Pyruvate Dehydrogenase and Tricarboxylic Acid Cycle Enzymes Are Sensitive Targets of Traumatic Brain Injury Induced Metabolic Derangement.

Giacomo Lazzarino,Angela Maria Amorini,Antonio Belli,Valentina Di Pietro,Stefano Signoretti,Francesco Saverio Pastore,Giuseppe Lazzarino,Giuseppe Caruso,Giuseppe Musumeci,Barbara Tavazzi

doi:10.3390/ijms20225774

Abstract

Using a closed-head impact acceleration model of mild or severe traumatic brain injury (mTBI or sTBI, respectively) in rats, we evaluated the effects of graded head impacts on the gene and protein expressions of pyruvate dehydrogenase (PDH), as well as major enzymes of mitochondrial tricarboxylic acid cycle (TCA). TBI was induced in anaesthetized rats by dropping 450 g from 1 (mTBI) or 2 m height (sTBI). After 6 h, 12 h, 24 h, 48 h, and 120 h gene expressions of enzymes and subunits of PDH. PDH kinases and phosphatases (PDK1-4 and PDP1-2, respectively), citrate synthase (CS), isocitrate dehydrogenase (IDH), oxoglutarate dehydrogenase (OGDH), succinate dehydrogenase (SDH), succinyl-CoA synthase (SUCLG), and malate dehydrogenase (MDH) were determined in whole brain extracts (n = 6 rats at each time for both TBI levels). In the same samples, the high performance liquid chromatographic (HPLC) determination of acetyl-coenzyme A (acetyl-CoA) and free coenzyme A (CoA-SH) was performed. Sham-operated animals (n = 6) were used as controls. After mTBI, the results indicated a general transient decrease, followed by significant increases, in PDH and TCA gene expressions. Conversely, permanent PDH and TCA downregulation occurred following sTBI. The inhibitory conditions of PDH (caused by PDP1-2 downregulations and PDK1-4 overexpression) and SDH appeared to operate only after sTBI. This produced almost no change in acetyl-CoA and free CoA-SH following mTBI and a remarkable depletion of both compounds after sTBI. These results again demonstrated temporary or steady mitochondrial malfunctioning, causing minimal or profound modifications to energy-related metabolites, following mTBI or sTBI, respectively. Additionally, PDH and SDH appeared to be highly sensitive to traumatic insults and are deeply involved in mitochondrial-related energy metabolism imbalance.

Highlights

Mitochondria are the key organelles regulating cellular bioenergetic and redox balance and apoptosis [1]
Using a closed-head impact acceleration model of mild or severe traumatic brain injury in rats, we evaluated the effects of graded head impacts on the gene and protein expressions of pyruvate dehydrogenase (PDH), as well as major enzymes of mitochondrial tricarboxylic acid cycle (TCA)
The results of the present study indicate that, depending on the severity of the traumatic insult, the PDH complex and enzymes of the TCA cycle are sensitive targets of traumatic brain injury (TBI) pathology

Summary

Introduction

Mitochondria are the key organelles regulating cellular bioenergetic and redox balance and apoptosis [1]. The main mitochondrial function is to provide adequate energy production via ATP. The pathological decrease in this activity negatively affects cellular energy-dependent reactions, which deeply influences other mitochondrial functions and leads to a decrease in cell survival. ATP synthesis occurs in a stoichiometric ratio, linking the flow of electrons through ETC To water (namely, oxygen consumption) and, the amount of protons (n = 10 H+) transferred into the mitochondrial inner membrane space during the electron flow. The maintenance of the aforementioned stoichiometry allows the synthesis of 2.5-3 or 1.5-2 ATP when the electron donors are, respectively, NADH or FADH2. ATP production through oxidative phosphorylation (OXPHOS) is strictly dependent on the correct ETC functioning and electroosmotic gradient formation between the intermembrane space and the mitochondrial matrix

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Results

Conclusion