CAJ | 학술논문

目的探讨茯苓酸(PA)改善梗阻性肾积水后导致的肾间质纤维化的作用及其机制.方法将28只C57雄性小鼠按照随机列表法分为4组:A组:假手术组;B组:模型组;C组:茯苓酸小剂量组;D组:茯苓酸大剂量组.建模及给药7d后,处死小鼠并取材,苏木素-伊红(HE)染色观察肾脏病理损伤变化,天狼猩红染色检测间质胶原纤维的表达,免疫组织化学染色及实时定量聚合酶链反应(Real-time PCR)检测间质中胶原纤维蛋白-1(COI-1)、纤维粘连蛋白(FN)和平滑肌肌动蛋白(α-SMA)的表达,Western blot检测转化生长因子-β1(TGF-β1),磷酸化Smad2/3(pSmad2/3),基质金属蛋白酶-9(MMP-9)的表达.结果输尿管梗阻(UUO)小鼠给予茯苓酸治疗7d后,HE染色显示肾脏病理改变较对照组减轻.天狼猩红染色可见肾间质纤维化程度较对照组明显减轻,且大剂量组减轻更明显.免疫组织化学结果显示COI-1、FN、α-SMA在梗阻后表达明显升高,给予茯苓酸治疗可降低基质中COI-1、FN、α-SMA的表达.Real-time PCR结果显示,在模型组、小剂量茯苓酸组、大剂量茯苓酸组COI-1 mRNA水平相对表达量分别为4.20±1.42、2.30±0.76、1.70 ±0.70;FN在mRNA水平相对表达量分别为5.20 ±2.22、3.00±1.70、1.70±1.06;α-SMA在mRNA水平相对表达量分别4.60±1.23、2.30 ±0.98、1.40 ±0.70.给药组与模型组比较,差异有统计学意义(P＜0.05).Western blot结果显示茯苓酸可下调TGF-β1、pSmad2/3、MMP-9的表达.结论茯苓酸可改善单侧肾积水导致的间质纤维化,可能是通过下调TGF-β1、pSmad2/3、MMP-9的表达来实现的.
목적 탐토복령산(PA)개선경조성신적수후도치적신간질섬유화적작용급기궤제.방법 장28지C57웅성소서안조수궤렬표법분위4조:A조:가수술조;B조:모형조;C조:복령산소제량조;D조:복령산대제량조.건모급급약7d후,처사소서병취재,소목소-이홍(HE)염색관찰신장병리손상변화,천랑성홍염색검측간질효원섬유적표체,면역조직화학염색급실시정량취합매련반응(Real-time PCR)검측간질중효원섬유단백-1(COI-1)、섬유점련단백(FN)화평활기기동단백(α-SMA)적표체,Western blot검측전화생장인자-β1(TGF-β1),린산화Smad2/3(pSmad2/3),기질금속단백매-9(MMP-9)적표체.결과 수뇨관경조(UUO)소서급여복령산치료7d후,HE염색현시신장병리개변교대조조감경.천랑성홍염색가견신간질섬유화정도교대조조명현감경,차대제량조감경경명현.면역조직화학결과현시COI-1、FN、α-SMA재경조후표체명현승고,급여복령산치료가강저기질중COI-1、FN、α-SMA적표체.Real-time PCR결과현시,재모형조、소제량복령산조、대제량복령산조COI-1 mRNA수평상대표체량분별위4.20±1.42、2.30±0.76、1.70 ±0.70;FN재mRNA수평상대표체량분별위5.20 ±2.22、3.00±1.70、1.70±1.06;α-SMA재mRNA수평상대표체량분별4.60±1.23、2.30 ±0.98、1.40 ±0.70.급약조여모형조비교,차이유통계학의의(P＜0.05).Western blot결과현시복령산가하조TGF-β1、pSmad2/3、MMP-9적표체.결론복령산가개선단측신적수도치적간질섬유화,가능시통과하조TGF-β1、pSmad2/3、MMP-9적표체래실현적.
Objective To investigate protective effects of pachymic acid (PA) on renal interstitial fibrosis induced by hydronephrosis of obstructive models and its potential mechanism.Methods Twentyeight male C57 mice were divided into four groups according to random list method,each containing seven mice.A:sham operation group; B:model group; C:a small dose pachymic acid group; D:a high dose pachymic acid group.All mice were sacrificed on 7 days after operation and tubular injury was observed using hematoxylin and eosin (HE) staining.Sirius red staining was performed to detect the expression of collagen in interstitial,and immunohistochemistry and real-time polymerase chain reaction (Real-time PCR) were used to assay mRNA expressions of collagen-1 (COI-1),fibronectin (FN) and α-smooth muscle actin (α-SMA).Transforming growth factor-β1 (TGF-β1),pSmad2/3,matrix metalloproteinase-9 (MMP-9) at protein levels were measured using Western blotting.Results Seven days after the treatment of pachymic acid in unilateral ureteral obstruction (UUO) model,HE staining indicated that renal tubular injury was ameliorated in the PA treated group,and the results of Sirius red staining showed that renal interstitial fibrosis were significantly reduced compared to model group,and it is more obvious in high dose group.The results of immunohistochemistry indicated that COI-1,FN,α-SMA expressions were significantly increased in model group,and the treatment of pachymic acid significantly inhibited the expressions of COI-1,FN and α-SMA in matrix.Meanwhile,mRNA levels of COI-1 detected by Real-time PCR in model group,the small dose of pachymic acid group and the large dose of pachymic acid group were 4.20 ± 1.42,2.30 ± 0.76,1.70 ± 0.70.Expressions of FN in three groups were 5.20 ± 2.22,3.00 ± 1.70,1.70 ± 1.06 and α-SMA expressions were 4.60 ± 1.23,2.30 ± 0.98,1.40 ± 0.70.There was statistical difference compared with the model group.The results of Western blotting showed pachymic acid could down-regulate expressions of TGF-β1,pSmad2/3 and MMP-9.Conclusion Pachymic acid could improve interstitial fibrosis induced by unilateral hydronephrosis,which might be achieved through the approach of suppressing the expressions of TGF-β1,pSmad2/3 and MMP-9.