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Effect of intraportal prostaglandin E1 administration on warm ischemic liver damage in the dog
Internatiord
Hepatology International Hepatology Communications 2 (1994) 257-262
ELSEVIER
communications
Effect of intraportal prostaglandin El administration warm ischemic liver damage in the dog
on
Eishi Totsuka*, Mutsuo Sasaki, Katsuro Takahashi, Yoshikazu Toyoki, Kageyoshi Seino, Shigeo Chiba, Syunji Narumi, Kenichi Hakamada, Mitsuru Konn The Depurtmeni
of Surgery,Hirosaki University School qj”Medicine, 5 Zaifu-cho. Hirosaki 036, Japan (Received 20 October 1993: accepted 7 January 1994)
Abstract To evaluate the effect of intraportal administration of prostaglandin E, (PGE,) on warm ischemic liver damage, two experimental studies were designed using dogs. First as a preliminary study (Expt. l), the portal blood flow and PGE, concentration in blood obtained from the portal vein and femoral artery were measured for each of portal and peripheral venous administration of PGE, at a rate of 0.02 ,ug/kg/ min. When PGE, was administered via the peripheral vein, little PGE, reached the liver, although portal blood flow was increased. Then, to determine the most effective route of PGE, administration to prevent 90-min warm ischemic liver damage, dogs were divided into the following three groups (Expt. 2): a PGE,-untreated (control) group (group A, n = lo), a peripheral venous PGE,-administered group (group B, n = 7) and an intraportal PGE,-administered group (group C, n = 7). PGE, was continuously infused before and after the ischemia at the rate of 0.02 @kg/min. The arterial ketone body ratio (acetoacetic acid//?-hydroxybutyric acid; AKBR) as well as the concentration of endotoxin (Etx) were measured. In both groups A and B, all the dogs died within 24 h. However, in group C, three out of the seven dogs survived and were sacrificed on the 3rd day. One dog died within 24 h, and the other three died within the next 2 days. The AKBR values were decreased after ischemia in all groups, but only in group C, this value recovered to the initial level. The Etx concentration increased in the early phase after ischemia in all groups, but in group C it started to decrease immediately after ischemia. The above results indicate that intraportal administration of PGE, provides a more protective effect than peripheral venous administration against warm ischemic liver injury. Key words: PGE,; Warm ischemic liver; Portal venous blood flow; Ketone body ratio; Endotoxin
*Corresponding author. 0928-4346/94/$07.00 0 1994 Elsevier Science B.V. All rights reserved SSDI 0928-4346(94)00008-S
258
E. Totsuka et al. lht.
Hepatol.
Commun. 2 (1994) 257-262
1. Introduction The protective effect of prostaglandin E, (PGE,) on liver damage has already been reported. however, when administered via a peripheral vein, about 70% of the PGE, is metabolized by the lung in the first pass immediately after injection [l]. This suggests that little PGE, actually reaches the liver. Therefore, as a preliminary study (Expt. 1) we investigated the pharmacokinetics of PGE, for each of portal and peripheral venous administration. Furthermore, to evaluate the direct effect of PGE, on hepatocytes, we attempted portal administration in the canine 90-min warm ischemic liver damage model (Expt. 2).
2. Materials and methods Adult mongrel dogs, each weighing about 10 kg, were used. Expt. 1
After laparotomy under general anesthesia, ileocecal vein was secured first as the portal PGEl administration route. The portal vein and femoral artery were also cannulated for each blood sampling. Both the portal blood flow and the concentration of PGE, in blood obtained from the portal vein and the femoral artery were measured during infusion of PGE, via the peripheral vein or via the portal vein at a rate of 0.02 &kg/mm. Expt. 2
After laparotomy under general anesthesia, the portal vein was cannulated first for PGE, administration, and then the femoral artery was cannulated for monitoring. As a model of warm ischemia, the hepaticoduodenal ligament was clamped to obstruct both the hepatic artery and portal vein for 90 min. Intestinal congestion at that time was evaded and drained through a portosystemic shunt, which had been created prior to clamping by connecting two silicone T-tubes, both inserted into the portal vein and inferior vena cava; this was removed just after declamping of the ligament. PGE, was infused for 30 min before clamping and for 12 h after declamping at a rate of 0.02 ~glkglmin. The dogs were divided into the following three groups; a PGE,-untreated (control) group (group A, n = lo), a peripheral PGE,-administered group (group B, n = 7) and an intraportal PGE,-administered group (group C, n = 7). The portal blood flow, the arterial ketone body ratio (acetoacetic acid@hydroxybutyric acid; AKBR) as well as the concentration of endotoxin (Etx) were measured successively before and after ischemia. The portal blood flow was measured using an electromagnetic blood flowmeter (MFV-3100, NIHON KODEN), PGE, using a [3H]prostaglandin E radioimmunoassay kit (Clinical Assays), AKBR by the modified Williamson method (KETO-340, IHARA ELECTRICAL) and Etx by Endospacy method. Values are expressed as means f SE. Student’s t test for unpaired data was used and P value of 5% or less was considered statistically significant.
