.7±1.4)、(21.9±2.1)화(17.1±0.6),저우PBS조(t2h=1.42,t4h =2.93,t8 h =4.22,각조P＜0.05).UTI조2、4、8h혈장IL-6질량농도(ng/mL)분별위(208.9±14.1)、(281.5 ±25.9)화(251.8±15.3),현저저우PBS조(t2h=3.87,t4h=2.45,t8h=3.74,각조P＜0.05).ROSC후UTI조2、4、8h대뇌피층TNF-α、IL-6 mRNA표체화단백표체량균현저저우PBS조.PBS조ROSC후2、4、8h대뇌피층NF-κB p65적핵/장전환솔위(1.08±0.08)、(1.02±0.05)화(0.97 ±0.02),고우UTI조.ROSC후72 h UTI조대뇌피층존활신경원세포수위(22±3)개/400×400상소,다우PBS조적(19±2)개/400×400상소(Z=2.887,P=0.02);조망세포수UTI조위(10±2)개/400×400상소,저우PBS조적(13±3)개/400×400상소(Z =3.751 P=0.01).결론 UTI강저ROSC후Wistar대서전신염증반응,감소대뇌피층NF-κB통로적격활,감소치염인자TNF-α화IL-6적표체,종이감소신경원세포조망,촉진세포존활.
Objective To evaluate the effects of Ulinastatin (UTI) on the expressions of TNF-α,IL-6 and neurons apoptosis in cerebral cortex of rats after cardiopulmonary resuscitation (CPR).Methods Thirty-six healthy male adult Wistar rats were induced ventricular fibrillation untreated for 7 min and then received CPR.The animals were infused UTI 100 000 U/kg or phosphate-buffered solution (PBS) at once after ROSC.At 2,4 and 8 h after ROSC,cerebral cortex were removed to determine the mRNA expressions and levels of TNF-α protein and IL-6 protein,the translocation ratio of NF-κB p65 from cytoplasm to nucleus and the apoptotic neurons.Results The plasma levels of TNF-α (ng/mL) in animals of UTI group were (17.7 ± 1.4),(21.9 ± 2.1) and (17.1 ± 0.6) at 2,4 and 8 h after ROSC respectively,and significantly lower than those in PBS group at the given intervals.Mean while,the levels of IL-6 (ng/mL) were (208.9 ± 14.1),(281.5 ±25.9) and (251.8 ± 15.3) at 2,4 and 8 h after ROSC respectivèly in animals of UTI group,and lower than those in PBS group.The expressions of TNF-α mRNA and IL-6 mRNA and protein levels of TNF-α and IL-6 in UTI group were both lower than those in PBS group at given intervals,respectively.The translocation ratio of NF-κB p65 from plasma to nucleus in PBS group at each given interval after ROSC was significantly higher than that in UTI group.The number of viable neurons in cerebral cortex in UTI group was higher than that in PBS group,while the number apoptosis neurons was fewer in UTI group.Conclusions UTI attenuated the general inflammatory response after ROSC in rat,decreased the activation of NF-κB pathway,and subsequently attenuated the expression of TNF-α and IL-6,and finally decreased the neurons apoptosis.