CAJ | 학술논문

目的探讨红景天苷(Sa)对大鼠心肌缺血/再灌注(I/R)损伤后内皮素-1(ET-1)的影响及其保护机制.方法将60只 Wistar 大鼠按随机数字表法均分为假手术组、缺血组、I/R 1 h 组、I/R 2 h 组、Sa 预处理1 h 组和 Sa 预处理2 h 组.采用结扎冠状动脉(冠脉)前降支30 min 后恢复血流灌注复制 I/R 损伤大鼠模型.Sa 组于术前20 min 舌下静脉给予 Sa 25 mg/kg 预处理.相应时间点观察假手术组、缺血组、I/R 组心电图 ST 段抬高幅度;采用酶联免疫吸附试验(ELISA)测定各组血清肌酸激酶(CK)、乳酸脱氢酶(LDH)的活性,放射免疫试验测定血清内皮素-1(ET-1)水平,逆转录-聚合酶链反应(RT-PCR)、蛋白质免疫印迹法(Western blotting)检测各组心肌组织 ET-1 mRNA 及蛋白表达.结果缺血组和 I/R 组 ST 段抬高幅度及血清 CK、LDH 均较假手术组显著升高;I/R 组 ST 段抬高幅度较缺血组显著降低,CK、LDH 则较缺血组显著升高.与假手术组比较,缺血组、I/R 1 h 组、I/R 2 h 组、Sa 预处理1 h 组、Sa 预处理2 h 组血清 ET-1(μg/L)水平均明显升高(66.49±8.61、104.60±6.64、108.82±6.22、71.30±12.22、69.60±7.40比24.58±3.41,均P<0.05);心肌 ET-1蛋白表达(灰度值)明显升高(0.51±0.07、0.76±0.08、0.77±0.06、0.53±0.09、0.58±0.04比0.34±0.10,均 P<0.05);I/R 1 h 组、I/R 2 h 组、Sa 预处理1 h 组、Sa 预处理2 h 组 ET-1 mRNA 表达〔吸光度(A)值〕均明显升高(0.73±0.04、0.78±0.02、0.51±0.06、0.55±0.03比0.31±0.07,均 P<0.05),而缺血组则差异无统计学意义(P>0.05).与缺血组比较,I/R 1 h 组、I/R 2 h 组血清 ET-1水平、心肌组织 ET-1 mRNA 和蛋白表达均明显升高(均 P<0.05);Sa 预处理1 h组、Sa 预处理2 h 组血清 ET 水平及心肌组织 ET-1 mRNA 和蛋白表达均较 I/R 1 h 组、I/R 2 h 组明显下降(均P<0.05).结论 ET-1在大鼠 I/R 损伤过程中表达上调,用 Sa 预处理可显著降低 ET-1的表达,间接起到保护心肌的作用. 目的探討紅景天苷(Sa)對大鼠心肌缺血/再灌註(I/R)損傷後內皮素-1(ET-1)的影響及其保護機製.方法將60隻 Wistar 大鼠按隨機數字錶法均分為假手術組、缺血組、I/R 1 h 組、I/R 2 h 組、Sa 預處理1 h 組和 Sa 預處理2 h 組.採用結扎冠狀動脈(冠脈)前降支30 min 後恢複血流灌註複製 I/R 損傷大鼠模型.Sa 組于術前20 min 舌下靜脈給予 Sa 25 mg/kg 預處理.相應時間點觀察假手術組、缺血組、I/R 組心電圖 ST 段抬高幅度;採用酶聯免疫吸附試驗(ELISA)測定各組血清肌痠激酶(CK)、乳痠脫氫酶(LDH)的活性,放射免疫試驗測定血清內皮素-1(ET-1)水平,逆轉錄-聚閤酶鏈反應(RT-PCR)、蛋白質免疫印跡法(Western blotting)檢測各組心肌組織 ET-1 mRNA 及蛋白錶達.結果缺血組和 I/R 組 ST 段抬高幅度及血清 CK、LDH 均較假手術組顯著升高;I/R 組 ST 段抬高幅度較缺血組顯著降低,CK、LDH 則較缺血組顯著升高.與假手術組比較,缺血組、I/R 1 h 組、I/R 2 h 組、Sa 預處理1 h 組、Sa 預處理2 h 組血清 ET-1(μg/L)水平均明顯升高(66.49±8.61、104.60±6.64、108.82±6.22、71.30±12.22、69.60±7.40比24.58±3.41,均P<0.05);心肌 ET-1蛋白錶達(灰度值)明顯升高(0.51±0.07、0.76±0.08、0.77±0.06、0.53±0.09、0.58±0.04比0.34±0.10,均 P<0.05);I/R 1 h 組、I/R 2 h 組、Sa 預處理1 h 組、Sa 預處理2 h 組 ET-1 mRNA 錶達〔吸光度(A)值〕均明顯升高(0.73±0.04、0.78±0.02、0.51±0.06、0.55±0.03比0.31±0.07,均 P<0.05),而缺血組則差異無統計學意義(P>0.05).與缺血組比較,I/R 1 h 組、I/R 2 h 組血清 ET-1水平、心肌組織 ET-1 mRNA 和蛋白錶達均明顯升高(均 P<0.05);Sa 預處理1 h組、Sa 預處理2 h 組血清 ET 水平及心肌組織 ET-1 mRNA 和蛋白錶達均較 I/R 1 h 組、I/R 2 h 組明顯下降(均P<0.05).結論 ET-1在大鼠 I/R 損傷過程中錶達上調,用 Sa 預處理可顯著降低 ET-1的錶達,間接起到保護心肌的作用.
