Effect of Acute Ethanol Exposure on Hepatic Stimulator Substance (HSS) Levels During Liver Regeneration: Protective Function of HSS

Ethanol administration in rats induces liver damage and suppression of liver regeneration. To further understand the underlying mechanism, we investigated the effects of ethanol on hepatic stimulator substance (HSS) levels during liver regeneration caused by partial hepatectomy. The hepatotrophic action of HSS to ethanol-treated partially hepatectomized rats was also examined. Rats received repetitive ethanol or saline doses beginning 1 hr prior to 70% partial hepatectomy (PH), and the animals were killed at 16, 24, 32, 40, 48, and 60 hr after PH. Our results showed that ethanol inhibited hepatic regenerative capacity and prolonged liver regenerative process. HSS biological activity in ethanol-administered rats peaked at 48 hr after PH, in contrast to saline-treated ones where activity peaked at 24 hr. Additionally, exogenous HSS administration to ethanol-treated partially hepatectomized rats increased liver proliferating capacity and suppressed the elevation of serum ALT activity. These results showed that ethanol modifies the time course of HSS biological activity during the regenerating process. The observed suppression of HSS activity at 24 hr after PH was in relation with a reduction of DNA synthesis. Exogenous administration of HSS to ethanol-treated partially hepatectomized rats restored DNA synthesis and ameliorated serum AST levels, indicating that HSS could be used in the treatment of ethanol-induced hepatic failures.     

Effect of Hepatic Stimulator Substance (HSS) on Cadmium-Induced Acute Hepatotoxicity in the Rat Liver

The hepatoprotective effect of HSS against cadmium-induced liver injury was investigated. Rats were intoxicated with a dose of cadmium (3.5 mg/kg b.w.). The rats were treated with normal saline (group I) or HSS (100 mg protein/kg b.w.; group II) 2 hr later and killed at different time points. Hematoxylin-eosin (HE) sections were assessed for necrosis, apoptosis, peliosis, mitoses, and inflammatory infiltration. Serum enzyme activities were assayed. Apoptosis was quantified by the Tunel technique. Thymidine kinase activity and the rate of [3H]thymidine incorporation into DNA were also assayed. Necrosis, hepatocyte apoptosis, and peliosis were minimized in HSS-treated rats (group II). Nonparenchymal cell apoptosis and liver regeneration were not quantitively altered in the HSS-treated group, though the time profile was different. HSS protects hepatocytes against cadmium-induced necrosis, apoptosis, and peliosis. Apoptosis was the major type of cell death for nonparenchymal liver cells and strongly correlated with the extent of peliosis. Interactions between hepatocytes and nonparenchymal liver cells seem to be important for the genesis of hepatic trauma in acute cadmium hepatotoxicity.     

Hepatic stimulator substance administration ameliorates liver regeneration in an animal model of fulminant hepatic failure and encephalopathy

Abstract: Aims/Background: Hepatic stimulator substance (HSS) is a liver‐specific growth factor implicated in hepatocellular proliferation and hepatoprotection in models of acute liver injury. In the present study, we examined the effect of exogenous HSS administration on liver proliferating capacity and survival outcome in an experimental animal model of fulminant hepatic failure (FHF) and encephalopathy, induced by repeated injections of thioacetamide (TAA) in rats. Methods: Fulminant hepatic failure was induced in adult male Wistar rats by three consecutive intraperitoneal injections of TAA (400 mg/kg of body weight), at 24 h time intervals. The animals received intraperitoneally either a saline solution or HSS (50 mg protein/kg of body weight), 2 h after the second and third TAA injections. The animals were killed at 6, 12 and 18 h post the last injection of TAA. Results: Levels of liver enzymes and urea in serum, blood ammonia values, liver histology, stage of hepatic encephalopathy and survival were statistically significantly improved in TAA‐intoxicated and HSS‐treated rats compared to TAA‐intoxicated and saline‐treated ones. Furthermore, HSS ameliorated liver regenerative indices – DNA biosynthesis, thymidine kinase activity and hepatocyte mitotic activity – in a statistically significant manner. Conclusions: Our data suggest the beneficial effect of HSS administration in this animal model of FHF and encephalopathy, supporting evidence for a possible use of HSS as supportive therapy, by increasing hepatocellular proliferation, in management of FHF.     

