Glycogen synthase kinase-3 beta inhibitors protectagainst the acute lung injuries resulting from acute necrotizing pancreatitis

Jin, Hongzhong; Yang, Xiaojia; Zhao, Kailiang; Zhao, Liang; Chen, Chen; Yu, Jia

doi:10.1590/s0102-865020190060000009

Hongzhong Jin

PhD, Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei Province, China. Acquisiton and analysis of data, manuscript writing.

* Contributed equally

https://orcid.org/0000-0002-6151-7041

Xiaojia Yang

PhD, Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei Province, China. Acquisiton and analysis of data, manuscript writing.

* Contributed equally

https://orcid.org/0000-0002-7634-3929

Kailiang Zhao

MD, PhD, Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei Province, China. Conception and design of the study, supervised all phases of the study, final approval.

https://orcid.org/0000-0003-3842-3140

Liang Zhao

MD, PhD, Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei Province, China. Statistical analysis, manuscript preparation.

https://orcid.org/0000-0003-2611-8464

Chen Chen

MD, PhD, Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei Province, China. Statistical analysis, manuscript preparation.

https://orcid.org/0000-0001-9832-6647

Jia Yu

MD, PhD, Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei Province, China. Technical procedures, histopathological examinations.

https://orcid.org/0000-0002-2503-0025

Correspondence: Kailiang Zhao. Department of Hepatobiliary Surgery. Renmin Hospital, Wuhan University. 238 Jiefang Road, Wuhan 430060 China. Phone: 86-27-88041911-81085. chensi271@aliyun.com. zhaokl1983@qq.com

*
Contributed equally

Conﬂict of interest: none

Figures (3)
Tables (3)

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Figure 1
Morphologic changes in the lung and pancreas at 12 hours after sodium taurocholate induced acute necrotizing pancreatitis. No histological alterations were observed in the pancreatic and lung tissues obtained from sham-vehicle, sham-TDZD-8 and sham-SB216763 rats. TDZD-8 and SB216763 pre-treatment significantly reduced the extent and severity of the histological signs of pancreatic and lung injury. This figure represents at least 3 experiments performed on different experimental days (original magnification, ×200).

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Figure 2
Immunohistochemical localization of NF-κB p65 in the pancreas and lung following ANP. A. Weak immunoreactivity in the cytoplasm was observed in pancreatic and lung sections obtained from sham-vehicle, sham-TDZD-8 and sham-SB216763 rats. In contrast, intense positive staining in the nucleus was observed in the pancreatic and lung sections obtained from ANP-vehicle group rats. The intensity of the positive staining in the nucleus for NF-κB p65 was markedly reduced after the administration of TDZD-8 or SB216763. But the expression of NF-κB p65 in the ANP-SB216763 group was slightly stronger than that in the ANP-TDZD-8 group. The figures represent at least three experiments performed on different experimental days (original magnification ×400). B. Immunohistochemical analysis of NF-κB p65 in pancreatic tissue. Results are presented as the mean ± standard deviation. C. Immunohistochemical analysis of NF-κB p65 in lung tissue. Results are presented as the mean ± standard deviation. Statistical significance was determined using Student’s two-tailed t-test. *P < 0.05, **P < 0.01, ***P < 0.001. ****P < 0.0001. (IOD, integrated optical density.)

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Figure 3
A. The activation of phospho-GSK-3β (Ser9), GSK-3β, iNOS, ICAM-1, IL-10 and TNF-α in lung tissue was analyzed by western blot at 12 hours following ANP. ANP markedly increased the tissue concentration of iNOS, ICAM-1 and TNF-α compared with the sham-vehicle rats. TDZD-8 or SB216763 pretreatment effectively inhibited iNOS, ICAM-1 and TNF-α activity in the lung after ANP. However, the expression of IL-10 was up-regulated in lung pretreatment with TDZD-8 or SB216763. TDZD-8 or SB216763 pretreatment induced a signiﬁcant increase in phospho-GSK-3β (Ser9). The expression of phospho-GSK-3β (ser9) in the ANP-TDZD-8 group was slightly stronger than that in the ANP-SB216763 group. Comparing the ANP-TDZD-8 group with the ANP-SB216763 group, it was found that except for TNF-α, the expression of iNOS, ICAM-1 in the ANP-TDZD-8 group was lower than that in the ANP-SB216763 group. B, C, D, E. Western blots were quantitated for iNOS, ICAM-1, IL-10 and TNF-α to β-action. F. Western blots were quantitated for phospho-GSK-3β (Ser9) to GSK-3β. The results are expressed as a fold-increase over sham-vehicle in at least 3 independent experiments. Results are presented as the mean ± standard deviation. Statistical significance was determined using Student’s two-tailed t-test. *P < 0.05, **P < 0.01, ***P < 0.001. ****P < 0.0001.