목적탐토홍경천감(Sa)대대서심기결혈/재관주(I/R)손상후내피소-1(ET-1)적영향급기보호궤제.방법장60지 Wistar 대서안수궤수자표법균분위가수술조、결혈조、I/R 1 h 조、I/R 2 h 조、Sa 예처리1 h 조화 Sa 예처리2 h 조.채용결찰관상동맥(관맥)전강지30 min 후회복혈류관주복제 I/R 손상대서모형.Sa 조우술전20 min 설하정맥급여 Sa 25 mg/kg 예처리.상응시간점관찰가수술조、결혈조、I/R 조심전도 ST 단태고폭도;채용매련면역흡부시험(ELISA)측정각조혈청기산격매(CK)、유산탈경매(LDH)적활성,방사면역시험측정혈청내피소-1(ET-1)수평,역전록-취합매련반응(RT-PCR)、단백질면역인적법(Western blotting)검측각조심기조직 ET-1 mRNA 급단백표체.결과결혈조화 I/R 조 ST 단태고폭도급혈청 CK、LDH 균교가수술조현저승고;I/R 조 ST 단태고폭도교결혈조현저강저,CK、LDH 칙교결혈조현저승고.여가수술조비교,결혈조、I/R 1 h 조、I/R 2 h 조、Sa 예처리1 h 조、Sa 예처리2 h 조혈청 ET-1(μg/L)수평균명현승고(66.49±8.61、104.60±6.64、108.82±6.22、71.30±12.22、69.60±7.40비24.58±3.41,균P<0.05);심기 ET-1단백표체(회도치)명현승고(0.51±0.07、0.76±0.08、0.77±0.06、0.53±0.09、0.58±0.04비0.34±0.10,균 P<0.05);I/R 1 h 조、I/R 2 h 조、Sa 예처리1 h 조、Sa 예처리2 h 조 ET-1 mRNA 표체〔흡광도(A)치〕균명현승고(0.73±0.04、0.78±0.02、0.51±0.06、0.55±0.03비0.31±0.07,균 P<0.05),이결혈조칙차이무통계학의의(P>0.05).여결혈조비교,I/R 1 h 조、I/R 2 h 조혈청 ET-1수평、심기조직 ET-1 mRNA 화단백표체균명현승고(균 P<0.05);Sa 예처리1 h조、Sa 예처리2 h 조혈청 ET 수평급심기조직 ET-1 mRNA 화단백표체균교 I/R 1 h 조、I/R 2 h 조명현하강(균P<0.05).결론 ET-1재대서 I/R 손상과정중표체상조,용 Sa 예처리가현저강저 ET-1적표체,간접기도보호심기적작용.
Objective To investigate the effect of Salidroside(Sa)on changes of endothelin-1(ET-1)in rats after myocardial ischemia/reperfusion(I/R)injury and its protective mechanism. Methods Sixty Wistar rats were grouped randomly into sham group(S),ischemic 30 minutes group(I),ischemic 30 minutes + reperfusion 1-hour group(I/R 1 h),ischemic 30 minutes + reperfusion 2-hour group(I/R 2 h),Sa + ischemic 30 minutes + reperfusion 1-hour group(Sa+I/R 1 h)and Sa + ischemic 30 minutes + reperfusion 2-hour group(Sa+I/R 2 h). Myocardial I/R injury model was reproduced by 30-minute ligation of the left anterior descending branch,followed by reperfusion after ischemia. In Sa + I/R groups,25 mg/kg Sa was given in sublingual vein 20 minutes before surgery. The ST-segments amplitude of electrocardiogram(ECG)in S,I and various I/R groups was observed at different time points. The activities of creatine kinase(CK)and lactate dehydrogenase(LDH)were examined by enzyme-linked immunosorbent assay(ELISA). The changes of ET-1 were evaluated by radioimmunoassay,cardiac tissue ET-1 mRNA was determined by reverse transcriptase-polymerase chain reaction(RT-PCR)and its protein was measured by Western blotting. Results The ST-segments amplitude of ECG,CK and LDH in serum in I and I/R groups were increased significantly compared with those in S group. The magnitude of ST-segment elevation was lower significantly in I/R groups than that in I group,while CK and LDH in serum in I/R groups were obviously higher than those in I group. Compared with S group,the serum levels of ET-1(μg/L)were increased in groups of I,I/R 1 h,I/R 2 h,Sa + I/R 1 h and Sa + I/R 2 h(66.49±8.61,104.60±6.64,108.82±6.22,71.30±12.22,69.60±7.40 vs. 24.58±3.41,allP<0.05), the protein expression of ET-1(gray scale)in myocardial tissue was increased significantly(0.51±0.07,0.76±0.08, 0.77±0.06,0.53±0.09,0.58±0.04 vs. 0.34±0.10,all P<0.05),the expression of ET-1 mRNA 〔absorbance(A) value〕 was increased significantly in groups of I/R 1 h,I/R 2 h,Sa + I/R 1 h and Sa + I/R 2 h(0.73±0.04,0.78±0.02, 0.51±0.06,0.55±0.03 vs. 0.31±0.07,all P<0.05),and the difference in ET-1 mRNA between I group and S group was not statistically significant(P>0.05). Compared with I group,the serum ET-1 level,and the expression levels of cardiac tissue ET-1 mRNA and protein were increased significantly in groups of I/R 1 h and I/R 2 h(all P<0.05), and the serum ET-1 level and the expression levels of cardiac tissue ET-1mRNA and protein in groups of Sa + I/R 1 h and Sa + I/R 2 h were lower significantly than those in I/R 1 h and I/R 2 h groups(all P<0.05). Conclusions ET-1 is up-regulated in myocardial ischemia and reperfusion injury in rats. The pre-treatment of Sa can significantly reduce the expression of ET-1 and indirectly play a role of protecting the myocardial tissue.