摘要
目的研究甲基胍对人肾小管上皮细胞凋亡的影响,以及前列腺素E1(PGE1)和普罗布考的抗凋亡作用。方法HK-2细胞分5组培养,A正常对照组;B肌酐组:培养基中加入0.5 mmol.L-1肌酐;C甲基胍组:培养基中加入0.5 mmol.L-1甲基胍;D PGE1组:培养基中加入0.5 mmol.L-1甲基胍和2μg.L-1PGE1;E普罗布考组:培养基中加入0.5 mmol.L-1甲基胍和20μmol.L-1普罗布考。培养48 h后用MTT比色法测OD值,并检测培养液NAG酶(N-乙酰-β-氨基葡萄糖苷酶)活性;用Hoechst染色及Annexin V-FITC/PI流式细胞仪检测细胞凋亡。结果①肌酐组和甲基胍组HK-2细胞OD值较正常对照组显著下降(P<0.01),NAG酶活性上升(P<0.01);甲基胍组与肌酐组比较,OD值和酶活性有显著性差异(P<0.01);PGE1组和普罗布考组与甲基胍组和肌酐组比较,OD值显著上升而NAG酶活性下降,PGE1改变幅度大于普罗布考组(P<0.01)。②HK-2细胞加入甲基胍后,Hoechst33258染色显示凋亡细胞呈现核固缩,染色加深,并出现凋亡小体。③流式细胞显示甲基胍组HK-2细胞凋亡率显著高于对照组和肌酐组(P<0.01)。PGE1和普罗布考组的凋亡率显著下降(P<0.01),PGE1组下降幅度大于普罗布考组(P<0.05)。结论肌酐、甲基胍有促进肾小管上皮细胞凋亡的作用,后者较前者明显。PGE1、普罗布考对甲基胍所致肾小管上皮细胞凋亡有一定的保护作用。
Objective To investigate the apoptosis in methylguanidine induced human renal tubular epithelial ceil line （HK-2） and the antiapoptosis effect of PGE1 and probucol. Methods （1）HK-2 was divided into 5 groups to culture： a normal control group （A）, a ereatinine group （B） with 0.5 mmol · L ^-1 creatinine in the medium, a methylguanidine group （C） with 0.5 mmol · L ^-1methylguanidine in the medium, a PGE1 group （D） with 0.5 mmol · L ^-1 mthylguanidine and 2 fig · L ^-1 PGE1, and a probucol group （E） with 0.5 mmol · L ^-1 methylguanidine and 20 μmol · L ^-1probueol in the medium. （2） OD value of HK 2 was detected by MTT and NAG concentration was determined. （3） Cell apoptosis was evaluated by Hoeehst stain and FACS with Annexin-V/PI. Resulls OD value and NAG concentration were significantly different in the 5 groups, B and C group compared with that of A group （P〈 0.01 ）, and the difference was more obvious between C and B group （P〈0.01 ）. OD value decreased and NAG concentration increased and the difference was significant （P〈0.05） Between D and E group, By Hoechst stain, HK-2 cells showed that pycnosis, apoptotic body and deepened stain of cells could be found when HK-2 was cultivated with methylguanidine. The apoptotic rate of HK 2 increased more significantly in C group than other groups （P〈0.01）. The apoptotie rate of D and E group significantly decreased. Significant difference was shown between D and E group （P〈0.05）. Conclusion Creatinine and methylguanidine can promote the apoptosis of renal tubular epithelial cells and the former is more significant. PGE1 and probucol can protect renal tubular epithelial cells from methylguanidine induced apoptosis.
出处
《中南药学》
CAS
2010年第1期20-23,共4页
Central South Pharmacy
关键词
甲基胍
肾小管上皮细胞
凋亡
尿毒症
methylguanidine
renal tubular epithelial cell
apoptosis
urinemia
