Stage-classification of reticuloendothelial function in hepatic injuries in the rat

The serial changes of RE function were classified into 3 stages according to changes in opsonic activity and phagocytic index in hepatectomized, CCl4-treated cirrhotic rats; enhanced stage, compensating stage and critical stage. The phagocytic index was measured by initial disappearance rate after 4 mg/kg i.v. injection of 51Cr-labelled LPS. An opsonic activity was determined by the bioassay method using cultured RE cells prepared from the normal rat liver. The enhanced stage was characterized by an enhancement in both phagocytic index and opsonic activity--3 to 21 day-70% hepatectomized, 2 to 5 day-40% hepatectomized, 3 week-cirrhotic rats. The compensating stage by increased opsonic activity and normal phagocytic index was 2 day-70% hepatectomized and 9 week-cirrhotic rats. The critical stage by decreased phagocytic index was 5 hour to 1 day-70% hepatectomized, 13 week-cirrhotic rats.     

Effect of hepatectomy on the reticuloendothelial system of septic rats

The purpose of this study was to evaluate the function of the reticuloendothelial system (RES) of sham hepatectomy and 20% and 50% partial hepatectomy (PH 20%, PH 50%), with or without cecal ligation and puncture-induced sepsis. The animals were injected with 51Chromium sheep red blood cells (SRBC) at 72 hours. SRBC half life (T1/2) was measured as an index of RES function and the percentage distribution of SRBC in liver, lung, and spleen was calculated. T1/2 was significantly prolonged in PH 50% rats and was associated with decreased radioactive uptake by the liver. Mortality was nil in the control groups and markedly increased in the presence of sepsis. The results suggest that decreased RES function following hepatectomy is dependent upon the proportion of liver removed and that sepsis further increased the mortality of hepatectomized animals.     

Gadolinium chloride prevents mortality in hepatectomized rats given endotoxin

The purpose of this study was to determine if inhibition of Kupffer cells by gadolinium chloride (GdCl(3)) affects the arterial ketone body ratio (AKBR), liver injury, and mortality in hepatectomized rats administered lipopolysaccharide (LPS). Rats treated with or without GdCl(3) received a 70% hepatectomy. Either LPS (5 mg/kg) or vehicle (saline) was administered 48 h after hepatectomy. Further, hepatectomized rats were administered superoxide dismutase (CuZnSOD, 9 x 10(4) U/kg) before and every 3 h after LPS injection up to 9 h to assess involvement of superoxide in liver injury in this model. All hepatectomized rats with saline died within 24 h after LPS administration. In contrast, GdCl(3) prevented this mortality completely. Serum AST levels were about 160 IU/L in hepatectomized rats with vehicle; however, values were increased approximately 25-fold by LPS administration. In contrast, these increases were blunted significantly by about 90% by GdCl(3). Further, GdCl(3) also prevented decreases in AKBR caused by LPS. LPS caused severe liver injury, which was stopped almost completely by GdCl(3). LPS-induced increases in superoxide production by isolated Kupffer cells were stopped by about 90% by GdCl(3). Importantly, SOD administration prevented decreases in AKBR, liver injury, and mortality significantly as well as GdCl(3). These results indicated that GdCl(3) prevented liver injury and mortality caused by LPS most likely by inhibiting superoxide production by Kupffer cells. Thus, inhibition of activation of Kupffer cells could be useful for preventing liver dysfunction in postoperative endotoxemia.     

