CAJ | 학술논문

目的： Rho 激酶介导的肌动蛋白细胞骨架重组在肌成纤维细胞(MFB)表型分化中具有重要作用。本实验拟观察 Rho 激酶抑制剂法舒地尔对压力超负荷大鼠心脏 MFB 表型分化的影响。方法采用腹主动脉缩窄法建立压力超负荷诱导的心肌纤维化大鼠模型。假手术组(Sham 组)仅分离但不予结扎腹主动脉。术后4周末,将存活大鼠按随机数表法分为模型组(Model 组)、法舒地尔高剂量组(FH 组)和法舒地尔低剂量组(FL 组)。药物干预4周末,观察大鼠心肌病理学改变,并检测心肌羟脯氨酸(HYP)含量,评估心肌纤维化程度；酶联免疫吸附法检测血浆和心肌血管紧张素Ⅱ(AngⅡ)含量；免疫组织化学法分析心肌磷酸化肌球蛋白磷酸酶靶蛋白亚基1(p-MYPT1)、α-平滑肌肌动蛋白(α-SMA)、骨桥蛋白(OPN)和转化生长因子β1(TGF-β1)的表达。结果 Model 组大鼠心肌纤维化程度较 Sham 组明显加重,AngⅡ含量显著升高,p-MYPT1(0.063±0.009比0.029±0.014)、α-SMA(0.034±0.009比0.004±0.003)、OPN(0.043±0.007比0.012±0.006)和 TGF-β1(0.058±0.019比0.019±0.009)的表达均明显升高(均为 P ﹤0.01)； FH 组和 FL 组大鼠心肌纤维化程度较Model 组减轻,AngⅡ含量均显著下降,p-MYPT1(0.029±0.013和0.040±0.011)、α-SMA(0.016±0.006和0.027±0.007)、OPN(0.018±0.004和0.036±0.006)和 TGF-β1(0.022±0.013和0.039±0.014)的表达也不同程度下降(P ﹤0.01或 P ﹤0.05)。结论法舒地尔改善压力超负荷大鼠心肌纤维化的作用至少与部分抑制 MFB 表型分化有关。
목적： Rho 격매개도적기동단백세포골가중조재기성섬유세포(MFB)표형분화중구유중요작용。본실험의관찰 Rho 격매억제제법서지이대압력초부하대서심장 MFB 표형분화적영향。방법채용복주동맥축착법건립압력초부하유도적심기섬유화대서모형。가수술조(Sham 조)부분리단불여결찰복주동맥。술후4주말,장존활대서안수궤수표법분위모형조(Model 조)、법서지이고제량조(FH 조)화법서지이저제량조(FL 조)。약물간예4주말,관찰대서심기병이학개변,병검측심기간포안산(HYP)함량,평고심기섬유화정도；매련면역흡부법검측혈장화심기혈관긴장소Ⅱ(AngⅡ)함량；면역조직화학법분석심기린산화기구단백린산매파단백아기1(p-MYPT1)、α-평활기기동단백(α-SMA)、골교단백(OPN)화전화생장인자β1(TGF-β1)적표체。결과 Model 조대서심기섬유화정도교 Sham 조명현가중,AngⅡ함량현저승고,p-MYPT1(0.063±0.009비0.029±0.014)、α-SMA(0.034±0.009비0.004±0.003)、OPN(0.043±0.007비0.012±0.006)화 TGF-β1(0.058±0.019비0.019±0.009)적표체균명현승고(균위 P ﹤0.01)； FH 조화 FL 조대서심기섬유화정도교Model 조감경,AngⅡ함량균현저하강,p-MYPT1(0.029±0.013화0.040±0.011)、α-SMA(0.016±0.006화0.027±0.007)、OPN(0.018±0.004화0.036±0.006)화 TGF-β1(0.022±0.013화0.039±0.014)적표체야불동정도하강(P ﹤0.01혹 P ﹤0.05)。결론법서지이개선압력초부하대서심기섬유화적작용지소여부분억제 MFB 표형분화유관。
Objective Rho kinase mediated cytoskeleton actin reorganization plays an important role in the phenotypic differentiation of myofibroblasts. The study is to investigate the effects of Fasudil---a Rho kinase inhibitor---on myofibroblasts differentiation in pressure overload rats. Methods Pressure overload induced myocardial fibrosis rats models were established by abdominal aortic constriction. In sham group the abdominal aorta was just separated but not ligated. The 4 weeks after surgery, the operation survival rats were randomly divided into 3 groups: control model group, Fasudil high dose group (FH group) and Fasudil low dose group (FL group). And 4 weeks after the drug intervention, the degree of myocardial fibrosis was assessed as follows: pathologic changes were observed and hydroxyproline contents of the myocardium were determined; the content of angiotensin Ⅱ ( Ang-Ⅱ) was calculated by enzyme linked immunosorbent assay (Elisa); the expression levels of phosphorylated myosin phosphatase target subunit 1 (p-MYPT1), α-smooth muscle actin ( α-SMA), osteopontin ( OPN) and transforming growth factor-β1 (TGF-β1) were analyzed by immunohistochemistry. Results Compared with the sham group, the degree of myocardial fibrosis in model group was aggravated variously, and the content of Ang-Ⅱ and the expression levels of p-MYPT1 (0. 063 ± 0. 009 vs. 0. 029 ± 0. 014), α-SMA (0. 034 ± 0. 009 vs. 0. 004 ± 0. 003), OPN (0. 043 ± 0. 007 vs. 0. 012 ± 0. 006 ) and TGF-β1 ( 0. 058 ± 0. 019 vs. 0. 019 ± 0. 009 ) were significantly increased (all P ﹤ 0. 01). Compared with the model group, the degree of myocardial fibrosis in FH group and FL group were relieved, and the content of Ang-Ⅱ and the expression levels of p-MYPT1 (0. 029 ± 0. 013, 0. 040 ± 0. 011), α-SMA (0. 016 ± 0. 006, 0. 027 ± 0. 007), OPN (0. 018 ± 0. 004, 0. 036 ± 0. 006) and TGF-β1 (0. 022 ± 0. 013, 0. 039 ± 0. 014) were significantly decreased (P ﹤ 0. 05 or P ﹤ 0. 01). Conclusions The protection effect of Fasudil on myocardial fibrosis in pressure overloaded rats is at least partially related to the inhibition of myofibroblast differentiation.