Granulocyte-Colony Stimulating Factor Administration Ameliorates Liver Regeneration in Animal Model of Fulminant Hepatic Failure and Encephalopathy

It has previously been shown that recombinant granulocyte-colony stimulating factor (rG-CSF) accelerates and enhances hepatocyte proliferation in partially hepatectomized rats. In the present study, we examined the effect of rG-CSF administration on liver injury, regeneration, and survival outcome in an experimental rat model of fulminant hepatic failure (FHF) and encephalopathy induced by repeated injections of thioacetamide (TAA). FHF was induced in adult male Wistar rats by three consecutive intraperitoneal injections of TAA, at intervals of 24 hr. The animals were also injected with either saline or rG-CSF. Serum biochemical parameters and blood ammonia levels, liver histology, stage of hepatic encephalopathy, and survival were statistically significantly improved in TAA-intoxicated and rG-CSF-treated rats compared to TAA-intoxicated and saline-treated ones. Furthermore, rG-CSF not only ameliorated the histologically evident liver injury in a statistically significant manner but also enhanced the proliferative capacity of the hepatocytes. Our data confirm the beneficial effect of rG-CSF administration in this animal model of FHF and encephalopathy, supporting evidence for a possible use of rG-CSF as supportive therapy in the management of FHF.     

Stimulatory Effect of Epidermal Growth Factor on Liver Regeneration after Partial Hepatectomy in Rats

The effect of epidermal growth factor (EGF) on liver regeneration was investigated in rats subjected to partial hepatectomy. In a dose-response study EGF in doses of 6 and 24 nmol/kg X day increased liver regeneration after treatment for 48 h compared with controls, whereas a dose of 48 nmol/kg X day had no effect. In a subsequent study EGF, 6 nmol/kg X day, accelerated liver regeneration significantly after 36, 48, and 72 h of treatment. A possible influence of EGF on other hepatotrophic factors was investigated. No changes in the concentration of gastrin, insulin, or glucagon was found in portal venous blood. This study has shown that EGF in small doses can stimulate liver regeneration, whereas higher doses are ineffective. The study suggests that EGF should be regarded as a hepatotrophic factor.     

Hepatic Expression of Hepatocyte Growth Factor Gene mRNA in Acute Liver Failure

Hepatocyte growth factor plays a key role in liver regeneration but the role of liver in its synthesis in acute liver failure is unclear. We therefore measured hepatic expression of hepatocyte growth factor mRNA in this condition in comparison to H3 histone mRNA, a marker of cellular proliferation. Hepatocyte growth factor mRNA levels were quantified by specific RNase protection assay in nine patients with acute liver failure and found to be similar to those in six normal controls. Hepatocyte proliferation, as assessed by H3 histone mRNA expression, was not detected in normal liver but was present in six of nine patients with acute liver failure (P < 0.05) and was not correlated with expression of hepatocyte growth factor mRNA (r _s = –0.28). Liver is unlikely to be the source of the high serum hepatocyte growth factor levels observed in acute liver failure.     

Enhancement of Liver Regeneration by the Association of Hyptis pectinata with Laser Therapy

Since new molecules that normally would accelerate regeneration can also be potentialized by light, the use of new substances combined with laser therapy seems to be a natural type of experiment. Therefore, the purpose of this study was to assess the effects of Hyptis pectinata leaves on liver regeneration after partial hepatectomy (PH) associated with laser therapy. Twenty-four rats were divided into four groups—PH(control), PHL (laser therapy), PH200 (200 mg/kg of Hyptis pectinata), and PHL200 (200 mg/kg of the plant and laser)—which were submitted to 67% hepatectomy. Laser treatment consisted of focusing the light on the remaining liver after hepatectomy. The data analyzed were serum levels of aminotransferases, liver regeneration, and mitochondrial function. Group PH200 showed a statistically significant decrease in AST levels, and PHL200 disclosed an augmentation in ALT levels. The liver regeneration index was significantly increased in group PHL200. Concerning liver mitochondrial respiratory assay, groups PH200 and PHL200 showed lower state 3 levels than groups PH and PHL. Group PHL showed an increase in state 4 levels and a reduction in membrane potential and RCR. The present study shows that the association of the aqueous extract of Hyptis pectinata leaves at 200 mg/kg with intraoperative laser therapy can stimulate liver regeneration and cause a reduction in liver mitochondrial respiratory function without altering its phosphorylative activity.     

抗纤软肝颗粒对血小板源生长因子诱导的肝星状细胞增殖的影响

目的:研究抗纤软肝颗粒(KXR)对血小板源生长因子(PDGF)诱导的肝星状细胞(HSC)增殖的影响.方法:采用无血清培养,不同浓度的KXR温育HSC 24 h后,PDGF-BB(10 ng/ml)刺激24 h,再加入上述浓度的KXR 3 h后,又加入PDGF-BB(10 ng/ml)作用5 min,然后收集细胞.采用细胞计数法及流式细胞仪测定细胞增殖及细胞周期.结果:无血清培养显示该方对于PDGF诱导的细胞增殖具有抑制作用,并呈剂量依赖性(P<0.01).流式细胞术分析提示KXR能抑制PDGF诱导的HSC DNA合成期及合成后期和分裂期DNA百分含量,阻断细胞由静止期/DNA合成前期向DNA合成期转化,呈剂量依赖性(5 mg/ml组作用最显著,P<0.01).结论:KXR能抑制PDGF诱导的HSC增殖.     

