摘要
目的观察阿魏酸钠(sodium ferulate,SF)对高糖培养的人近端肾小管上皮细胞(human proximal tubular epithelial cells,HKC)增殖和凋亡的影响。方法选用对数期生长的肾小管上皮细胞,分为对照组、高糖组、高糖加阿魏酸钠组。四甲基偶氮唑盐(microculture tetrazolium,MTT)法观察细胞增殖能力的变化,测定细胞培养液中的乳酸脱氢酶(lactate dehydrogenase,LDH)的含量以判断细胞损伤情况,Hoechst 33258荧光染色检测细胞凋亡。结果与对照组比较,高糖组MTT吸光度值(OD值)随培养时间延长明显降低,而高糖加阿魏酸钠组OD值与高糖组比较有明显升高(P<0.05)。随培养时间的延长,高糖组培养基中LDH水平明显升高,在12 h和24 h分别为(17.55±2.45)和(28.72±3.11)U/L,而高糖加阿魏酸钠组12 h和24 h LDH水平降至(11.84±2.18)和(19.98±3.67)U/L,和同时间点高糖组比较均有明显降低(P<0.05)。高糖处理组培养液中的丙二醛(malondiadehycle,MDA)水平较对照组明显增加(P<0.01),超氧化物歧化酶(superoxidedismutase,SOD)活性明显降低(P<0.01),阿魏酸钠组培养液中MDA水平明显降低(P<0.01),SOD活性明显增加(P<0.05,P<0.01)。Hoechst 33258荧光染色显示,对照组和高糖加阿魏酸钠组仅见个别凋亡形态细胞,高糖组典型凋亡形态细胞明显增多,细胞凋亡率为34.17%,显著高于正常对照组(5.20%)和高糖加阿魏酸钠组(7.39%,P<0.01)。结论高糖培养可诱导人近端肾小管上皮细胞的损伤和凋亡,并抑制其增殖。阿魏酸钠对人肾小管上皮细胞系(human kidney cell,HKC)的保护作用可能是通过抑制HKC的损伤和凋亡,以及提高增殖能力实现的。
Objective To observe the influence of sodium ferulate （SF） on proliferation and apoptosis of human proximal tubular epithelial cells （HKCs） induced by high glucose. Methods The renal tubular epithelial cells in logarith- mic growth phase were divided into control group, high glucose group and high glucose plus SF group. The cell prolifera- tion ability was measured by MTT assay. The culture solution concentrations of lactate dehydrogenase （LDH） were measured to evaluate cell injury. The culture solution of malondiadehycle （MDA） levels and superoxidedismutase （SOD） activities were measured to assess oxidative stress, and cell apoptosis were detected with Hoechst 33258. Results MTF absorbance （OD value） of high glucose group was decreased significantly along with the culture time compared with control group. The OD value of high glucose plus SF group was reduced significantly compared with high glucose group （P〈0. 05）. With the prolongation of culture time, the culture solution concentration of LDH was increased significantly and were （17. 55 ± 2. 45） and （28. 72±3. 11） U/L at 12 and 24 hours respectively. LDH levels were reduced to （11.84 ±2. 18） and （19. 98 ±3. 67） U/L at 12 and 24 hours in high glucose plus SF group （P〈0. 05）. The culture solution of MDA levels were higher （P〈 0. 01）, the SOD activities were significantly reduced （P〈0. 01） in high glucose group than that in control group. The MDA lev- els were significantly reduced （P%0. 01） and SOD activities were significantly increased （P〈0. 05 or 0. 01） in high glucose plus SF group. Hoechst 33258 fluorescent staining revealed that the nucleus crimpled, crescent liked and chromatin condensed, even disintegrated. The apoptosis rate of high glucose group （34. 17%） was higher than that in the control group （5.20%） and high glucose plus SF group （7.39%, P〈 0. 01）. Conclusion High glucose can induce HKC injury and apoptosis, and inhibit its proliferation. The protective of SF may be correlated with its effects to inhibit damage and apoptosis, and improve the proliferation ability of HKC.
出处
《华南国防医学杂志》
CAS
2013年第6期379-382,共4页
Military Medical Journal of South China
基金
解放军总医院科技创新苗圃基金(10KMM26)
关键词
阿魏酸钠
高糖
人近端肾小管上皮细胞
Sodium ferulate
High glucose
Human proximal tubular epithelial cell
