CAJ | 학술논문

[目的]观察环孢素A(CSA)对近端肾小管上皮细胞基质金属蛋白酶-2和-9(MMP-2和MMP-9)表达和活性的影响.[方法]体外培养大鼠近端肾小管上皮细胞株NRK52E细胞至完全融合时,分别给予不同浓度的CSA 0,0.42,0.84,4.2,8.4 μmoL/L,实验观察48 h和72 h.收集培养液,用明胶酶谱法检测MMP-2和MMP-9活性.提取总RNA,逆转录聚合酶链式反应(RT-PCR)检测MMP-2和MMP-9的mRNA水平.[结果] CSA刺激NRK52E细胞呈剂量依赖性抑制MMP-2活性和mRNA表达.与对照组(CSA 0 μmoL/L)比较,CSA0.42,0.84,4.2,8.4 μmoL/L的MMP-2活性分别下降为27%,24%,11%,9%:差异均有统计学意义,P<0.05.CSA刺激NRK52E细胞增加MMP-9活性和mRNA表达.与对照组比较,CSA 4.2 μmoL/L刺激48 h MMP-9活性上调为438%,72 h上调为237%,差异均有统计学意义,P<0.05.[结论] CSA对肾小管上皮细胞MMP-2表达和活性呈剂量依赖性抑制,以及对MMP-9表达和活性的诱导作用可能与CSA所致肾小管间质损害相关.
[목적]관찰배포소A(CSA)대근단신소관상피세포기질금속단백매-2화-9(MMP-2화MMP-9)표체화활성적영향.[방법]체외배양대서근단신소관상피세포주NRK52E세포지완전융합시,분별급여불동농도적CSA 0,0.42,0.84,4.2,8.4 μmoL/L,실험관찰48 h화72 h.수집배양액,용명효매보법검측MMP-2화MMP-9활성.제취총RNA,역전록취합매련식반응(RT-PCR)검측MMP-2화MMP-9적mRNA수평.[결과] CSA자격NRK52E세포정제량의뢰성억제MMP-2활성화mRNA표체.여대조조(CSA 0 μmoL/L)비교,CSA0.42,0.84,4.2,8.4 μmoL/L적MMP-2활성분별하강위27%,24%,11%,9%:차이균유통계학의의,P<0.05.CSA자격NRK52E세포증가MMP-9활성화mRNA표체.여대조조비교,CSA 4.2 μmoL/L자격48 h MMP-9활성상조위438%,72 h상조위237%,차이균유통계학의의,P<0.05.[결론] CSA대신소관상피세포MMP-2표체화활성정제량의뢰성억제,이급대MMP-9표체화활성적유도작용가능여CSA소치신소관간질손해상관.
[Objective] To observe the effect of cyclosporine A (CSA) on the activity and expression of MMP-2 and MMP-9 in renal tubular epithelial cells. [Methods] NRK52E cells were cultured until its reached confluent. Then NRK52E cells were exposed to different concentration of CSA (0, 0.42, 0.84, 4.2, and 8.4 μmoL/L) for 48 h or 72 h respectively. MMP-2 and MMP-9 activities were detected by gelatin zymography. The expression of MMP-2 and MMP-9 mRNA were detected by RT-PCR. [Results] MMP-2 activity and mRNA levels were decreased in a dose dependent manner after exposed to different concentration of CSA for 48 h or 72 h in NRK52E cells. Compared with control (CSA 0 μmoL/L), CSA 0.42, 0.84, 4.2, 8.4 μmoL/L decreased the MMP-2 activities to 27%, 24%, 11%, and 9% respectively; The differences are significant, P<0.05. But the MMP-9 activity and mRNA levels were increased after exposed to CSA for 48 h or 72 h in NRK52E cells. Compared with control group, CSA 4.2 μmoL/L exposure increased MMP-9 activity to 438% in 48 h, and 237% in 72 h; the differences are significant as well, P<0.05. [Conclusion] A dose-dependent decrease in the expression and activity of MMP-2, and the up-regulation of the expression and activity of MMP-9 by CSA in renal epithelial cells may related to CSA associated tubulointerstitial damage.