Experimental and clinical studies on the effect of reticuloendothelial system (RES) potentiator in the depressed RES function in cirrhotics

Among the patients with liver cirrhosis (LC) who undergo the operation, the postoperative complications are not infrequent and sometimes prove fatal. The impaired hepatic function, especially the impaired reticuloendothelial system (RES) function, has been claimed to be a possible pathogenic factor for these complications. The present experimental and clinical studies were undertaken to investigate the RES function and the effect of preoperative OK-432 administration as an RES potentiator in LC. The results are as follows: 1) CCl4-induced LC rats were evaluated for RES global phagocytic function, Kupffer cell phagocytic function, plasma opsonic activity and plasma opsonic substances such as fibronectin, C3 and IgG. All parameters except IgG showed significant depression compared to those values in normal rats. However, the administration of OK-432 (0.1 KE/rat, ip) improved all these depressed parameters. The OK-432 administration also significantly improved the survival following panperitonitis in LC rats. 2) Among 18 LC patients with hepatocellular carcinoma undergoing partial hepatectomy, the RES global phagocytic function, plasma opsonic activity and plasma opsonic substances were evaluated. Same as the experimental study, all parameters except IgG were significantly depressed among the LC patients compared to those values in the patients with normal liver. However, the preoperative OK-432 administration (5 KE/day sc for 4 days) significantly improved these parameters and consequently decreased the postoperative complications. These results indicate that the preoperative RES activation by the OK-432 was effective and useful for the prevention of the postoperative complications in the LC patients.     

The three different phases of reticuloendothelial system phagocytic function in rats with liver injury

In the present study, reticuloendothelial system (RES) phagocytic function of rats with partial hepatectomy or experimentally induced liver cirrhosis was investigated by determining the phagocytic index, the opsonic index, and uptake rate in liver, spleen, and lung of a 51Cr-labeled endotoxin-injected rat. In both the partially hepatectomized and the cirrhotic rats, all three indicators varied markedly according to the elapsed period since liver injury. The changes in RES phagocytic function were classified into three different phases: compromised, compensatory, and enhanced. The compromised phase, consisting of a decrease in the phagocytic index, was observed during the first 24 hr after 67% hepatectomy and in advanced liver cirrhosis. This represented the failure of RES phagocytic function. The compensatory phase, in which the phagocytic index was maintained at nearly normal levels mainly by a compensatory enhancement in the opsonic index, was seen during the first to second postoperative day and in moderate liver cirrhosis. The enhanced phase, with a high phagocytic index, was observed from Day 4 to approximately Day 14 after surgery, and in the cases of mild liver damage. In the compromised and compensatory phases, the liver uptake rate was significantly decreased compared with the control. However, the uptake in the spleen and lung were markedly increased. In conclusion, the phagocytic function of the RES was significantly affected to a degree which changed with the extent of liver damage.     

Effect of administration of fibronectin or aprotinin on liver regeneration after experimental hepatectomy.

Following 70% hepatectomy, male Sprague-Dawley strain rats were given purified human plasma fibronectin and/or aprotinin intraperitoneally after hepatectomy and were divided into 4 groups: Group FA (40 mg/kg of FN and 25,000 KIU/kg of aprotinin), Group F (40 mg/kg of fibronectin), Group A (25,000 KIU/kg of aprotinin), and Group C which served as a control. The liver regeneration rate 72 hours after hepatectomy in Groups FA and F, and the plasma FN levels at 24 and 72 hours following hepatectomy in groups with FN, were significantly higher than that of the control group. The incorporation of 3H-thymidine into DNA and the phagocytic index of the reticuloendothelial system at 24 hours in Groups FA and F, and the mitotic index at 72 hours after hepatectomy in Group FA were significantly increased as compared to the control. But the supplementation of 20 mg/kg of indomethacin to the 4 groups resulted in significant suppression of 3H-thymidine uptake 24 hours after hepatectomy. Moreover, plasma fibronectin levels in rats correlated well with the phagocytic index and DNA synthesis 24 hours following partial hepatectomy. These results suggest that fibronectin may act as an activator of the reticuloendothelial system, related to prostaglandins, in the process of liver regeneration.     

