乌司他丁对缺氧诱导PC12细胞神经损伤的保护作用研究  

Protective effect of ulinastatin against hypoxia-induced injury in PC12 nerve cells

作　　者：王德佳[1] 杨波[1] WANG De-Gui;YANG Bo(Chengde Central Hospital,Chengde,Hebei 067000,China)

机构地区：[1]承德市中心医院,河北承德067000

出　　处：《国际神经病学神经外科学杂志》2020年第5期469-474,共6页Journal of International Neurology and Neurosurgery

摘　　要：目的探讨乌司他丁对缺氧诱导PC12细胞神经损伤的作用及分子机制。方法PC12细胞分为对照组、缺氧组、缺氧+乌司他丁低、中、高剂量组、缺氧+miR-NC组、缺氧+miR-190组、缺氧+乌司他丁+anti-miR-NC组、缺氧+乌司他丁+anti-miR-190组。检测培养液中乳酸脱氢酶(LDH)漏出率及细胞中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;流式细胞术检测细胞凋亡;蛋白质印迹法(Western blotting)检测蛋白表达;实时荧光定量聚合酶链反应(qRT-PCR)检测miR-190表达水平。结果与缺氧组比较,缺氧+乌司他丁低、中、高剂量组PC12细胞中LDH漏出率降低,SOD活性升高,MDA含量降低,细胞凋亡率降低,BcI-2相对表达量升高,Bax相对表达量降低,miR-190相对表达量升高,且呈剂量依赖性(均P<0.05)。miR-190过表达后,LDH漏出率降低,SOD活性升高,MDA含量降低,细胞凋亡率降低,BcI-2相对表达量升高,Bax相对表达量降低(均P<0.05)。抑制miR-190表达能逆转乌司他丁对缺氧诱导的PC12细胞损伤的作用。结论乌司他丁可能通过上调miR-190表达对缺氧诱导PC12细胞神经损伤起保护作用。Objective To investigate the effect of ulinastatin on hypoxia-induced injury in PC12 nerve cells and its molecular mechanism.Methods PC12 cells were divided into control group,hypoxia group,hypoxia+low/middle/high-dose ulinastatin groups,hypoxia+miR-NC group,hypoxia+miR-190 group,hypoxia+ulinastatin+anti-miR-NC group,and hypoxia+ulinastatin+anti-miR-190 group.Related kits were used to measure the leakage rate of lactate dehydrogenase(LDH)in the culture medium and the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in cells;flow cytometry was used to measure cell apoptosis;Western blotting was used to measure protein expression;quantitative real-time PCR was used to measure the expression of miR-190.Results Compared with the hypoxia group,the hypoxia+low/middle/high-dose ulinastatin groups had significant reductions in LDH leakage rate,MDA content,and cell apoptosis rate and a significant increase in SOD activity,as well as significant increases in the relative expression of Bcl-2 and miR-190 and a significant reduction in the relative expression of Bax,in a dose-dependent manner(all P<0.05).After miR-190 overexpression,there was a significant reduction in LDH leakage rate,a significant increase in SOD activity,significant reductions in MDA content and cell apoptosis rate,a significant increase in the relative expression of Bcl-2,and a signify icant reduction in the relative expression of Bax(all P<0.05).Inhibition of the expression of miR-190 reversed the effect o£ulinastatin on hypoxia-induced injury of PC12 cells.Conclusions Ulinastatin can exert a protective effect against hypoxia-induced injury of PC12 nerve cells possibly by upregulating the expression of miR-190.

关 键 词：乌司他丁 miR-190 神经细胞损伤 氧化应激 凋亡 

分 类 号：R741[医药卫生—神经病学与精神病学]