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Table 1
Experimental groups.

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Table 2
Detection of pancreas and lung function indexes in each group of rats.

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Table 3
Pancreatic and pulmonary histological score, plasma levels of IL-1β and IL-6 in each group of rats.

Groups	n	Models	Operations	Pretreatment
Sham-vehicle	12	Sham	saline (1ml/Kg)	10% DMSO
ANP-vehicle	12	ANP	5% sodium taurocholate (1ml/Kg)	10% DMSO
Sham-TDZD-8	12	Sham	saline (1ml/Kg)	TDZD-8 (1mg/Kg)
ANP-TDZD-8	12	ANP	5% sodium taurocholate (1ml/Kg)	TDZD-8 (1mg/Kg)
Sham-SB216763	12	Sham	saline (1ml/Kg)	SB216763 (1mg/Kg)
ANP-SB216763	12	ANP	5% sodium taurocholate (1ml/Kg)	SB216763 (1mg/Kg)

Groups	n	AMY(U/L)	LIPA(U/L)	Hydrothorax (g)	Lung(W/D) Ratio
Sham-vehicle	12	1560±66.11	48.3±4.59	0.20±0.04	1.52±0.06
ANP-vehicle	12	10073±343.10^a	2375±51.14^a	6.83±0.49^a	3.35±0.15^a
Sham-TDZD-8	12	1502±88.25	52.7±5.77	0.21±0.03	1.37±0.04
ANP-TDZD-8	12	4018±195.60^abc	848.6±90.64^abc	3.05±0.39^abc	2.24±0.09^abc
Sham-SB216763	12	1442±69.94	56.2±4.75	0.22±0.05	1.42±0.07
ANP-SB216763	12	5272±133.40^abde	964.6±88.79^abde	3.42±0.33^abe	2.59±0.08^abde

Groups	n	Pancreatic histological score	Pulmonary histological score	IL-1β (ng/L)	IL-6 (ng/L)
Sham-vehicle	12	0.42 ± 0.15	0.38 ± 0.15	55.7 ± 3.09	95.8 ± 1.03
ANP-vehicle	12	12.08 ± 0.30^a	9.00 ± 0.82^a	299.1 ± 15.46^a	385.4 ± 13.92^a
Sham-TDZD-8	12	0.43 ± 0.28	0.42 ± 0.15	55.4 ± 2.82	94.8 ± 0.91
ANP-TDZD-8	12	5.92± 0.39^abc	5.37 ± 0.52^abc	194.7 ± 12.17^abc	180.0 ± 10.18^abc
Sham-SB216763	12	0.50 ± 0.18	0.45 ± 0.18	51.2 ± 2.63	99.7 ± 2.03
ANP-SB216763	12	6.75 ± 0.38^abde	7.42 ± 0.62^abde	220.0 ± 17.70^abde	212.6 ± 9.32^abde

[1] Correspondence: Kailiang Zhao. Department of Hepatobiliary Surgery. Renmin Hospital, Wuhan University. 238 Jiefang Road, Wuhan 430060 China. Phone: 86-27-88041911-81085. chensi271@aliyun.com. zhaokl1983@qq.com

Brasil

Brasil

Glycogen synthase kinase-3 beta inhibitors protectagainst the acute lung injuries resulting from acute necrotizing pancreatitis¹ 1 Research performed at the Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei, China.

Abstract

Purpose

Methods

Results

Conclusions

Brasil

Brasil

Glycogen synthase kinase-3 beta inhibitors protectagainst the acute lung injuries resulting from acute necrotizing pancreatitis1 1 Research performed at the Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei, China.

Abstract

Purpose

Methods

Results

Conclusions

Glycogen synthase kinase-3 beta inhibitors protectagainst the acute lung injuries resulting from acute necrotizing pancreatitis¹ 1 Research performed at the Department of Hepatobiliary Surgery, Renmin Hospital, Wuhan University, Hubei, China.