Reticuloendothelial Clearance of Blood-borne Particulates: Relevance to Experimental Lung Microembolization and Vascular Injury

The rapid increase in sheep lung vascular permeability observed during Pseudomonas aeruginosa bacteremia may be due to embolization of the pulmonary microvasculature by bloodborne particulates. Since alterations in lung microvascular permeability during mild septicemia in sheep may reflect inefficient RES phagocytic clearance of bacteria as well as products of bacterial induced intravascular coagulation, the opsonic and phagocytic aspects of RES function in sheep (30-50 kg) were compared to other species. RES function was evaluated by both the clearance and relative organ uptake of gelatinized I¹³¹ RE test lipid emulsion and gelatinized colloidal carbon. Immunoreactive opsonic a₂SB glycoprotein levels were determined by electroimmunoassay. The phagocytic index for RES clearance of the gelatinized (500 mg/kg) test lipid in sheep was 0.019 ± 0.002 corresponding to a half-time of 16.65 ± 1.74 minutes. With colloidal carbon (64 mg/kg), the phagocytic index in sheep was 0.080 ± 0.026, corresponding to a half-time of 6.16 ± 1.99 minutes. The per cent of injected lipid emulsion (%ID) in major RE organs, on a total organ basis (TO), was: liver = 15.69 ± 1.65%; spleen = 2.09 ± 0.78%. Localization in the lung = 31.39 ± 6.2%. The per cent of carbon localized in major RE organs (%ID/TO) was: liver = 21.37 ± 1.9%; spleen = 1.95 ± 0.55%. Localization in the lung = 32.70 ± 4.55%. In contrast, clearance and organ distribution of the blood-borne test microparticles in rats and dogs at the same relative challenging dose revealed a much more intense and rapid liver and spleen RES uptake with minimal lung localization (1-2%). Immunoreactive opsonic protein concentrations varied greatly with species and directly correlated with efficiency of RES function. Levels observed were: dog = 1285 ± 135 µg/ml; mouse = 1077 ± 67 µg/ml; rat = 400 ± 31 µg/ml; human = 297 ± 10 µg/ml; and sheep = 184 ± 13 µg/ml. After intravenous particulate challenge, circulating immunoreactive opsonic protein in the sheep was depleted (p < 0.05) rapidly with partial recovery at 24 hours and mild rebound hyperopsonemia at 48 hours. This pattern is in contrast to the rapid restoration seen in dog and rat within three to six hours postchallenge. Thus, in sheep, the extensive pulmonary localization of blood-borne microparticles appears related to inefficient RES clearance function mediated by a relative deficiency of circulating opsonic protein (plasma fibronectin).     

Treatment of post-hepatectomy hepatic cellular energy crisis in cirrhosis with ATP-MgCl2 administration

The present study was undertaken to investigate the effect of ATP-MgCl2 for hepatic cellular energy crisis following hepatectomy in cirrhosis. In experimental study, cirrhotic rats, induced by subcutaneous injection of CCl4 twice a week for 10 weeks, received ATP-MgCl2 (12.4 mumoles) (ATP group) or saline (control group) at 2 hours after 68% hepatectomy. The hepatic cellular energy charge (EC) and arterial ketone body ratio (AKBR), which were good indicators to evaluate the hepatic cellular metabolism, and reticuloendothelial system (RES) function were significantly improved among ATP group at 24 hours after hepatectomy compared to the control group. Survival at one week was also significantly improved with ATP-MgCl2 treatment. In clinical study, ATP-MgCl2 (30-50 mumoles/kg) was infused intravenously to twenty hepatectomized cirrhotic patients. The hepatic cellular energy metabolism, studied using AKBR, and RES function as well as the clinical course were improved with ATP-MgCl2) treatment compared to those of the conventionally treated controls. These data indicate that ATP-MgCl2 is beneficial as one of the therapeutic approaches to improve the hepatic cellular energy crisis after hepatectomy which is the major cause of death following hepatectomy among cirrhotic patients.     

