Page 57 - GTM-3-2

Global Translational Medicine Hesperetin alleviates pulmonary injury

1. Introduction investigate the effect of hesperetin administration on
pulmonary tissue architecture and immunohistochemical
Pulmonary contusion (PC) is a common injury due to blunt expression of pulmonary iNOS. Further, we examined the
chest trauma observed in 30 – 75% of patients attending effect of hesperetin on serum inflammatory markers such
emergency medical and trauma centers. In particular, as IL-1β, IL-6, and TNF-α and the oxidative stress markers
1
high rates of PC were observed after the occurrence of such as superoxide dismutase (SOD), malondialdehyde
chest injuries in pediatric patients. Regardless of the (MDA), and catalase (CAT) enzyme levels in a rat PC
2
severity, estimated rates of mortality attributable to PC model.
range from 14% to 40%. In addition, it has been reported
as the direct cause of death in 20 – 25% of patients with 2. Materials and methods
multiple trauma. PC may trigger acute pulmonary injury,
3
4
pneumonia, and respiratory failure syndromes. In addition, 2.1. Chemicals
hypoxia caused by PC may affect functions of distant organ The hesperetin used in this research was obtained from
such as kidneys, brain, and liver. Although the definite Santa Cruz (Santa Cruz Biotech, Dallas, TX). Hematoxylin
5
mechanisms of PC have not been fully clarified, factors and eosin were obtained from Merck (Germany). Ketamine
such as increased pulmonary permeability and edema, was purchased from Pfizer (İstanbul, Turkey) and xylazine
inflammation, and ventilation/perfusion incompatibility was from Bayer (Turkiye). iNOS primary antibodies
have been reported to play roles in its pathophysiology. were purchased by Novus Biologicals (Littleton, USA).
6,7
While there is no specific pharmacological treatment for IL-1β, IL-6, and TNF-α ELISA kits were obtained from
PC, symptomatic and supportive treatment are applicable Shanghai Biotech (Shanghai YL Biotech Co., Shanghai,
to the affected patients. 6 China). The MDA kit was acquired from Nanjing
PC may develop along with abnormal inflammatory Jiancheng Bioengineering Institute (Nanjing, China).
response and heightened oxidative stress following blunt Secondary antibody kits used for immunohistochemical
chest trauma. Chest trauma can trigger the activation of labeling (Thermo Scientific, Fremont, CA, USA) and AEC
2
crucial transcription factors, such as nuclear factor kappa (3-amino-9-ethylcarbazole) used as chromogen were
beta, which have a pivotal role in inflammation and the obtained from Thermo Scientific (USA).
transcription of proinflammatory genes, leading to the
activation and synthesis of pro-inflammatory cytokines 2.2. Animals
such as interleukin (IL)-1β, IL-6 and inducible nitric Animals used in this study were obtained from Tekirdağ
7
oxide synthase (iNOS). The pulmonary inflammation Namık Kemal University Experimental Animals Research
and tissue injury occurring with PC has been reported to and Implementation Center (NKUDAM). Eighteen male
be associated with increased pro-inflammatory cytokine Wistar albino rats (250 – 300 g, 8 – 10 weeks old) were
levels in many studies. 8,9 used in this study. During the study duration, animals were
Experimental investigations have found that certain housed at NKUDAM under standard conditions (22 ± 2°C
pharmacological compounds with anti-inflammatory and temperature, 50 – 60% humidity, and 12-h light/12-h dark
antioxidative properties can suppress pro-inflammatory cycle) and fed standard pellet feed and water ad libitum.
cytokine production and oxidative stress, alleviating 2.3. Ethical approval
tissue damage caused by PC and improving clinical
symptoms. 6,10,11 Hesperetin is a natural bioflavonoid All procedures in this study were approved by Tekirdağ
derivative synthesized after the hydrolysis of hesperidin. Namık Kemal University Experimental Animals Local
Among its many pharmacological effects, it is known to be Ethics Committee. All experimental procedures in the
antioxidant, anti-inflammatory, antiapoptotic, antitumor, study were completed in accordance with the Care and Use
and antihyperlipidemic. Administration of hesperetin of Laboratory Animals Guidelines recommended by the
12
has been reported to support anti-inflammatory effects and National Institutes of Health.
act as a strong free radical scavenger in many in vivo and 2.4. Experimental procedures
in vitro experimental studies. 13,14 In a lipopolysaccharide
(LPS)-induced pulmonary injury model, hesperetin The rats used in this study were divided into three groups
administration was found to reduce tumor necrosis factor (n = 6 each): control, PC, and PC + hesperetin group.
alpha (TNF-α) and serum IL-6 levels and protect tissue No intervention was performed to the control group.
structure. However, the protective role of hesperetin PC induction was executed according to a previously
15
therapy in the model of blunt chest trauma-induced PC reported method. The right hemithorax of each rat under
16
has not yet been investigated. Therefore, we aimed to anesthesia (ketamine-xylazine; 50 – 15 mg/kg, ip), which

Volume 3 Issue 2 (2024) 2 doi: 10.36922/gtm.2568