E. Totsuka et al. lint. Hepatol. Commun. 2 (1994) 257-262
259
3. Results Expt. I
The portal blood flow increased only when PGE, was administered via the peripheral vein, but showed no change with portal infusion or in the control (Fig. 1, upper panel). The PGE, concentration in the portal vein did not increase, but that in the artery did increase, when PGE, was administered via the peripheral vein. When PGE, was administered via the portal vein, on the other hand, the arterial PGE, concentration showed no change (Fig. 1, lower panel). Expt. 2
In both groups A and B, all the dogs died within 24 h. However, in group C, three
(n=5) 1I
P co.05
0 0 A
P co.05
P co.
Artery Portal
51
001
P
I
Control PGE, i.v. PGE, i,p.
vein
co. 001 I 8420.0 k951.6
P < 0.05
P < 0.05
(meansfSEY)
Fig. 1. Changes in portal blood flow (upper panel) and PGE, concentration in both artery and portal vein (lower panel) when PGE, was infused via a peripheral vein or the portal vein at a rate of 0.02 pg/kg/min (n = 5). Control, no PGE, administration; PGE, i.v., intravenous PGE, administration; PGE, i.p., portal PGE, administration.
260
E. Totsuka et al. Ilnt. Hepatol. Commun. 2 (1994) 257-262 0%)
A(n=10) ---a---B (n-7) .,.,.,4,.,.,_ c (n-7) PCO.05 ** P-CO.005 l
a m ::
i
-’
.c
_c
(D
N
(meansfSEY)
Fig. 2. Changes in portal blood flow (upper panel), AKBR (middle panel) and Etx concentration (lower panel) in dogs with warm ischemic liver damage. A, no treatment (n = 10); B, intravenous PGE, administration (n = 7); C, portal PGE, administration (n = 7).
out of the seven dogs survived and were sacrificed on the 3rd day. One dog died within 24 h, and the other three died within the next 2 days. Mean survival time after ischemiawas6.4 k 3ShingroupA, 15.5 k 1.5hingroupBand42.8 k 9.2hingroup C. Portal blood flow started to increase immediately after declamping in all three groups. In group B, it was significantly high exactly 2 h after declamping compared with the others (Fig. 2, upper panel). The AKBR values showed different patterns in the three groups. First, they decreased in all the groups. In group A, this value was sustained at a low level even after declamping. In group B, it increased from the 1st hour, reached more than 1.O at the 2nd hour, and then decreased. In group C, on the other hand, the level continued to increase, and reached the initial level by the 3rd hour (Fig. 2, middle panel). The Etx concentration increased in the early phase after ischemia in all groups. In groups A and B, the values remained high. In group C,
E. Totsuka et al. IInt. Hepatol. Commun. 2 (1994) 257-262
261
however, the concentration started to decrease at the 2nd hour, and had dropped to the initial level by the 12th hour (Fig. 2, lower panel).
4. Discussion PGE, has both indirect and direct protective effects on hepatocytes. The former is an increase of portal blood flow [2], and the latter is stabilization of the hepatocellular membrane [3]. In our study, PGE, was infused at a rate of 0.02 ,@kg/min, because this dose was demonstrated to improve the microcirculation. Aida et al. reported that intravenous administration of PGE, increased portal blood flow, while intraportal administration did not [4]. The same results were obteined from Expt. 1 as to circulatory dynamics. On the other hand, it was supposed that PGE, reached the liver only from the hepatic artery after peripheral injection by measuring PGE, concentration in blood obtained from the portal vein and artery. Thus it is considered that the portal vein is the better route for PGE, administration to exert a direct protective effect on hepatocytes. Hypoxia and reperfusion injury associated with hepatic warm ischemia cause irreversible liver damage [5]. In our Expt. 2, all the dogs not given PGE, died in a short period after 90 min of warm liver ischemia, and the AKBR value did not increase at any time. As AKBR reflects oxidation-reduction ability [6], these data suggest that irreversible metabolic failure occurred in these dogs. When PGE, was administered via the peripheral vein, the AKBR value increased temporarily by the 2nd hour after declamping, and then decreased again. It is considered that the initial increase occurred as a result of the improved portal blood flow, whereas the decrease was caused by the reperfusion injury. When PGE, was administered via the portal route, on the other hand, the AKBR value started to increase immediately after ischemia, reaching the initial level, although the portal blood flow was the same as that in the control group. The liver detoxifies Etx originating from intestinal bacteria [7]. In our Expt. 2, the Etx concentration increased after ischemia. This was considered to be caused by Etx moving toward the systemic circulation from the portal vein through the bypass during ischemia. When PGE, was administered via the portal route, Etx decreased after ischemia. Here it was assumed that Etx detoxication in the liver was maintained by the portal administration of PGE,. In conclusion, the above results indicate that intraportal administration of PGE, provides a more protective effect than peripheral administration against warm ischemic liver damage, and could be adopted for protection of hepatocytes in fulminant hepatitis, hepatectomy with high risk, and liver transplantation.
