Protective Effect of Protease Inhibitor OH-TCI Against Myocardial Ischemia Reperfusion Injury in Rats
- Author:
Yuan-Yuan GAO
1
Author Information
- Publication Type:Journal Article
- Keywords: Myocardial ischemia; Myocardial protection; OH-TCI; Reperfusion injury
- From: Chinese Pharmaceutical Journal 2017;52(20):1837-1841
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To investigate the expression and purification of protease inhibitor OH-TCI in Escherichia coli and its protective effect and mechanism in myocardial ischemia-reperfusion injury model. METHODS: The recombinant plasmid of Pet32a-OH-TCI was transformed into E. coli and induced by IPTG, soluble fusion protein was expressed in the constructed expression system and isolated by a nickel affinity chromatography column. Then, the products was digested by TEV enzyme and was purified by the nickel affinity chromatography column. The adult male rats were divided into two groups randomly: normal saline group and OH-TCI group. At 5 min before the operation, rats were treated with OH-TCI by intraperitoneal injection in the experimental group and was given the same volume of saline in the control group. In all animal experiments, rats left anterior descending coronary artery were ligated for 30 min to construct the model. After 120 min of reperfusion, samples were taken to determine the contents of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukins-6 (IL-6) and the activities of lactic acid dehydrogenase (LDH), creatine kinase (CK) and superoxide dismutase (SOD). Finally, the protective effects of OH-TCI against myocardial ischemia-reperfusion injury in rat model were evaluated. RESULTS: The recombinant protease inhibitor OH-TCI was expressed in the E.coli expression system in a soluble pattern and successfully purified by two steps of nickel affinity chromatography. Comparing with those in the control group, the recombinant protease inhibitor OH-TCI could significantly decrease the levels of LDH, MDA and IL-6 (P<0.01)as well as inflammatory factor TNF-α (P<0.05). CONCLUSION: The recombinant protease inhibitor OH-TCI can be expressed successfully in E. coli. Purified recombinant OH-TCI has natural protease inhibitory activity and shows protective effects against ischemia and reperfusion myocardium injury.
