Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA

Roberto Hernandez-Alejandro, Xusheng Zhang, Kris P. Croome, Xiufen Zheng, Jeremy Parfitt, Dong Chen, Anthony Jevnikar, William Wall, Wei Ping Min, Douglas Quan

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Pages (from-to)614-620
Number of pages7
JournalJournal of Surgical Research
Volume176
Issue number2
DOIs
StatePublished - Aug 2012
Externally publishedYes

Fingerprint

Reperfusion Injury
Small Interfering RNA
Tumor Necrosis Factor-alpha
Liver
Genes
Reperfusion
Aspartate Aminotransferases
Alanine Transaminase
Ischemia
Hepatic Veins
Hepatic Artery
Gene Silencing
Portal Vein
RNA Interference

Keywords

  • animal model
  • ischemia reperfusion injury
  • shRNA
  • TNF-α

ASJC Scopus subject areas

  • Surgery

Cite this

Hernandez-Alejandro, R., Zhang, X., Croome, K. P., Zheng, X., Parfitt, J., Chen, D., ... Quan, D. (2012). Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. Journal of Surgical Research, 176(2), 614-620. https://doi.org/10.1016/j.jss.2011.10.004

Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. / Hernandez-Alejandro, Roberto; Zhang, Xusheng; Croome, Kris P.; Zheng, Xiufen; Parfitt, Jeremy; Chen, Dong; Jevnikar, Anthony; Wall, William; Min, Wei Ping; Quan, Douglas.

In: Journal of Surgical Research, Vol. 176, No. 2, 08.2012, p. 614-620.

Research output: Contribution to journalArticle

Hernandez-Alejandro, R, Zhang, X, Croome, KP, Zheng, X, Parfitt, J, Chen, D, Jevnikar, A, Wall, W, Min, WP & Quan, D 2012, 'Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA', Journal of Surgical Research, vol. 176, no. 2, pp. 614-620. https://doi.org/10.1016/j.jss.2011.10.004
Hernandez-Alejandro R, Zhang X, Croome KP, Zheng X, Parfitt J, Chen D et al. Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. Journal of Surgical Research. 2012 Aug;176(2):614-620. https://doi.org/10.1016/j.jss.2011.10.004
Hernandez-Alejandro, Roberto ; Zhang, Xusheng ; Croome, Kris P. ; Zheng, Xiufen ; Parfitt, Jeremy ; Chen, Dong ; Jevnikar, Anthony ; Wall, William ; Min, Wei Ping ; Quan, Douglas. / Reduction of liver ischemia reperfusion injury by silencing of TNF-α gene with shRNA. In: Journal of Surgical Research. 2012 ; Vol. 176, No. 2. pp. 614-620.
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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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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

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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.

AB - 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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