CAJ | 학술논문

目的评价不同浓度PNU282987对大鼠心肌细胞缺氧/复氧损伤的影响.方法新生大鼠,分离纯化心肌细胞,培养72 h时,以5×105个/ml的密度接种于6孔板中(2 ml/孔),108个培养孔,采用随机数字表法,分为6组(n=18):正常对照组(C组)、缺氧/复氧组(A/R组)、3、10、30、60μmol/L PNU282987组(P3组、P10组、P30组和P60组).采用缺氧6h复氧6h的方法制备心肌细胞缺氧/复氧损伤模型,P3组、P10组、P30组和P60组复氧时分别加入相应浓度的PNU282987.采用比色法检测心肌细胞乳酸脱氢酶(LDH)漏出水平,采用膜联蛋白V/碘化丙啶双染流式细胞术检测心肌细胞凋亡情况,采用ELISA法检测培养上清液白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的浓度.结果与C组比较,A/R组、P3组、P10组、P30组和P60组心肌细胞LDH漏出率、早期细胞凋亡率和晚期细胞凋亡率升高,培养上清液IL-6和TNF-α的浓度升高(P＜0.01);与A/R组比较,P3组、P10组、P30组和P60组心肌细胞LDH漏出率和早期细胞凋亡率降低,P30组最明显,培养上清液IL-6和TNF-α的浓度降低,且呈浓度依赖性(P＜0.05或0.01),P3组、P10组、P30组和P60组晚期细胞凋亡率差异无统计学意义(P＞0.05).结论 3、10、30、60 μmol/L PNU282987可减轻大鼠心肌细胞缺氧/复氧损伤,而30μmol/L时效果最明显.
목적 평개불동농도PNU282987대대서심기세포결양/복양손상적영향.방법 신생대서,분리순화심기세포,배양72 h시,이5×105개/ml적밀도접충우6공판중(2 ml/공),108개배양공,채용수궤수자표법,분위6조(n=18):정상대조조(C조)、결양/복양조(A/R조)、3、10、30、60μmol/L PNU282987조(P3조、P10조、P30조화P60조).채용결양6h복양6h적방법제비심기세포결양/복양손상모형,P3조、P10조、P30조화P60조복양시분별가입상응농도적PNU282987.채용비색법검측심기세포유산탈경매(LDH)루출수평,채용막련단백V/전화병정쌍염류식세포술검측심기세포조망정황,채용ELISA법검측배양상청액백세포개소-6(IL-6)화종류배사인자-α(TNF-α)적농도.결과 여C조비교,A/R조、P3조、P10조、P30조화P60조심기세포LDH루출솔、조기세포조망솔화만기세포조망솔승고,배양상청액IL-6화TNF-α적농도승고(P＜0.01);여A/R조비교,P3조、P10조、P30조화P60조심기세포LDH루출솔화조기세포조망솔강저,P30조최명현,배양상청액IL-6화TNF-α적농도강저,차정농도의뢰성(P＜0.05혹0.01),P3조、P10조、P30조화P60조만기세포조망솔차이무통계학의의(P＞0.05).결론 3、10、30、60 μmol/L PNU282987가감경대서심기세포결양/복양손상,이30μmol/L시효과최명현.
Objective To evaluate the effects of different concentrations of PNU282987 on anoxia/ reoxygenation (A/R) injury to cardiomyocytes of rats.Methods After being cultured for 72 h, cardiomyocytes obtained from neonatal rats were seeded in 6-well plates (2 ml/well, a total of 108 wells) at a density of 5× 105 cells/ml and randomly divided into 6 groups according to a random number table (n = 18 each) : control group (C group);A/R group;3, 10, 30 and 60 μmol/L PNU282987 groups (P3,P10, P30 and P50 groups).The cells were exposed to anoxia for 6 h followed by reoxygenation with 95％ O2-5％ CO2at 37 ℃ for 6 h.The corresponding concentrations of PNU282987 were added to the culture medium during reoxygenation in P3, P10, P30 and P60 groups.The amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.Apoptosis in cardiomyocytes was detected by flow cytometry with Annexin V/propidium iodide staining.The concentrations of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the supernatant were determined using enzyme-linked immunosorbent assay.Results Compared to C group, the LDH release rate, early and late apoptosis rates, and the concentrations of IL-6 and TNF-α in the supernatant were significantly increased in A/R, P3, P10, P30 and P60 groups.Compared to A/R group, the LDH release rate and early apoptosis rate were significantly decreased (especially in group P30), and the concentrations of IL-6 and TNF-α in the supernatant were decreased in a concentration-dependent manner in P3, P10, P30 and P60 groups.Compared to A/R group, no significant changes were found in late apoptosis rate in P3, P10, P30 and P60 groups.Conclusion PNU282987 3, 10, 30 and 60 μmol/L can reduce A/R injury to cardiomyocytes of rats, especially 30 μmol/L.