外源性内皮素-1对原代培养新生大鼠心肌细胞的影响

EFFECTS OF EXOGENOUS ENDOT HELIN-1 ON CULTURED PRIMARY CARDIOMYOCYTES FROM NEONATAL RATS

①目的 以原代培养的新生大鼠心肌细胞为模型 ,探讨外源性内皮素 1对心肌细胞的损伤作用。②方法 将原代培养的心肌细胞分成 4组 ,实验组分别使原代培养的心肌细胞培养液内皮素 1的浓度达 10 -7、10 -8、10 -9mol/L(A、B、C组 ) ,正常对照组换不含血清的培养液培养 ,分别于实验前和实验 12、2 4h ,取上清液 - 2 0℃冻存待测。用倒置相差显微镜对活细胞形态进行观察 ,用Hitachi 715 0全自动生化仪测定培养液中心肌细胞乳酸脱氢酶 (LDH)的活性 ,用微量邻苯三酚法测定心肌细胞超氧化物歧化酶 (SOD)的活性 ,用丙二醛 (MDA)测试盒测定培养液中MDA的含量。③结果 实验组较对照组心肌细胞形态及搏动特点有异常改变 ,内皮素 1(10 -7、10 -8、10 -9mol/L)作用 12或 2 4h后 ,同簇心肌细胞搏动节律变得不规整 ,心肌细胞胞体立体感差 ,胞浆也显得粗糙 ,成纤维细胞数目增多。实验组较对照组培养液中心肌细胞LDH的活性明显增加 ,差异有显著性 (F =5 5 .2 5、73.4 6 ,q =3.6 5～ 19.18,P <0 .0 5 )。实验组较对照组培养液中心肌细胞SOD的活性明显下降 ,差异有显著性 (F =5 2 .4 3、75 .82 ,q =4 .2 3～ 19.4 7,P <0 .0 5 )。实验组较对照组培养液中MDA的含量明显增加 ,差异有显著性 (F =15 2 .6 2、5 4 8.5 4 。

Objective\ To study the e ffects of exogenous endothelin 1 on the cultured primary cardiomyocytes from ne onatal rats.\ Methods\ The cultured primary cardiomyocytes were randomly divided into endothelin 1 (ET 1) group and the control group. The ET 1 group was sub divided into 3 subgroups, with the concentration of exogenous endothelin 1 in t he cardiomyocyte culture being 10 -9 ,10 -8 ,and 10 -7 mol/L, res pectively. The cells in the control group were cultured without serum. The super nates were taken and frozen at -20 ℃ for detection before the experiment and 12 and 24 hours after the experiment. Morphological changes were observed with a microtelescope, and the levels of LDH?MDA and SOD in the supernate were examine d with physiochemical techniques.\ Results\ Compared with those in the control group, the morphologic al and beating characters of cardiomyocytes in the ET 1 groups underwent abnorm al changes; the levels of LDH in the supernate increased significantly in the ET 1 subjects ( F=55.25, 73.46; q=3.65-19.18; P <0.05); the levels of SOD dec reased significantly ( F=52.43,75.82; q=4.23- 19.47 ; P <0.05); and the le vels of MDA increased significantly ( F=152.62,548.54; q=3.94-50.84; P <0.05 ).\ Conclusion Exogenous endothelin 1 can injure cultured primary cardiomyocytes from neonatal rats, causing the abnormality in cellular structur es and metabolism.