CAJ | 학술논문

目的将布比卡因作用于高糖培养的SH-SY5Y细胞,观测其对细胞内活性氧簇(reactive oxygen species,ROS)及细胞凋亡的影响. 方法不同浓度葡萄糖培养基培养SH-SY5Y细胞,经1 mmol/L布比卡因作用6h.采用流式细胞仪检测细胞内ROS产量及细胞凋亡情况、MTT法检测细胞存活率、Western blot技术检测细胞内GRP78相关蛋白的变化. 结果葡萄糖浓度为5.6、6.1、7.0 mmol/L的3组间ROS含量差异无统计学意义(P＞0.05);葡萄糖浓度为7.8、11.1、13.3 mmol/L时ROS产量分别高于5.6、6.1、7.0 mmol/L组(P＜0.05).各组细胞存活率(％)依次为(4.29±0.20)、(3.30±0.19)、(6.31±0.14)、(7.89±0.09)、(2.81±0.12、(1.01±0.11,组间差异均具有统计学意义(P＜0.05).各组细胞凋亡率(％)依次为(1.80±0.09)、(2.82±0.11)、(3.30±0.21)、(431±0.18)、(6.28±0.13)、(7.89型0.08),组间差异均具有统计学意义(P＜0.05).Grp78表达结果为5.6 mmol/L组和6.1 mmol/L组分别为(1.02±0.12)和(0.97±0.06),均高于7.0 mmol/L组(0.39±0.04);7.8 mmol/L组(0.98±0.06)高于7.0 mmol/L组(0.39±0.04);11.1 mmol/L与13.3 mmol/L组低于其他各组,差异均具有统计学意义(P＜0.05);5.6 mmol/L组、6.1 mmol/L组和7.8 mmol/L组相比,差异无统计学意义(P＞0.05). 结论细胞内ROS增多及细胞凋亡增加可能与布比卡因增强高糖培养的SH-SY5Y细胞内质网应激有关,引起细胞内ROS爆发,从而导致细胞损伤和凋亡;而Grp78功能减弱进一步增强细胞内质网应激.
목적 장포비잡인작용우고당배양적SH-SY5Y세포,관측기대세포내활성양족(reactive oxygen species,ROS)급세포조망적영향. 방법 불동농도포도당배양기배양SH-SY5Y세포,경1 mmol/L포비잡인작용6h.채용류식세포의검측세포내ROS산량급세포조망정황、MTT법검측세포존활솔、Western blot기술검측세포내GRP78상관단백적변화. 결과 포도당농도위5.6、6.1、7.0 mmol/L적3조간ROS함량차이무통계학의의(P＞0.05);포도당농도위7.8、11.1、13.3 mmol/L시ROS산량분별고우5.6、6.1、7.0 mmol/L조(P＜0.05).각조세포존활솔(％)의차위(4.29±0.20)、(3.30±0.19)、(6.31±0.14)、(7.89±0.09)、(2.81±0.12、(1.01±0.11,조간차이균구유통계학의의(P＜0.05).각조세포조망솔(％)의차위(1.80±0.09)、(2.82±0.11)、(3.30±0.21)、(431±0.18)、(6.28±0.13)、(7.89형0.08),조간차이균구유통계학의의(P＜0.05).Grp78표체결과위5.6 mmol/L조화6.1 mmol/L조분별위(1.02±0.12)화(0.97±0.06),균고우7.0 mmol/L조(0.39±0.04);7.8 mmol/L조(0.98±0.06)고우7.0 mmol/L조(0.39±0.04);11.1 mmol/L여13.3 mmol/L조저우기타각조,차이균구유통계학의의(P＜0.05);5.6 mmol/L조、6.1 mmol/L조화7.8 mmol/L조상비,차이무통계학의의(P＞0.05). 결론 세포내ROS증다급세포조망증가가능여포비잡인증강고당배양적SH-SY5Y세포내질망응격유관,인기세포내ROS폭발,종이도치세포손상화조망;이Grp78공능감약진일보증강세포내질망응격.
Objective To discuss bupivacaine causing SH-SY5Y cells in high sugar environment to increase reactive oxygen species (ROS) and apoptosis.Methods we adopt 1 mmol/L bupivicaine to SH-SY5Y cells cultured by different concentrations of glucose medium,and employ flow cytometry to detect the quantity of ROS in the cells and the situation of apoptosis.we use MTT method to detect cell survival rate.At last we use western blotting technique to detect the change of the related protein about GRP78.Results Groups of cells containing different concentration of sugar medium through bupivacaine impacted 6 h have been detected that the contents of intarcellular ROS in groups of 5.6,6.1,7 mmol/L have no statistical significance (P＞0.05).Each group of cell survival rate (％) in sequence is (4.29±0.20),(3.30±0.19),6.31±0.14),(7.89±0.09),(2.81±0.12),(1.01±0.11),and cell survival rate among groups have statistical significance(P＜0.05).Each group of cell apoptosis rate(％) in sequence is (1.80±0.09),(2.82± 0.11),(3.30±0.21),(4.31±0.18),(6.28±0.13),(7.89±0.08),and cell apoptosis rate among groups have statistical significance (P＜ 0.05).Western blot detect the expression of Grp78 in group of 5.6 mmol/L and 6.1 mmol/L are (1.02±0.12) and (0.97±0.06) respectively,exceeding the group of 7.0 mmol/L.Also the difference have statistical significance (P＜0.05).The groups of 11.1 mmol/L and 13.3 mmol/L being shorter than the others and compared to the others have statistical significance (P＜0.05).Conclusions Intracellular ROS increase and apoptosis are associated with bupivacaine that further strengthen endoplasmic reticulum stress.Endoplasmic reticulum stress cause ROS to outbreak in cells,leading cells to damage and apoptos.With the function of GRP78 degrading,endoplasmic reticulum stress strengthen than ever.