32P-玻璃微球间质给药体内生物学分布与代谢

目的 观察32P-玻璃微球(32P-GMS)间质给药后体内生物学分布与代谢.方法 120只小鼠分别经肝脏或肌肉注射32P-GMS,每只(1.80±0.05)MBq/50μl,不同时间点处死取血及脏器计算每克组织百分注射剂量率(%ID/g);12只大鼠肝脏、肌肉每只注射(18.0±0.5)MBq/0.5 ml,收集排泄物测累积排泄率.结果 小鼠肝脏组给药肝叶%ID/g 0时最高为1.38,15 nin降至0.71,非给药肝叶峰值15 min为0.52,4 h之后左右肝叶无区别;肺组织计数率值1 h内上升,肝肺累积分流率为37.9%.肌肉组注射点%ID/g0时为31.47,15 nin～8 h为25.06～11.92(中值20.97),12 h为8.70,24 h～14 d为3.54～2.02(中值2.51),其他主要脏器放射性接近本底水平.大鼠肝脏组粪便和尿液14 d累积排泄率分别为0.0751%和0.0586%;肌肉组分别为0.0401%和0.0385%.结论 32P-GMS间质注射适用于除肝脏以外的体内组织脏器恶性实体瘤的治疗.     

Enhancement of portal blood flow by ursodesoxycholic acid in partially hepatectomized rats

Portal venous flow (PVF) was examined after portal injection of ursodesoxycholic acid (URSO) in rats that were partially hepatectomized by either 40% or 66%. URSO (10 mg/kg per minute) was injected into the portal vein and was thereafter observed to increase PVF concomitantly with a fall in portal venous pressure (PVP) in control animals. The increase in PVF in response to URSO was dose-dependent. In hepatectomized rats, the PVF response was augmented when the same dose of URSO was portally injected, and the magnitude of response was enhanced in proportion to the volume of liver resected. These results suggest that URSO increases PVF through vasodilation of the portal vessels, and therefore URSO is considered to increase PVF potently in a partially hepatectomized condition.     

Regenerative capacities of normal and cirrhotic livers following 70% hepatectomy in rats and the effect of alpha-tocopherol on cirrhotic regeneration

BACKGROUND: The regeneration of normal and cirrhotic liver has been very well demonstrated after partial hepatectomy; although the tissue regenerated by cirrhotic liver is also cirrhotic. The structural differences of the regenerated tissues between normal and cirrhotic livers may also indicate different regeneration capacities. The objective of this study was to compare the regeneration capacities of normal and cirrhotic livers by bromodeoxyuridine (BrdU) incorporation and proliferating cell nuclear antigen (PCNA) labeling indices in replicating nuclei and mitotic figures in cells in partially hepatectomized normal and cirrhotic rats and to study the effect of alpha-tocopherol on cirrhotic liver regeneration. METHODS: Five groups of adult Wistar rats comprised normal livers, cirrhotic livers, regenerated normal livers, regenerated cirrhotic livers, and alpha-tocopherol-treated regenerated cirrhotic livers. Cirrhosis was induced by intragastric administration of carbon tetrachloride and phenobarbital in the drinking water of the rats. Liver regeneration capacities in normal and cirrhotic rats and following partial hepatectomy in normal and cirrhotic rats and cirrhotic rats that were administered alpha-tocopherol were evaluated through BrdU incorporation, PCNA labeling, and mitotic indices. RESULTS: BrdU and PCNA labeling and mitotic indices were zero for normal rats and 4.3 +/- 3.5, 6.5 +/- 5, and 2.5 +/- 1.5 for cirrhotic rats, respectively. The values after partial hepatectomy in normal and cirrhotic rats were 46.2 +/- 8.7 and 27.8 +/- 7.5 for BrdU labeling, 83.7 +/- 6.5 and 51.3 +/- 6.8 for PCNA labeling, and 31.8 +/- 4.2 and 18.6 +/- 3.4 for mitotic index, respectively. For the fifth group comprising cirrhotic rats that were administered alpha-tocopherol and had undergone partial hepatectomy, BrdU incorporation, PCNA labeling, and mitotic indices were 37.5 +/- 6.3, 76.5 +/- 6.2, and 27.2 +/- 4.2, respectively. When the cirrhotic liver regeneration group was compared with the normal liver regeneration group, rates of liver regeneration in the cirrhotic group were significantly depressed (P < 0.01). Although the BrdU incorporation and PCNA labeling indices of the alpha-tocopherol-administered cirrhotic liver regeneration group indicated significantly lower rates of liver regeneration when compared with the normal liver regeneration group (P < 0.05), the liver regeneration rates of the alpha-tocopherol-administered cirrhotic group were also significantly higher than those of the cirrhotic liver regeneration group that was not administered alpha-tocopherol (P < 0.01). CONCLUSIONS: Cirrhotic livers revealed a significantly depressed capacity for regeneration following partial hepatectomy. alpha-Tocopherol administration seemed to improve the rates of regeneration in cirrhotic rats with respect to the BrdU incorporation, PCNA labeling, and mitotic indices.     

