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      Establishment of THP-1 Monocytes with Compromised Mitochondrial Functions

      Chou, Tzu-wen Joy
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      Chou, T. J. (2009). Establishment of THP-1 Monocytes with Compromised Mitochondrial Functions (Thesis, Master of Science (MSc)). The University of Waikato, Hamilton, New Zealand. Retrieved from https://hdl.handle.net/10289/4569
      Permanent Research Commons link: https://hdl.handle.net/10289/4569
      Abstract
      The process of inflammation is important for both normal health and in a number of diseases, such as metabolic disorders, autoimmune diseases, and neurodegenerative diseases. Mitochondria are vital for the functioning of all cells. It had been implicated as a key player in inflammatory processes, especially through reactive oxygen species as signals of various immune responses. This study aimed to establish a THP-1 cell line with compromised mitochondrial functions, using antimycin A as a Complex III inhibitor, and to investigate the role of mitochondrial stress, as monitored by the expression of Hsp60, in inflammatory processes. High concentrations of antimycin A (100 and 200 μM) were cytotoxic to THP-1 monocytes that they were rapidly killed within 48 hours of exposure. Lower concentrations of antimycin A (5, 10, 25 and 50 μM) gave growth inhibition effects to THP-1 monocytes. Pyruvate and uridine were used with an intention of rescuing the THP-1 cell growth at lower antimycin A concentrations. The THP-1 monocytes treated with antimycin A with uridine and pyruvate showed more growth compared to the ones without uridine and pyruvate supplement. Yet this difference is insignificant statistically. The expressions of Hsp60 and TNF-α at the mRNA level was monitored using reverse transcriptase polymerase chain reaction. Hsp60 expression from THP-1 monocytes only showed some minor fluctuations in different antimycin A concentrations, regardless of uridine and iii pyruvate supplement, indicating the stress mitochondrial response was unobvious. On the other hand, TNF-α expression was dramatically down-regulated in THP-1 monocytes treated with antimycin A only compared to the untreated control and ones supplemented with uridine and pyruvate. These results suggest that antimycin A may have inhibition effect towards TNF-α expression, and uridine and pyruvate could also have other functions in THP-1 monocytes apart from redox rescue compounds. Yet the mitochondrial stress response shown by Hsp60 induction still remains to be further investigated.
      Date
      2009
      Type
      Thesis
      Degree Name
      Master of Science (MSc)
      Publisher
      The University of Waikato
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