刘立新 高春艳 古同男 任丽丽 鄢雯.H2O2诱导小鼠肌原细胞系C2C12 UII/UT 系统的高表达[J].现代生物医学进展英文版,2015,15(30):5830-5833. |
H2O2诱导小鼠肌原细胞系C2C12 UII/UT 系统的高表达 |
H2O2 Induced the Up-regulation of UII/UT Systemin Mouse Myogenic Cell Line C2C12 |
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DOI: |
中文关键词: 尾加压素II G 蛋白偶联受体 过氧化氢 |
英文关键词: Urotensin II G protein coupled receptor Hydrogen peroxide |
基金项目:北京高等学校青年英才项目(YETP1679);首都医科大学校自然科学基金项目(2014QD03) |
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中文摘要: |
目的:观察外源给予H2O2对C2C12 细胞中UII/UT 系统表达的影响。方法:培养小鼠肌原细胞系C2C12 细胞株,应用胎盘蓝
染色和测定细胞培养液中LDH的含量检测H2O2对细胞损伤的影响,应用放射免疫的方法检测细胞培养液和裂解液中UII的含
量,应用RT-PCR法测定C2C12 中UT mRNA 的表达,应用Western Blot 检测不同浓度H2O2对肌原细胞系C2C12 中UT 蛋白表
达的影响。结果:外源给予不同浓度的H2O2,C2C12细胞裂解液中UII的含量各组间无明显差异,但增加了细胞孵育液中UII的含
量,10-4M 和10-3M 时分别增加了83.1 %(p<0.01)和94.5 %(P<0.01)。低浓度H2O2(0.5× 10-6M, 10-6M, 10-5M, 10-4M)刺激明显增
加了UT mRNA 的表达,而高浓度的H2O2(10-3M)刺激反而使UT mRNA 的表达降低。高浓度的H2O2(10-6 M, 10-5M, 10-4M, 10-3
M)刺激使UT 蛋白表达分别增加了200.1 %、255.6 %、111.1 %和100.1 %(均p<0.05)。结论:H2O2作为T2DM 发病因素之一的活
性氧族,可能是T2DM 时骨骼肌组织中UII/UT 系统表达增加的一个因素。 |
英文摘要: |
Objective:To observe the effect of H2O2 on the expression of UII/UT system in C2C12.Methods:Cell injury was
determined by Placenta blue staining and LDH kit, the content of UII in cell lysate and cell incubation fluid were assayed by RIA, mRNA
levels were assayed by RT-PCR, Protein levels were assayed by Western Blot.Results:UII content in cell incubation fluid was increased
after H2O2 of different concentration was administrated, the UII content was increased by 83.1 %(P<0.01) and 94.5 %(P<0.01) when the
concentration of H2O2 was elevated to 10-4Mand 10-3 M. The simulation of different concentration of H2O2 (0.5× 10-6 M, 10-6M, 10-5 M
and 10-4 M) significantly increased the expression of UT mRNA. However, The expression of UT mRNA was decreased significantly
when the concentration of H2O2 was elevated to 10-3 M. The UT protein expression was increased by 200.1%, 200.1%, 255.6% and
200.1%when the concentration of H2O2 was elevated to 10-6M, 10-5M, 10-4Mand 10-3M (P<0.05).Conclusion:H2O2might be a factor
involved in up-regulating the expression of UII/UT systemfor the skeletal muscle of Type 2 diabetes mellitus. |
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