High expression of the CD14 gene and interleukin-1beta gene in the liver and lungs of cirrhotic rats after partial hepatectomy

BACKGROUND: A hepatic resection is invasive for cirrhotic patients because postoperative complications, such as hepatic disturbance sometimes resulting in hepatic failure and pulmonary disturbances, are frequent and serious. We investigated here the alteration of the CD14 and inflammatory cytokine genes expressed in the liver and lungs after partial hepatectomy (PH) of a cirrhotic rat model to help elucidate the pathophysiological change occurring during the postoperative course of hepatectomized cirrhotic patients. MATERIALS AND METHODS: Wistar rats were orally administrated carbon tetrachloride once a week for 14 weeks to induce liver cirrhosis. In comparison with cirrhotic and normal rats, we analyzed the expression of the CD14, tumor necrosis factor-alpha, and interleukin (IL)-1beta genes in remnant liver and whole lung tissue during 48 h after 30% partial hepatectomy with Northern blottings and measured asparatate aminotransferase (AST) in serum for evaluation of postoperative hepatic injury. Gadolinium chloride (GdCl3; 7 mg/kg body weight) was intravenously injected 24 h before partial hepatectomy to suppress Kupffer cells (KC) activation. RESULTS: The expression of the CD14 and IL-1beta genes moderately increased at 6 h and peaked at 12 h in parallel with the time course of AST values after PH only in cirrhotic rats. GdCl(3) significantly inhibited the elevation of AST similar to the inhibition of the expression of the CD14 and IL-1beta genes after PH. In addition, the expression of these genes showed marked enhancement in the lungs of the cirrhotic hepatectomy model. CONCLUSIONS: KC activation was responsible for hepatic injury after PH, and the CD14 system appeared to be an early trigger for KC activation followed by induction of inflammatory cytokine IL-1beta synthesis leading to hepatic injury. Furthermore, the CD14 system was suggested to participate in respiratory disturbances after hepatectomy in cirrhosis.     

Ischemic injury in cirrhotic livers: an experimental study of the temporary arrest of hepatic circulation.

In order to prevent massive bleeding at hepatectomy, the temporary arrest of hepatic circulation is often performed but it is not clear whether this arrest of circulation is tolerated equally by the cirrhotic liver and the normal liver. We investigated biochemically the detailed effects of ischemia on cirrhotic livers in rats with experimentally induced liver cirrhosis. Thioacetoamide was used to prepare the rat cirrhotic liver model (LC group, n = 6). After laparotomy, the vessels to the left lateral lobe were clamped for 30 min and then declamped. Changes in AST isozymes and the aminogram in the blood were examined after ischemia. Postischemic changes in hepatic adenine nucleotides (AdN) and the brain aminogram were also examined. These were compared with those of normal liver ischemia (N group, n = 6) at 24 hr after recirculation. In the LC group, serum levels of mitochondrial AST, a parameter of necrotic cells, were significantly higher than those of the N group. Hepatic AdN levels decreased to 60.6% of the original levels after ischemic injury but those of the N group remained at 95.4% of the original level. Since AdN in tissue is accepted as a reliable parameter of viable cells, cirrhotic livers subjected to ischemia might have more necrotic cells than normal livers. Sequential analysis of serum aminograms of the LC group after ischemia revealed that the ratio of Val+Leu+Ileu/Tyr+Phe decreased to near 1.0 but that of the N group always remained higher than 3.0. Based on these results, it was concluded that ischemic injury in cirrhotic livers is more hazardous than that in normal livers.     