References
[l] Golub M, Zia P,MatsunoM. 1975;56: 1404-1410.
Horton
R. Metabolism
of prostaglandin
A, and E, in man. J Clin Invest
262
E. Totsuka et al. Ilnt. Hepatol. Commun. 2 (1994) 257-262
[2] Arai Y, Asanuma Y, Nanjo H, et al. Experimental study on the effect of prostaglandin E, on decreased
hepatic tissue blood flow during operation. Jpn J Gastroenterol Surg 1992;25: 782-785. [3] Stachura J, Tamawski A, Kevin JI, et al. Prostaglandin protection of carbon tetrachloride-induced liver cell necrosis in the rat. Gastroenterology 1981;81:211-217. [4] Aida G, FA Bashour, BV Swamy, et al. Prostaglandin E,:Its effects on hepatic circulation in dogs. Pharmacology 1973;9: 336-347. [5] Nishida T, Kawashima Y, Tagawa K. Biochemical mechanism of ischemic cell injury. Nippon Taisha 1987;24: 379-387. [6] Kiuchi T, Ozawa K, Yamamoto Y, et al. Changes in arterial ketone body ratio in the phase immediately after hepatectomy. Arch Surg 1990;125:655-659. [7] Van Bossuyt H, Desmaretz C, Wisse E, et al. Response of cultured rat Kupffer cells to lipopolysaccharide. Arch Toxic01 1988: 62:316-324.
Hepatology International Hepatology Communications 2 (1994) 257-262
ELSEVIER
communications
Effect of intraportal prostaglandin El administration warm ischemic liver damage in the dog
on
Eishi Totsuka*, Mutsuo Sasaki, Katsuro Takahashi, Yoshikazu Toyoki, Kageyoshi Seino, Shigeo Chiba, Syunji Narumi, Kenichi Hakamada, Mitsuru Konn The Depurtmeni
of Surgery,Hirosaki University School qj”Medicine, 5 Zaifu-cho. Hirosaki 036, Japan (Received 20 October 1993: accepted 7 January 1994)
Abstract To evaluate the effect of intraportal administration of prostaglandin E, (PGE,) on warm ischemic liver damage, two experimental studies were designed using dogs. First as a preliminary study (Expt. l), the portal blood flow and PGE, concentration in blood obtained from the portal vein and femoral artery were measured for each of portal and peripheral venous administration of PGE, at a rate of 0.02 ,ug/kg/ min. When PGE, was administered via the peripheral vein, little PGE, reached the liver, although portal blood flow was increased. Then, to determine the most effective route of PGE, administration to prevent 90-min warm ischemic liver damage, dogs were divided into the following three groups (Expt. 2): a PGE,-untreated (control) group (group A, n = lo), a peripheral venous PGE,-administered group (group B, n = 7) and an intraportal PGE,-administered group (group C, n = 7). PGE, was continuously infused before and after the ischemia at the rate of 0.02 @kg/min. The arterial ketone body ratio (acetoacetic acid//?-hydroxybutyric acid; AKBR) as well as the concentration of endotoxin (Etx) were measured. In both groups A and B, all the dogs died within 24 h. However, in group C, three out of the seven dogs survived and were sacrificed on the 3rd day. One dog died within 24 h, and the other three died within the next 2 days. The AKBR values were decreased after ischemia in all groups, but only in group C, this value recovered to the initial level. The Etx concentration increased in the early phase after ischemia in all groups, but in group C it started to decrease immediately after ischemia. The above results indicate that intraportal administration of PGE, provides a more protective effect than peripheral venous administration against warm ischemic liver injury. Key words: PGE,; Warm ischemic liver; Portal venous blood flow; Ketone body ratio; Endotoxin
*Corresponding author. 0928-4346/94/$07.00 0 1994 Elsevier Science B.V. All rights reserved SSDI 0928-4346(94)00008-S
258
E. Totsuka et al. lht.
Hepatol.
