PARP及NF-κB在重症急性胰腺炎大鼠肾上腺细胞中的表达及5-AIQ/PDTC的干预作用
Expression of PARP/NF-κB and intervention effect of 5-AIQ/PDTC in SAP rats with adrenal damage
目的 探讨ADP核糖聚合酶(PARP)/核因子(NF)-κB在大鼠重症急性胰腺炎(SAP)肾上腺损伤中的表达情况及PARP抑制剂5-氨基异喹啉酮(5-AIQ)/吡咯烷二硫代氨基甲酸盐(PDTC)的干预作用.方法 将原代培养的肾上腺皮质细胞进行细胞定量后分为6组:空白对照(SO组)、胰腺炎组(SAP组)、PDTC药物对照组(SO+ PDTC组)、PDTC干预组(SAP+ PDTC组)、5-AIQ药物对照组(SO +5-AIQ组)和5-AIQ干预组(SAP+ 5-AIQ组).以胰腺炎大鼠血清刺激SAP组和2个干预组,之后以最适浓度加入每组相应药物,在给药后继续培养12h,观察各组细胞皮质酮分泌情况及PARP/NF-κB的表达情况.结果 体外培养肾上腺细胞呈圆形或类圆形,胞质内有分泌颗粒,培养第2天贴壁率约60%,可被3β-HSD染色.SAP组皮质酮水平为(216.4±15.7) μg/L,明显低于SO组(294.8±16.3)μg/L(P <0.05),5-AIQ干预组和PDTC干预组皮质酮水平分别为(258.6±19.0)μg/L及(264.3±18.2)μg/L,明显高于SAP组但低于SO组和药物对照组(P <0.05);SAP组和PDTC干预组肾上腺细胞PARP-1的表达高于SO组,5-AIQ干预组PARP-1的表达明显低于SAP组和PDTC干预组,但高于SO组和药物对照组;SAP组肾上腺细胞NF-κB的表达高于SO组,5-AIQ干预组和PDTC干预组NF-κB的表达明显低于SAP组,但高于SO组和药物对照组.结论 PARP/NK-κB通路参与重症急性胰腺炎肾上腺损伤过程,应用5-AIQ及PDTC可在一定程度上减轻肾上腺细胞功能的损伤程度.
更多Objective To explore the expression of poly (ADP-ribose) polymerase/nuclear factor-κB (PARP/NF-κB) and intervention effect of 5-aminoisoquinolinone/pyrrolidine dithiocarbamate (5-AIQ/PDTC) in severe acute pancreatitis (SAP) rats with adrenal damage.Methods The primarily cultured adrenocortical cells were quantitatively divided into control group (SO),pancreatitis group (SAP),PDTC drug control group (SO + PDTC),PDTC intervention group (SAP + PDTC),5-AIQ drug control group (SO +5-AIQ) and 5-AIQ intervention group (SAP + 5-AIQ).The SAP and 2 intervention groups were stimulated with the sera of SAP rats.Then corresponding drugs were added and culture continued for 12 hours.The corticosterone levels and PARP/NF-κB expression were observed for each group.Results Adrenal cells in vitro cultured were round or oval,had secretory granules and could be stained by 3β-hydroxysteroid dehydrogenase antibody.The adherence rate was 60% after 48-hour culturing.The corticosterone level of SAP group was significantly lower than that of SO group [(216.4 ± 15.7) vs (294.8 ± 16.3) μg/L,P < 0.05].The 2 intervention groups were (258.6 ± 19.0) and (264.3 ± 18.2) μg/L respectively.These two values were higher than those of SAP group (P < 0.05),but lower than those of SO group (P < 0.05).With regards to the expression of PARP-1,the SAP and PDTC intervention groups were higher than SO group while 5-AIQ intervention group was significantly lower than SAP and PDTC intervention groups,but higher than SO and drug control groups.The expression of NF-κB in SAP group was higher than that in SO group.Two intervention groups were lower than SAP group,but higher than SO and drug control groups.Conclusion The pathway of PARP/NF-κB participates in adrenal damage of SAP rats.To a certain extent,the uses of 5-AIQ and PDTC may alleviate adrenal damage.
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