Bacterial clearance in the intact and regenerating liver

The Kupffer cells in the liver play an important role in reticuloendothelial system (RES) function by clearing particulate matter and bacteria from the blood stream. While hepatocyte regeneration and function have been extensively studied following partial hepatectomy, little information is available concerning RES function in the regenerating liver. This study investigates hepatic RES function by evaluating bacterial clearance (live E. coli) in the intact and regenerating liver. Thirty-four young male Sprague Dawley rats were studied. Twenty-two animals underwent a standard 70% partial hepatectomy using ligature technique and 12 had a sham operation. Both groups of rats received 10(9) organism of S35 labeled E coli, intravenously at 24 hours, 72 hours, 2 1/2 weeks, and 6 weeks postoperatively. Rats were killed 10 minutes following injection and liver, lung, spleen, and kidney harvested, fixed, and radioactivity was determined using a scintillation spectrometer interfaced with a micro-computer counting the S35 radiolabel. The total organ count of trapped bacteria in liver in partially hepatectomized rats was lower than intact controls at 24 hours (22.0% v 46.4%, P less than .01), but was similar at 72 hours, 2 1/2 weeks, and 6 weeks. Partial hepatectomy increased the amount of bacterial trapping in the lung at 24 hours (11.3% v 1.7%, P less than .01) and 72 hours (10.1% v 1.7%, P less than .05) and returned to normal at 2 1/2 weeks and 6 weeks. Splenic activity was increased following hepatectomy at 2 1/2 weeks. Renal clearance was increased at 72 hours and 2 1/2 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)     

Analysis of factors relating to early repairment of the surgical margin following liver resection--factors relating to duration of hospital stay

To analyze relating factors to early repairment of the surgical margin of the remnant liver we measured plasma fibronectin (FN), coagulation factor XIII (XIII), polymorphonuclear leukocyte elastase (PMNE), platelet counts (Plt), prothrombin time (PT%) before and at the first, third, 7th and 14th days after liver resection in 25 patients. Changes in these factors (1) were compared with their clinical status, such as liver cirrhosis, high fever, abscess formation, duration of drainage and use of microwave tissue coagulator (MTC) (2). The multivariate analysis about the factors influencing the duration of hospital stay (3) were carried out. (1) In the all cases FN, XIII (14th), PT%, Plt decreased and PMNE increased significantly versus pre-operative data. There were significant correlations between FN and XIII, FN and PMNE. (2) In the cirrhotic group FN, XIII, PT% and Plt were significantly lower than those of non cirrhotic group. In the abscess formation group PT% was significantly lower than the no abscess formation group. In the MTC group XIII, PT% and Plt were significantly lower than those of the no-MTC group. In conclusion, FN, XIII, Plt and PT% in the cirrhotic, high fever, abscess formation or longer drainage group were in lower levels compared with the each control group. (3) By the multivariate analyses, abscess formation, high fever and liver cirrhosis were the most influencing factors for the duration of hospital stay.     

极化液对内毒素血症大鼠肝损伤的影响

目的研究葡萄糖-胰岛素-钾(极化液,GIK)对内毒素血症大鼠肝损伤的影响。方法雄性SD大鼠60只,体重200~250g,随机分为三组:对照组,脂多糖组(LPS组,LPS 8mg/kg),GIK组(LPS 8mg/kg+GIK 4ml·kg~(-1)·h~(-1))。采用全自动生化仪检测腹腔注射LPS后3d和5d大鼠血清丙氨酸氨基转移酶(ALT)及天门冬氨酸氨基转移酶(AST)含量,ELISA法检测三组大鼠肝组织匀浆TNF-α含量,并行HE染色观察肝组织病理变化,TUNEL免疫荧光检测肝实质细胞凋亡情况。结果与注射后3d比较,注射后5dLPS组大鼠血清ALT、AST、肝组织匀浆TNF-α含量明显升高,而GIK组大鼠血清ALT、AST、肝组织匀浆TNF-α含量明显下降(P0.05)。与对照组比较,注射后3dLPS组和GIK组大鼠血清ALT、AST、注射后5dLPS组大鼠血清ALT、AST明显升高(P0.05),注射后3、5dLPS组和GIK组大鼠肝组织匀浆TNF-α含量、肝损伤等级评分、肝细胞凋亡指数明显升高(P0.05)。与LPS组比较,注射后3dGIK组大鼠肝组织匀浆TNF-α含量、肝细胞凋亡指数明显降低(P0.05),注射后5dGIK组大鼠血清ALT、AST、肝组织匀浆TNF-α含量、肝损伤等级评分、肝细胞凋亡指数明显降低(P0.05)。结论腹腔注射LPS可引起大鼠肝损伤,导致肝功能改变及肝细胞破坏;GIK可减轻LPS诱导的大鼠肝损伤。     

