医用臭氧对大鼠体外星形胶质细胞的毒性

目的 评价医用臭氧对大鼠体外星形胶质细胞的毒性.方法 Wistar大鼠12只,日龄1～2 d,麻醉下取脑,分离、培养星形胶质细胞,接种入24孔培养板,每孔1 ml,细胞浓度7 × 105/ml,分为4组(n=7):正常对照组(C组)加入无任何干预的完全培养基400 μl;不同浓度医用臭氧组(O2-O340组、O2-O360组、O2-O380组)分别加入经40、60、80 μg/ml医用臭氧干预的完全培养基400 μl,于孵育2 h(T1)、4 h(T2)时观察细胞形态,测定星形胶质细胞超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量及乳酸脱氢酶(LDH)漏出率.结果 O2-O360组与O2-O380组胞体肥大、肿胀、突起增多,胞浆内出现黑色变性颗粒及空泡样改变,核固缩细胞增多,O2-O380组更明显.与C组比较,T1时O2-O340组SOD活性和MDA含量升高,LDH漏出率降低,O2-O360组SOD活性和MDA含量升高,O2-O380组SOD活性和MDA含量升高,LDH漏出率升高,T2时O2-O340组SOD活性升高,MDA含量和LDH漏出率降低,O2-O360组SOD活性和LDH漏出率升高,O2-O380组MDA含量和LDH漏出率升高(P<0.01);O2-O340组、O2-O360组和O2-O380组T1时MDA含量和LDH漏出率依次升高,T2时SOD活性依次降低,MDA含量和LDH漏出率依次升高(P<0.01);与T1时比较,T2时O2-O340组、O2-O360组和O2-O380组SOD活性和MDA含量降低,O2-O360组和O2-O380组LDH漏出率升高(P<0.05).结论 医用臭氧对大鼠体外星形胶质细胞的毒性作用与其浓度和作用时间有关.

Toxic effects of ozone on rat astrocytes in vitro

Objective To investigate the toxic effects of ozone on rat astrocytes in vitro. Methods Twelve 1-2 day old Wistar rats were anesthetized with ether. Their brains were removed. Astrecytes were enzymatically isolated with trypsin and cultured. After different-speed cultivation and serial passage, astrocytes were seeded into 24 well plates and divided into 4 groups (n = 7 each): group Ⅰ normal control (group C) and group Ⅱ , Ⅲ , Ⅳ (group O2-O340, 60, 80) were exposed to ozone 40, 60 and 80 μg/ml respectively. After 2 and 4 h incubation, the astroeytes were examined under microscope. Supernatant was collected and cells were broken for determination of SOD activity and MDA content in the cells and LDH leaking ratio. Results Cells in group O2-O3 60 and 80 exhibited hypertrophy, vacuoles and black denatured grains in the cytoplasm and increase in proceses. SOD activity and MDA content were significantly increased at the end of 2 h incubation in group O2-O340, 60 and 80 as compared with blank control group. LDH leaking ratio and MDA content were increased in a concentration dependent manner at the end of 2 h and 4 h incubation, while SOD activity was decreased concentration dependently at the end of 4 h incubation. SOD activity was signifieandy decreased, while LDH leaking ratio was increased at 4 h as compared with that at 2 h in group O2-O360 and 80. Conclusion Ozone has toxic effects on cultured rat astroeytns in a concentration and time dependent manner.