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Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA.
J Surg Res. 2012 Aug; 176(2):614-20.JS

Abstract

BACKGROUND

Tumor necrosis factor-alpha (TNF-α) is a central mediator in the hepatic response to ischemia/reperfusion. Short hairpin RNA (shRNA) has been proven to be an effective means of harnessing the RNA interference pathway in mammalian cells. In the current study, we investigated whether silencing TNF-α gene with shRNA can prevent liver ischemic reperfusion injury (IRI).

METHODS

Male BalB/c mice were randomized to TNF-α shRNA, scramble shRNA, or sham operation groups. TNF-α shRNA and scramble shRNA groups were injected 48 h before inducing IRI. IRI was induced via microaneurysm clamps applied to the left hepatic artery and portal vein. Six hours after reperfusion, IRI injury was examined by serum level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), liver histopathology, MPO, and MDA level, as well as by relative quantities of TNF-α mRNA.

RESULTS

TNF-α expression induced by ischemia reperfusion in the liver was significantly suppressed after treatment with TNF-α shRNA compared with the group treated with scramble shRNA (P < 0.001). Mice treated with TNF-α shRNA showed lower peak values of AST and ALT than scramble shRNA treated mice (P < 0.001). On histopathologic slides, mice treated with TNF-α shRNA had significantly less ischemia/reperfusion injury based on Suzuki score than the scramble shRNA group, 3.57 ± 2.30 and 8.83 ± 0.98 respectively (P < 0.001), while the sham group was not significantly different from the TNF-alpha shRNA group, 0 ± 0 and 3.57 ± 2.30, respectively (P = 0.075). Liver tissue MDA levels were significantly lower in mice treated with TNF-α shRNA as compared with the group treated with scramble shRNA (P < 0.01). Immunohistochemical staining for MPO was significantly lower in mice treated with TNF-α shRNA compared with the group treated with shRNA (compared with treated with scramble shRNA group.)

CONCLUSIONS

Liver IRI can be minimized through gene silencing of TNF-α. This may represent a novel therapy in the setting of transplantation and in other conditions associated with IRI of the liver.

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Authors+Show Affiliations

Hernandez-Alejandro R
Multi-Organ Transplant Program, London Health Sciences Centre, The University of Western Ontario, London, Canada. roberto.hernandezalejandro@lhsc.on.ca
Zhang X
No affiliation info available
Croome KP
No affiliation info available
Zheng X
No affiliation info available
Parfitt J
No affiliation info available
Chen D
No affiliation info available
Jevnikar A
No affiliation info available
Wall W
No affiliation info available
Min WP
No affiliation info available
Quan D
No affiliation info available

MeSH

Alanine TransaminaseAnimalsAspartate AminotransferasesDown-RegulationGenetic TherapyLiverMaleMalondialdehydeMiceMice, Inbred BALB CPeroxidaseRNA, Small InterferingReperfusion InjuryTumor Necrosis Factor-alpha

Pub Type(s)

Journal Article

Language

eng

PubMed ID

22221603

Citation

Hernandez-Alejandro, Roberto, et al. "Reduction of Liver Ischemia Reperfusion Injury By Silencing of TNF-α Gene With ShRNA." The Journal of Surgical Research, vol. 176, no. 2, 2012, pp. 614-20.
Hernandez-Alejandro R, Zhang X, Croome KP, et al. Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. J Surg Res. 2012;176(2):614-20.
Hernandez-Alejandro, R., Zhang, X., Croome, K. P., Zheng, X., Parfitt, J., Chen, D., Jevnikar, A., Wall, W., Min, W. P., & Quan, D. (2012). Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. The Journal of Surgical Research, 176(2), 614-20. https://doi.org/10.1016/j.jss.2011.10.004
Hernandez-Alejandro R, et al. Reduction of Liver Ischemia Reperfusion Injury By Silencing of TNF-α Gene With ShRNA. J Surg Res. 2012;176(2):614-20. PubMed PMID: 22221603.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. AU - Hernandez-Alejandro,Roberto, AU - Zhang,Xusheng, AU - Croome,Kris P, AU - Zheng,Xiufen, AU - Parfitt,Jeremy, AU - Chen,Dong, AU - Jevnikar,Anthony, AU - Wall,William, AU - Min,Wei-Ping, AU - Quan,Douglas, Y1 - 2011/11/01/ PY - 2011/07/24/received PY - 2011/09/23/revised PY - 2011/10/06/accepted PY - 2012/1/7/entrez PY - 2012/1/10/pubmed PY - 2012/10/4/medline SP - 614 EP - 20 JF - The Journal of surgical research JO - J Surg Res VL - 176 IS - 2 N2 - BACKGROUND: Tumor necrosis factor-alpha (TNF-α) is a central mediator in the hepatic response to ischemia/reperfusion. Short hairpin RNA (shRNA) has been proven to be an effective means of harnessing the RNA interference pathway in mammalian cells. In the current study, we investigated whether silencing TNF-α gene with shRNA can prevent liver ischemic reperfusion injury (IRI). METHODS: Male BalB/c mice were randomized to TNF-α shRNA, scramble shRNA, or sham operation groups. TNF-α shRNA and scramble shRNA groups were injected 48 h before inducing IRI. IRI was induced via microaneurysm clamps applied to the left hepatic artery and portal vein. Six hours after reperfusion, IRI injury was examined by serum level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), liver histopathology, MPO, and MDA level, as well as by relative quantities of TNF-α mRNA. RESULTS: TNF-α expression induced by ischemia reperfusion in the liver was significantly suppressed after treatment with TNF-α shRNA compared with the group treated with scramble shRNA (P < 0.001). Mice treated with TNF-α shRNA showed lower peak values of AST and ALT than scramble shRNA treated mice (P < 0.001). On histopathologic slides, mice treated with TNF-α shRNA had significantly less ischemia/reperfusion injury based on Suzuki score than the scramble shRNA group, 3.57 ± 2.30 and 8.83 ± 0.98 respectively (P < 0.001), while the sham group was not significantly different from the TNF-alpha shRNA group, 0 ± 0 and 3.57 ± 2.30, respectively (P = 0.075). Liver tissue MDA levels were significantly lower in mice treated with TNF-α shRNA as compared with the group treated with scramble shRNA (P < 0.01). Immunohistochemical staining for MPO was significantly lower in mice treated with TNF-α shRNA compared with the group treated with shRNA (compared with treated with scramble shRNA group.) CONCLUSIONS: Liver IRI can be minimized through gene silencing of TNF-α. This may represent a novel therapy in the setting of transplantation and in other conditions associated with IRI of the liver. SN - 1095-8673 UR - https://www.unboundmedicine.com/medline/citation/22221603/Reduction_of_liver_ischemia_reperfusion_injury_by_silencing_of_TNF_��_gene_with_shRNA_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-4804(11)00834-1 DB - PRIME DP - Unbound Medicine ER -