Changes of gluconeogenesis and alanine metabolism following partial hepatectomy in normal and cirrhotic rats

Gluconeogenesis and alanine metabolism of normal and cirrhotic rats were studied in view of partial hepatectomy. Liver cirrhosis was made by repeated injection of thioacetamide in rat. Partial hepatectomy was performed by modified method of Higgins-Anderson. Liver glycogen and fructose-2, 6-bisphosphate were decreased after hepatectomy and recovered within 7 days in normal groups, while those of cirrhotic group reduced even in preoperative state were further decreased and hardly recovered after hepatectomy. Gluconeogenesis of perfused liver in cirrhosis was increased from both lactate and alanine preoperatively, but gluconeogenesis from alanine was not increased in both hepatectomized rats. ATP and energy charge were decreased after hepatectomy and recovered within two weeks. These level were lower in cirrhotic group, and decreased further and hardly recovered after hepatectomy. Alanine utilization to CO2 in vivo was not impaired in cirrhotic group either preoperatively or postoperatively. ATP and energy charge were increased by alanine injection in hepatectomized rats of both normal and cirrhotic group. In conclusion, glucose-insulin therapy of sufficient amounts is important to improve decreased glycolysis and abnormal gluconeogenesis on both post-hepatectomy period of normal and pre and post-hepatectomy period of cirrhosis. Also alanine is effective for stimulating decreased energy production.     

Protection of sinusoidal endothelial cells against storage/reperfusion injury by prostaglandin E2 derived from Kupffer cells.

BACKGROUND: In clinical liver transplants, grafts are frequently exposed to endotoxin (lipopolysaccharide, LPS) before harvest and may be predisposed to dysfunction. Because graft failure is linked to sinusoidal endothelial cell injury after storage/reperfusion, we investigated the effect of donor exposure to LPS on graft survival in relation to sinusoidal endothelial cell injury after storage/reperfusion in rats. METHODS: Rats were injected with 0.5 mg/kg LPS. In some rats, 20 mg/kg GdCl3 or 5 mg/kg indomethacin was injected before LPS to ablate Kupffer cells and inhibit prostaglandin (PG) synthesis, respectively. Other rats were injected with 100 microg/kg dimethyl PGE2, a stable PGE2 analog. Rat livers were harvested, stored in cold UW solution and transplanted to non-treated rats for determination of survival and liver injury in recipients. Otherwise, after cold storage, the livers were reperfused briefly with physiological buffer containing trypan blue for determination of sinusoidal endothelial cell injury by counting trypan blue-positive nuclei in histological sections. RESULTS: Donor treatment with LPS increased hepatic PGE2 production before storage and decreased recipient survival, but paradoxically decreased killing of sinusoidal endothelial cells after storage and reperfusion. Pretreatment of donors with GdCl3 or indomethacin prevented the protective preconditioning of sinusoidal endothelial cells by LPS, whereas pretreatment with dimethyl PGE2 protected sinusoidal endothelial cells to the same extent as LPS. Unlike LPS, however, PGE2 attenuated graft injury after liver transplants. CONCLUSION: PGE2 derived from LPS-stimulated Kupffer cells protects sinusoidal endothelial cells against storage/reperfusion injury. Unlike LPS, PGE2 improves graft function after liver transplants. Thus, donor preconditioning with PGE2 may be beneficial in liver transplants.     

