Haemoxygenase modulates cytokine induced neutrophil chemoattractant in hepatic ischemia reperfusion injury.
OBJECTIVE
To investigate the hepatic microcirculatory changes due to Haemoxygenase (HO), effect of HO inhibition on remote ischemic preconditioning (RIPC) and modulation of CINC.


METHODS
Eight groups of animals were studied - Sham, ischemia reperfusion injury (IRI) the animals were subjected to 45 min of hepatic ischemia followed by three hours of reperfusion, RIPC (remote ischemic preconditioning) + IRI group, remote ischemic preconditioning in sham (RIPC + Sham), PDTC + IR (Pyridodithiocarbamate, HO donor), ZnPP + RIPC + IRI (Zinc protoporphyrin prior to preconditioning), IR-24 (45 min of ischemia followed by 24 h of reperfusion), RIPC + IR-24 (preconditioning prior to IR). After 3 and 24 h of reperfusion the animals were killed by exsanguination and samples were taken.


RESULTS
Velocity of flow (160.83 ± 12.24 μm/s), sinusoidal flow (8.42 ± 1.19) and sinusoidal perfusion index (42.12 ± 7.28) in hepatic IR were lower (P < 0.05) in comparison to RIPC and PDTC (HO inducer). RIPC increased velocity of flow (328.04 ± 19.13 μm/s), sinusoidal flow (17.75 ± 2.59) and the sinusoidal perfusion index (67.28 ± 1.82) (P < 0.05). PDTC (HO induction) reproduced the effects of RIPC in hepatic IR. PDTC restored RBC velocity (300.88 ± 22.109 μm/s), sinusoidal flow (17.66 ± 3.71) and sinusoidal perfusion (82.33 ± 3.5) to near sham levels. ZnPP (HO inhibition) reduced velocity of flow of RBC in the RIPC group (170.74 ± 13.43 μm/s and sinusoidal flow in the RIPC group (9.46 ± 1.34). ZnPP in RIPC (60.29 ± 1.82) showed a fall in perfusion only at 180 min of reperfusion. Neutrophil adhesion in IR injury is seen in both postsinusoidal venules (769.05 ± 87.48) and sinusoids (97.4 ± 7.49). Neutrophil adhesion in RIPC + IR injury is reduced in both postsinusoidal venules (219.66 ± 93.79) and sinusoids (25.69 ± 9.08) (P < 0.05). PDTC reduced neutrophil adhesion in both postsinusoidal venules (89.58 ± 58.32) and sinusoids (17.98 ± 11.01) (P < 0.05) reproducing the effects of RIPC. ZnPP (HO inhibition) increased venular (589.04 ± 144.36) and sinusoidal neutrophil adhesion in preconditioned animals (121.39 ± 30.65) (P < 0.05). IR after 24 h of reperfusion increased venular and sinusoidal neutrophil adhesion in comparison to the early phase and was significantly reduced by RIPC. Hepatocellular cell death in IRI (80.83 ± 13.03), RIPC + IR (17.35 ± 2.47), and PTDC + IR (11.66 ± 1.17) reduced hepatocellular death. ZnPP + RIPC + IR (41.33 ± 3.07) significantly increased hepatocellular death (P < 0.05 PTDC/RIPC vs ZnPP and IR). The CINC cytokine levels in sham (101.32 ± 6.42). RIPC + sham (412.18 ± 65.24) as compared to sham (P < 0.05). CINC levels in hepatic IR were (644.08 ± 181.24). PDTC and RIPC CINC levels were significantly lower than hepatic IR (P < 0.05). HO inhibition in preconditioned animals with Zinc protoporphyrin increased serum CINC levels (521.81 ± 74.9) (P < 0.05). The serum CINC levels were high in the late phase of hepatic IR (15306 ± 1222.04). RIPC reduced CINC levels in the late phase of IR (467.46 ± 26.06), P < 0.05.


CONCLUSIONS
RIPC protects hepatic microcirculation by induction of HO and modulation of CINC in hepatic IR.
Niteen Tapuria, Sameer Junnarkar, Mahmoud Abu-amara, Barry Fuller, Alexander M Seifalian, Brian R Davidson, Department of HPB Surgery and Liver transplantation, Royal Free Hospital, London NW32QG, United Kingdom.
Niteen
N
Tapuria
Niteen Tapuria, Sameer Junnarkar, Mahmoud Abu-amara, Barry Fuller, Alexander M Seifalian, Brian R Davidson, Department of HPB Surgery and Liver transplantation, Royal Free Hospital, London NW32QG, United Kingdom.
Sameer
S
Junnarkar
Niteen Tapuria, Sameer Junnarkar, Mahmoud Abu-amara, Barry Fuller, Alexander M Seifalian, Brian R Davidson, Department of HPB Surgery and Liver transplantation, Royal Free Hospital, London NW32QG, United Kingdom.
Mahmoud
M
Abu-Amara
Niteen Tapuria, Sameer Junnarkar, Mahmoud Abu-amara, Barry Fuller, Alexander M Seifalian, Brian R Davidson, Department of HPB Surgery and Liver transplantation, Royal Free Hospital, London NW32QG, United Kingdom.
Barry
B
Fuller
Niteen Tapuria, Sameer Junnarkar, Mahmoud Abu-amara, Barry Fuller, Alexander M Seifalian, Brian R Davidson, Department of HPB Surgery and Liver transplantation, Royal Free Hospital, London NW32QG, United Kingdom.
Alexander M
AM
Seifalian
Niteen Tapuria, Sameer Junnarkar, Mahmoud Abu-amara, Barry Fuller, Alexander M Seifalian, Brian R Davidson, Department of HPB Surgery and Liver transplantation, Royal Free Hospital, London NW32QG, United Kingdom.
Brian R
BR
Davidson
D000818:Animals; D054360:Chemokine CXCL1 / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D006419:Heme Oxygenase (Decyclizing) / Q000378:metabolism*; D006439:Hemodynamics; D019194:Ischemic Preconditioning; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008102:Liver Circulation; D008107:Liver Diseases / Q000378:metabolism; D008297:Male; D008833:Microcirculation; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009504:Neutrophils / Q000378:metabolism*; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism
D054360:Chemokine CXCL1; C513297:Cxcl1 protein, rat; D016207:Cytokines; D006419:Heme Oxygenase (Decyclizing)
CINC; Haemoxygenase; Ischemic preconditioning; Microcirculation; Remote preconditioning; Reperfusion injury
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