Heat Shock Protein 70 Messenger RNA in Rat Leukocytes Elevates After Severe Intestinal Ischemia–Reperfusion

• Published in: The Journal of surgical research 2019-10, Vol.242, p.342-348
• Main Authors: Mine, Yuka, Fujita, Fumihiko, Murase, Takehiko, Ito, Shinichiro, Takatsuki, Mitsuhisa, Ikematsu, Kazuya, Eguchi, Susumu
• Format: Article
• Language: English
• Subjects: Animals; Biomarkers - blood; Biomarkers - metabolism; Disease Models, Animal; Heat shock protein 70 (HSP70); HSP70 Heat-Shock Proteins - blood; HSP70 Heat-Shock Proteins - genetics; HSP70 Heat-Shock Proteins - metabolism; Humans; Intestinal Mucosa - blood supply; Intestinal Mucosa - pathology; Intestine, Small - blood supply; Intestine, Small - pathology; Ischemia–reperfusion injury (IRI); Leukocytes; Leukocytes - metabolism; Male; Mesenteric Artery, Superior - surgery; Mesenteric Ischemia - blood; Mesenteric Ischemia - etiology; Mesenteric Ischemia - pathology; Rat small intestine; Rats; Rats, Wistar; Reperfusion Injury - blood; Reperfusion Injury - etiology; Reperfusion Injury - pathology; RNA, Messenger - blood; RNA, Messenger - metabolism
• ISSN: 0022-4804; 1095-8673
• DOI: 10.1016/j.jss.2019.04.074

Heat shock protein 70 (HSP70) confers protection against heat shock, oxidative stress, infection, and inflammation in many cell types. A recent study reported that the induction of HSP70 was associated with morphologic protection against ischemia–reperfusion injury (IRI) in the rat small intestine. This study investigated the dynamics of HSP70 in leukocytes during intestinal IRI in a rat model. Serial blood samples were collected at 60-minute intervals up to 240 min from male Wistar rats (n = 15). The rats were divided into three groups of five each: the control group, the nonlethal IRI group, and the lethal IRI group. Rats belonging to the control group underwent a sham operation, and laparotomy was performed on rats in the lethal and nonlethal IRI groups. The nonlethal group experienced a 30-minute clamping of the superior mesenteric artery, and the lethal group experienced a 75-minute clamping of the superior mesenteric artery. The expression of HSP70 messenger RNA (mRNA) in leukocytes was measured by real-time quantitative polymerase chain reaction. Mixed-effects modeling of repeated measures was used to carry out the statistical analysis. The Bonferroni correction was applied to multiple comparisons. A P value < 0.0167 was considered to indicate statistical significance. The expression of HSP70 mRNA in leukocytes increased 60 min after reperfusion in both IRI groups, and it was 12.8 times higher in the lethal group and 3.6 times higher in the nonlethal group compared with the control group. The expression of mRNA in the lethal group was significantly increased compared with the nonlethal group and the control group at 120 and 180 min after reperfusion. At 120 min after reperfusion, the expression of HSP70 mRNA was 6.1 times higher in the lethal group than in the nonlethal group (P = 0.0075) and 17.7 times higher than in the control group (P = 0.0011). At 180 min after reperfusion, the expression of HSP70 mRNA was 6.8 times higher in the lethal group than in the nonlethal group (P = 0.0007) and 4.3 times higher than in the control group (P = 0.0032). Although the expression of HSP70 mRNA in the nonlethal group was elevated in the early stages of reperfusion, there was no difference between the nonlethal group and the control group (P = 0.0212 at 60 min). The expression of HSP70 mRNA in leukocytes may be a clinically useful indicator for evaluating pathologic conditions in intestinal IRI.