烟碱对内毒素血症大鼠凝血功能的影响

目的 探讨烟碱对内毒素血症大鼠凝血功能的影响.方法 成年健康雄性SD大鼠96只,体重200～250 g,采用随机数字表法,将大鼠随机分为4组(n=24):生理盐水对照组(NS组)、内毒素组(LPS组)、烟碱组(NIC组)、α7烟碱型乙酰胆碱受体(α7nAchR)拮抗剂组(α-BGT组).LPS组、NIC 组和α-BGT组经股静脉注射脂多糖(LPS)10 mg/kg制备内毒素血症模型.α-BGT组于注射LPS前45min腹腔注射α-BGT 1μg/kg,注射α-BGT后15 min腹腔注射烟碱400 μg/kg.NIC组除用等容量生理盐水替代α-BGT外,余操作同α-BGT组.LPS组除用等容量生理盐水替代烟碱外,余操作同NIC组.NS组除用等容量生理盐水替代LPS外,余操作同LPS组.于注射LPS前、注射后2、4、6 h时采集腹主动脉血样,检测血浆凝血酶原时间(PT)、部分活化凝血活酶时间(APTT)、纤维蛋白原(Fib)、抗凝血酶(AT)、血管性血友病因子(vWF)、纤溶酶原激活物抑制物-1(PAI-1)、D-二聚体、肿瘤坏死因子-α(TNF-α)水平和血小板计数.结果 与NS组比较,LPS组和α-BGT组PT、APTT延长,血浆Fib、AT水平降低,血小板计数减少,血浆PAI-1、WF、D-二聚体与TNF-α水平升高(P＜0.05).与LPS组比较,NIC组PT、APTT缩短,血浆Fib和AT水平升高,血小板计数增加,血浆PAI-1、vWF、D-二聚体和TNF-α水平降低(P＜0.05).与NIC组比较,α-BGT组PT和APTT延长,血浆Fib和AT水平降低,血小板计数减少,血浆PAI-1、vWF、D-二聚体和TNF-α水平升高(P＜0.05).结论 烟碱可抑制内毒素血症大鼠过度激活的凝血反应,缓解抗凝血与纤维蛋白溶解功能的抑制状态,作用机制可能与其结合外周α7烟碱型乙酰胆碱受体激活胆碱能抗炎通路,从而减少促炎性细胞因子释放有关.

Abstract：

Objective To investigate the effect of nicotine on coagulation abnormalities in endotoxemic rats.Methods Ninety-six male SD rats weighing 200-250 g were randomly divided into 4 groups (n=24 each): group normal saline (group NS);group LPS;group nicotine(group NIC)and group α-bungarotoxin (α7 nicotinic acetylcholine receptor antagonist, group α-BGT) . Endotoxemia was induced by LPS 10 mg/kg injected via femoral vein in LPS, NIC and α-BGT groups. In group NIC nicotine 400 μg/kg was injected intraperitoneally at 30 min before LPS injection. In group α-BGT α-BGT 1 μg/kg was injected intraperitoneally at 15 min before intraperitoneal nicotine. Prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(Fib),antithrombin (AT),von Willebrand factor(vWF),plasminogen activator inhibitor-1(PAI-1),D-dimer,platelet count and TNF-α were measured before (baseline) and 2, 4 and 6 h after LPS injection.Results PT and APTT were significantly prolonged and plasma Fib and AT concentrations and platelet count were significantly decreased, while plasma PAI-1, D-dimer, vWF and TNF-α concentrations were significantly increased after LPS administration in group LPS as compared with group NS. Nitotine pretreatment significantly attenuated the LPS-induced changes in group NIC.The effect of nicotine was counteracted by α-BGT. Conclusion Nicotine can attenuate coagulation abnormalities induced by LPS by acting on α7 nicotinic acetylcholine receptor.