Commun. 2 (1994) 257-262
1. Introduction The protective effect of prostaglandin E, (PGE,) on liver damage has already been reported. however, when administered via a peripheral vein, about 70% of the PGE, is metabolized by the lung in the first pass immediately after injection [l]. This suggests that little PGE, actually reaches the liver. Therefore, as a preliminary study (Expt. 1) we investigated the pharmacokinetics of PGE, for each of portal and peripheral venous administration. Furthermore, to evaluate the direct effect of PGE, on hepatocytes, we attempted portal administration in the canine 90-min warm ischemic liver damage model (Expt. 2).
2. Materials and methods Adult mongrel dogs, each weighing about 10 kg, were used. Expt. 1
After laparotomy under general anesthesia, ileocecal vein was secured first as the portal PGEl administration route. The portal vein and femoral artery were also cannulated for each blood sampling. Both the portal blood flow and the concentration of PGE, in blood obtained from the portal vein and the femoral artery were measured during infusion of PGE, via the peripheral vein or via the portal vein at a rate of 0.02 &kg/mm. Expt. 2
After laparotomy under general anesthesia, the portal vein was cannulated first for PGE, administration, and then the femoral artery was cannulated for monitoring. As a model of warm ischemia, the hepaticoduodenal ligament was clamped to obstruct both the hepatic artery and portal vein for 90 min. Intestinal congestion at that time was evaded and drained through a portosystemic shunt, which had been created prior to clamping by connecting two silicone T-tubes, both inserted into the portal vein and inferior vena cava; this was removed just after declamping of the ligament. PGE, was infused for 30 min before clamping and for 12 h after declamping at a rate of 0.02 ~glkglmin. The dogs were divided into the following three groups; a PGE,-untreated (control) group (group A, n = lo), a peripheral PGE,-administered group (group B, n = 7) and an intraportal PGE,-administered group (group C, n = 7). The portal blood flow, the arterial ketone body ratio (acetoacetic acid@hydroxybutyric acid; AKBR) as well as the concentration of endotoxin (Etx) were measured successively before and after ischemia. The portal blood flow was measured using an electromagnetic blood flowmeter (MFV-3100, NIHON KODEN), PGE, using a [3H]prostaglandin E radioimmunoassay kit (Clinical Assays), AKBR by the modified Williamson method (KETO-340, IHARA ELECTRICAL) and Etx by Endospacy method. Values are expressed as means f SE. Student’s t test for unpaired data was used and P value of 5% or less was considered statistically significant.
E. Totsuka et al. lint. Hepatol. Commun. 2 (1994) 257-262
259
3. Results Expt. I
The portal blood flow increased only when PGE, was administered via the peripheral vein, but showed no change with portal infusion or in the control (Fig. 1, upper panel). The PGE, concentration in the portal vein did not increase, but that in the artery did increase, when PGE, was administered via the peripheral vein. When PGE, was administered via the portal vein, on the other hand, the arterial PGE, concentration showed no change (Fig. 1, lower panel). Expt. 2
In both groups A and B, all the dogs died within 24 h. However, in group C, three
(n=5) 1I
P co.05
0 0 A
P co.05
P co.
Artery Portal
51
001
P
I
Control PGE, i.v. PGE, i,p.
vein
co. 001 I 8420.0 k951.6
P < 0.05
P < 0.05
(meansfSEY)
Fig. 1. Changes in portal blood flow (upper panel) and PGE, concentration in both artery and portal vein (lower panel) when PGE, was infused via a peripheral vein or the portal vein at a rate of 0.02 pg/kg/min (n = 5). Control, no PGE, administration; PGE, i.v., intravenous PGE, administration; PGE, i.p., portal PGE, administration.
