摘要
目的探讨内毒素在体外作用于大鼠肾髓质上皮细胞mIMCD-3的作用。方法用终浓度为0.1、0.5、1.0、5.0和10.0μg/L的内毒素与大鼠肾髓质上皮细胞mIMCD-3培养12、24、48和72h后用CCK-8检测吸光度,计算细胞抑制率。选取作用明显的终浓度为10μg/L的内毒素作用于mlMCD-3后分别在4、12、24和48h用流式细胞仪检测凋亡。结果 LPS抑制肾髓质上皮细胞mIMCD-3增殖。不同浓度间分析,差异具有统计学意义(P<0.05)。当浓度增大到1μg/L及以上时,LPS对细胞的抑制作用明显增强,具有浓度依赖性。不同时间组分别行单因素方差比较,差异具有统计学意义(P<0.05)。浓度为5μg/L及以上时,任意两两组间对比均差异具有统计学意义,具有时间依赖性。LPS诱导肾髓质上皮细胞mIMCD-3凋亡。经流式细胞仪检测,结果表明,LPS对mIMCD-3的抑制作用主要表现为细胞凋亡,随作用时间的延长,细胞凋亡增加(P<0.01),同时作用时间和凋亡率具有直线相关性(R=0.9920)。结论内毒素能在体外诱导大鼠肾髓质上皮细胞mIMCD-3凋亡抑制增殖。
[Objective] To research the effect of LPS on rat kidney medulla epithelial cells （mIMCD-3）. [ Methods ] mIMCD-3 cells were seeded in 96-well plates and incubated for 0-72 h with different concentration of endotoxin. The CCK-8 was used to determine the inhibitory effect. Selecting the final concentration of 10μmol/L of endotoxin to be euhured with cells for 0-48 h, flow eytometry was used to calculate inhibitory rate. [Results] mIMCD-3 expressed AQP2. After immunohistoehemieal staining, a large number of brown-red particles piled up like plates could be seen in the cytoplasm. It proved that APQ2 localized in the cytoplasm in the basic state. At the same time, negative control cells which didn＇t add first antibody, showed none of brown-red particles generated in the cytoplasm. The entire cytoplasm was stained light blue. Endotoxin inhibited mIMCD-3 proliferation. One-way ANOVA was used to analyze the significance of concentration, the results had statistie significance （P 〈0.05）. Doing inter-group analysis had proved that when the concentration was less than 5μg/L, the inhibition of cell proliferation was not very apparent. When the concentration gradient increased, the inhibitory effect increased with dosedependent manner. With the concentration of 0.1μg/L the cells did not inhibit, but proliferated. When the time was up to 72 hours, the adjacent inter-group comparison had non-statistical differences. Groups of different times were compared with one-way ANOVA, it had statistical difference （P 〈0.05）. However, it proved that when the concentration was less than 1μg/L, the inhibitory rate changed little over time. Once greater than or equal to the concentration of 5μg/L, the contrasts between any two groups were significantly different. It showed a timedependent manner. Endotoxin induced apoptosis. Flow cytometry results showed that the inhibition of LPS on mIMCD-3 was mainly expressed in the manner of apoptosis. When time extended, apoptosis rate increased （P 〈0.01） and the rate of apop
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第13期1971-1974,共4页
China Journal of Modern Medicine
