J Korean Med Sci.  2014 Apr;29(4):556-563. 10.3346/jkms.2014.29.4.556.

Effects of Transglutaminase 2 Inhibition on Ventilator-Induced Lung Injury

• Suh IB
• Yoon DW
• Oh WO
• Lee EJ
• Min KH
• Hur GY
• Lee SH
• Lee SY
• Lee SY
• Shin C
• Shim JJ
• In KH
• Kang KH
• Kim JH

• KMID: 1774462
• DOI: http://doi.org/10.3346/jkms.2014.29.4.556

Abstract

This study was performed to examine the role of transglutaminase 2 (TG2) in ventilator-induced lung injury (VILI). C57BL/6 mice were divided into six experimental groups: 1) control group; 2) lipopolysaccharide (LPS) group; 3) lung protective ventilation (LPV) group; 4) VILI group; 5) VILI with cystamine, a TG2 inhibitor, pretreatment (Cyst+VILI) group; and 6) LPV with cystamine pretreatment (Cyst+LPV) group. Acute lung injury (ALI) score, TG2 activity and gene expression, inflammatory cytokines, and nuclear factor-kappaB (NF-kappaB) activity were measured. TG2 activity and gene expression were significantly increased in the VILI group (P < 0.05). Cystamine pretreatment significantly decreased TG2 activity and gene expression in the Cyst+VILI group (P < 0.05). Inflammatory cytokines were higher in the VILI group than in the LPS and LPV groups (P < 0.05), and significantly lower in the Cyst+VILI group than the VILI group (P < 0.05). NF-kappaB activity was increased in the VILI group compared with the LPS and LPV groups (P < 0.05), and significantly decreased in the Cyst+VILI group compared to the VILI group (P = 0.029). The ALI score of the Cyst+VILI group was lower than the VILI group, but the difference was not statistically significant (P = 0.105). These results suggest potential roles of TG2 in the pathogenesis of VILI.

Keyword

Acute Lung Injury; Respiratory Distress Syndrome, Adult; Respiration, Artificial; Ventilator-Induced Lung Injury; Inflammation; Transglutaminase 2

MeSH Terms

Acute Lung Injury/pathology
Animals
Cystamine/therapeutic use
Cytokines/analysis
Enzyme Inhibitors/therapeutic use
Enzyme-Linked Immunosorbent Assay
GTP-Binding Proteins/*antagonists & inhibitors/genetics/metabolism
Gene Expression
Lipopolysaccharides/toxicity
Male
Mice
Mice, Inbred C57BL
NF-kappa B/metabolism
Respiration, Artificial
Transglutaminases/*antagonists & inhibitors/genetics/metabolism
Ventilator-Induced Lung Injury/*enzymology/pathology/prevention & control
Cystamine
Cytokines
Enzyme Inhibitors
GTP-Binding Proteins
Lipopolysaccharides
NF-kappa B
Transglutaminases

Figure

• 

  Fig. 1 Determination pretreatment time, transglutaminase 2 (TG2) activity and gene expression. To determine the optimal pretreatment time, cystamine was administered intraperitoneally to six mice at 36, 24, 12, 6, 4, 2, and 0 hr before LPS instillation and 4 hr of MV. The lowest TG2 activity and TG2 RT-PCR are observed at the 6-hr cystamine pretreatment time (P = 0.001 by Kruskal-Wallis test, *P < 0.05, compared with the VILI and other time points) (A). TG2 activity (B) and gene expression (C) are significantly higher in the VILI group than in other groups (P < 0.05). In the Cyst+VILI group, TG2 activity and gene expression are lower than in the VILI group (*P = 0.029 and **P = 0.016, respectively). Cystamine pretreatment has no additional effect on TG2 activity and gene expression in the Cyst+LPV group comparing with the LPV group (†P = 0.114 and ‡P = 0.19, respectively).

• 

  Fig. 2 Inflammatory cytokines and nuclear factor-κB (NF-κB) activity. The concentration of tumor necrosis factor-α (TNF-α) (A), interleukin (IL)-1β (B), and IL-6 (C) and NF-κB activity (D) are significantly increased in the VILI group compared to other groups (P < 0.05). The Cyst+VILI group demonstrates significantly decreased inflammatory cytokine levels and NF-κB activity than the VILI group (*P = 0.016, **P = 0.026, ***P = 0.015, and §P = 0.029, respectively). In the comparisons between the LPV and Cyst+LPV groups, the inflammatory cytokine levels and NF-κB activity are not different (†P = 1.000, ‡‡P = 0.19, ‡P = 0.429, and ‡‡P = 0.114, respectively). ND, not detectable; OD, optical density.

• 

  Fig. 3 Histopathologic findings and acute lung injury (ALI) scores. The LPS group (B) shows typical ALI findings, including intra-alveolar exudates, inflammatory cell infiltration, intra-alveolar hemorrhage, and interstitial edema that were not found in the control group (A). These findings are exacerbated in the VILI group (C). In the Cyst+VILI group (D), the degree of ALI is mildly decreased compared to the VILI group (C). The findings of LPV (E) and Cyst+LPV (F) are similar to the LPS group (B). The ALI score (G) of the VILI group is significantly higher than those of the control, LPS, LPV, and Cyst+LPV groups (*P < 0.05). The Cyst+VILI group has non-significantly lower ALI scores than the VILI group (†P = 0.105), and is not different compared to the LPS, LPV, and Cyst+LPV groups (‡P > 0.05).

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