260
E. Totsuka et al. Ilnt. Hepatol. Commun. 2 (1994) 257-262 0%)
A(n=10) ---a---B (n-7) .,.,.,4,.,.,_ c (n-7) PCO.05 ** P-CO.005 l
a m ::
i
-’
.c
_c
(D
N
(meansfSEY)
Fig. 2. Changes in portal blood flow (upper panel), AKBR (middle panel) and Etx concentration (lower panel) in dogs with warm ischemic liver damage. A, no treatment (n = 10); B, intravenous PGE, administration (n = 7); C, portal PGE, administration (n = 7).
out of the seven dogs survived and were sacrificed on the 3rd day. One dog died within 24 h, and the other three died within the next 2 days. Mean survival time after ischemiawas6.4 k 3ShingroupA, 15.5 k 1.5hingroupBand42.8 k 9.2hingroup C. Portal blood flow started to increase immediately after declamping in all three groups. In group B, it was significantly high exactly 2 h after declamping compared with the others (Fig. 2, upper panel). The AKBR values showed different patterns in the three groups. First, they decreased in all the groups. In group A, this value was sustained at a low level even after declamping. In group B, it increased from the 1st hour, reached more than 1.O at the 2nd hour, and then decreased. In group C, on the other hand, the level continued to increase, and reached the initial level by the 3rd hour (Fig. 2, middle panel). The Etx concentration increased in the early phase after ischemia in all groups. In groups A and B, the values remained high. In group C,
E. Totsuka et al. IInt. Hepatol. Commun. 2 (1994) 257-262
261
however, the concentration started to decrease at the 2nd hour, and had dropped to the initial level by the 12th hour (Fig. 2, lower panel).
4. Discussion PGE, has both indirect and direct protective effects on hepatocytes. The former is an increase of portal blood flow [2], and the latter is stabilization of the hepatocellular membrane [3]. In our study, PGE, was infused at a rate of 0.02 ,@kg/min, because this dose was demonstrated to improve the microcirculation. Aida et al. reported that intravenous administration of PGE, increased portal blood flow, while intraportal administration did not [4]. The same results were obteined from Expt. 1 as to circulatory dynamics. On the other hand, it was supposed that PGE, reached the liver only from the hepatic artery after peripheral injection by measuring PGE, concentration in blood obtained from the portal vein and artery. Thus it is considered that the portal vein is the better route for PGE, administration to exert a direct protective effect on hepatocytes. Hypoxia and reperfusion injury associated with hepatic warm ischemia cause irreversible liver damage [5]. In our Expt. 2, all the dogs not given PGE, died in a short period after 90 min of warm liver ischemia, and the AKBR value did not increase at any time. As AKBR reflects oxidation-reduction ability [6], these data suggest that irreversible metabolic failure occurred in these dogs. When PGE, was administered via the peripheral vein, the AKBR value increased temporarily by the 2nd hour after declamping, and then decreased again. It is considered that the initial increase occurred as a result of the improved portal blood flow, whereas the decrease was caused by the reperfusion injury. When PGE, was administered via the portal route, on the other hand, the AKBR value started to increase immediately after ischemia, reaching the initial level, although the portal blood flow was the same as that in the control group. The liver detoxifies Etx originating from intestinal bacteria [7]. In our Expt. 2, the Etx concentration increased after ischemia. This was considered to be caused by Etx moving toward the systemic circulation from the portal vein through the bypass during ischemia. When PGE, was administered via the portal route, Etx decreased after ischemia. Here it was assumed that Etx detoxication in the liver was maintained by the portal administration of PGE,. In conclusion, the above results indicate that intraportal administration of PGE, provides a more protective effect than peripheral administration against warm ischemic liver damage, and could be adopted for protection of hepatocytes in fulminant hepatitis, hepatectomy with high risk, and liver transplantation.
References
[l] Golub M, Zia P,MatsunoM. 1975;56: 1404-1410.
Horton
R. Metabolism
of prostaglandin
A, and E, in man. J Clin Invest
262
E. Totsuka et al. Ilnt. Hepatol. Commun. 2 (1994) 257-262
[2] Arai Y, Asanuma Y, Nanjo H, et al. Experimental study on the effect of prostaglandin E, on decreased
hepatic tissue blood flow during operation. Jpn J Gastroenterol Surg 1992;25: 782-785. [3] Stachura J, Tamawski A, Kevin JI, et al. Prostaglandin protection of carbon tetrachloride-induced liver cell necrosis in the rat. Gastroenterology 1981;81:211-217. [4] Aida G, FA Bashour, BV Swamy, et al. Prostaglandin E,:Its effects on hepatic circulation in dogs. Pharmacology 1973;9: 336-347. [5] Nishida T, Kawashima Y, Tagawa K. Biochemical mechanism of ischemic cell injury. Nippon Taisha 1987;24: 379-387. [6] Kiuchi T, Ozawa K, Yamamoto Y, et al. Changes in arterial ketone body ratio in the phase immediately after hepatectomy. Arch Surg 1990;125:655-659. [7] Van Bossuyt H, Desmaretz C, Wisse E, et al. Response of cultured rat Kupffer cells to lipopolysaccharide. Arch Toxic01 1988: 62:316-324.