Korean J Physiol Pharmacol.  2021 Jul;25(4):281-296. 10.4196/kjpp.2021.25.4.281.

Whole body hypoxic preconditioning-mediated multiorgan protection in db/db mice via nitric oxide-BDNF-GSK-3β-Nrf2 signaling pathway

• Li Y
• Huang Y
• Cheng X
• He Y
• Hu X

• KMID: 2516868
• DOI: http://doi.org/10.4196/kjpp.2021.25.4.281

Abstract

The beneficial effects of hypoxic preconditioning are abolished in the diabetes. The present study was designed to investigate the protective effects and mechanisms of repeated episodes of whole body hypoxic preconditioning (WBHP) in db/db mice. The protective effects of preconditioning were explored on diabetesinduced vascular dysfunction, cognitive impairment and ischemia-reperfusion (IR)-induced increase in myocardial injury. Sixteen-week old db/db (diabetic) and C57BL/6 (non-diabetic) mice were employed. There was a significant impairment in cognitive function (Morris Water Maze test), endothelial function (acetylcholineinduced relaxation in aortic rings) and a significant increase in IR-induced heart injury (Langendorff apparatus) in db/db mice. WBHP stimulus was given by exposing mice to four alternate cycles of low (8%) and normal air O₂ for 10 min each. A single episode of WBHP failed to produce protection; however, two and three episodes of WBHP significantly produced beneficial effects on the heart, brain and blood vessels. There was a significant increase in the levels of brain-derived neurotrophic factor (BDNF) and nitric oxide (NO) in response to 3 episodes of WBHP. Moreover, pretreatment with the BDNF receptor, TrkB antagonist (ANA-12) and NO synthase inhibitor (LNAME) attenuated the protective effects imparted by three episodes of WBHP. These pharmacological agents abolished WBHP-induced restoration of p-GSK-3β/GSK-3β ratio and Nrf2 levels in IR-subjected hearts. It is concluded that repeated episodes of WHBP attenuate cognitive impairment, vascular dysfunction and enhancement in IRinduced myocardial injury in diabetic mice be due to increase in NO and BDNF levels that may eventually activate GSK-3β and Nrf2 signaling pathway to confer protection.

Keyword

Brain-derived neurotrophic factor; Dementia; Heart; Ischemia; Myocardial infarction; Nitric oxide

Figure

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  Fig. 1 Effects of different interventions on the extent of myocardial injury assessed by measuring the release of specific marker of heart injury i.e., creatine kinase (CK)-MB in the coronary effluent. The measurement was done before subjecting the heart to ischemia (before ischemia) and at the start of reperfusion. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. basal; bp < 0.05 vs. normal; cp < 0.05 vs. diabetes; dp < 0.05 vs. 1 HP in normal; ep < 0.05 vs. 3 HP in diabetes.

• 

  Fig. 2 Effects of different interventions on the extent of myocardial injury assessed by measuring the release of specific marker of heart injury i.e., cardiac troponin T (cTnT) in the coronary effluent. The measurement was done before subjecting the heart to ischemia (before ischemia) and at the start of reperfusion. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. basal; bp < 0.05 vs. normal; cp < 0.05 vs. diabetes; dp < 0.05 vs. 1 HP in normal; ep < 0.05 vs. 3 HP in diabetes.

• 

  Fig. 3 Effects of different interventions on the extent of myocardial injury assessed by measuring the release of specific marker of heart injury i.e., lactate dehydrogenase-1 (LDH-1) in the coronary effluent. The measurement was done before subjecting the heart to ischemia (before ischemia) and at the start of reperfusion. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. basal; bp < 0.05 vs. normal; cp < 0.05 vs. diabetes; dp < 0.05 vs. 1 HP in normal; ep < 0.05 vs. 3 HP in diabetes.

• 

  Fig. 4 Effects of different interventions on the markers of apoptosis i.e., caspase 3 activity and Bcl-2 expression in the heart homogenates after ischemia-reperfusion injury. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. normal; bp < 0.05 vs. diabetes; cp < 0.05 vs. 1 HP in normal; dp < 0.05 vs. 3 HP in diabetes.

• 

  Fig. 5 Effects of different interventions on the memory retrieval (how much information is retained following learning) in the form of time spent in target quadrant (TSTQ). It was assessed on the 5th day of trial in the Morris Water Maze test. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. normal; bp < 0.05 vs. diabetes; cp < 0.05 vs. 3 HP in diabetes.

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  Fig. 6 Effects of different interventions on the plasma glucose levels in the normal and diabetic mice. The levels were measured at the end of experiment. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. normal.

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  Fig. 7 Effects of different interventions on the nitric oxide, which was indirectly assessed by measuring the levels of nitrite in the serum. The levels of serum nitrite were measured 1 h after hypoxic preconditioning stimulus. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. normal; bp < 0.05 vs. diabetes; cp < 0.05 vs. 1 HP in normal; dp < 0.05 vs. 3 HP in diabetes.

• 

  Fig. 8 Effects of different interventions on the pharmacological target employed in this study i.e., brain-derived neurotrophic factor (BDNF). The levels of BDNF were measured in the serum 1 h after hypoxic preconditioning stimulus. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. normal; bp < 0.05 vs. diabetes; cp < 0.05 vs. 1 HP in normal; dp < 0.05 vs. 3 HP in diabetes.

• 

  Fig. 9 Effects of different interventions on intracellular signaling pathway i.e., p-GSK-3β/GSK-3β ratio and Nrf2 expression in heart homogenates following ischemia-reperfusion injury. HP, hypoxic preconditioning; 1 HP, single episode of hypoxic preconditioning. ap < 0.05 vs. control; bp < 0.05 vs. normal; cp < 0.05 vs. diabetes; dp < 0.05 vs. 1 HP in normal; ep < 0.05 vs. 3 HP in diabetes.

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