Preventive Effect of FK 506 (Tacrolimus Hydrate) on Experimentally Induced Acute Liver Injury in Rats

The aim of the study was to investigate the effect of the immunosuppressant FK 506 (tacrolimus hydrate) on acute liver injury induced by Propionibacterium acnes and lipopolysaccharide (LPS). Acute liver injury was induced in male Wistar rats by injecting the animals with P. acnes (10 mg/rat), and administering LPS (10 g/rat) seven days later. One group was given FK 506 (1 mg/kg) 24 and 2 hr before administration of LPS, and the other group was given the same dose of saline. The 24-hr survival rate, serum alanine aminotransferase (ALT) concentration, and tumor necrosis factor (TNF) - mRNA and protein concentrations in the liver and spleen were then compared. Hepatic macrophages were also isolated from rats seven days after P. acnes injection, LPS, and FK 506 or saline were added to the culture supernatant, and TNF- production was studied. The 24-hr survival rate was 100% in the FK 506-treated group, in contrast with 16.6% in the saline group. Four hours after LPS injection, the serum ALT concentration was 755 ± 401 in the saline group versus 119 ± 42 units/ml (P < 0.01) in the FK 506-treated group. The serum TNF- concentration was lower in the FK 506-treated group (1419 ± 957 pg/ml) than in the saline group (9205 ± 2215) (P < 0.01). The mRNA and protein concentrations in the liver and spleen in the two groups did not differ significantly 1 hr after LPS injection but were significantly lower in the FK 506-treated group after 4 hr. FK 506 did not directly inhibit TNF- production by isolated cultured hepatic macrophages. FK 506 is unable to inhibit initial TNF- production by hepatic macrophages (or probably that by splenic macrophages either) stimulated by injection of LPS in P. acnes + LPS-induced acute liver injury. However, the immunosuppressant does limit hepatic damage by inhibiting subsequent aggravation of inflammation by the cytokine network.     

Post-hepatectomy haemorrhage: a definition and grading by the International Study Group of Liver Surgery (ISGLS)

BackgroundA standardized definition of post-hepatectomy haemorrhage (PHH) has not yet been established.MethodsAn international study group of hepatobiliary surgeons from high-volume centres was convened and a definition of PHH was developed together with a grading of severity considering the impact on patients' clinical management.ResultsThe definition of PHH varies strongly within the hepatic surgery literature. PHH is defined as a drop in haemoglobin level >3 g/dl post-operatively compared with the post-operative baseline level and/or any post-operative transfusion of packed red blood cells (PRBC) for a falling haemoglobin and/or the need for radiological intervention (such as embolization) and/or re-laparotomy to stop bleeding. Evidence of intra-abdominal bleeding should be obtained by imaging or blood loss via the abdominal drains if present. Transfusion of up to two units of PRBC is considered as being Grade A PHH. Grade B PHH requires transfusion of more than two units of PRBC, whereas the need for invasive re-intervention such as embolization and/ or re-laparotomy defines Grade C PHH.ConclusionThe proposed definition and grading of severity of PHH enables valid comparisons of results from different studies. It is easily applicable in clinical routine and should be applied in future trials to standardize reporting of complications.A proposed international definition and grading of severity of post hepatectomy haemorrhage which may enable better comparison of outcomes from future published studies     

Liver regeneration is not altered in patients with nonalcoholic steatohepatitis (NASH) when compared to chronic hepatitis C infection with similar grade of inflammation

Fatty infiltration is associated with an increased incidence of complications and mortality after liver resection and transplantation. The aim of this study was to document the regenerative response in patients with hepatic steatosis and mild inflammatory activity (NASH) and to identify potential levels of impaired regeneration. Ki-67 immunostaining was similar in patients with NASH (ages 44.6 ± 15 years, labeling index, 0.4 ± 0.3%) when compared to patients with chronic hepatitis C infection (ages 50.7 ± 17 years, labeling index; 0.4 ± 0.7%). The labeling index was not increased in patients with a higher level of inflammation, a higher level of fibrosis, and a higher level of fat in either study group. In conclusion, liver regeneration is not altered in patients with nonalcoholic steatohepatitis, suggesting that the delayed postoperative liver failure seen in these patients may be related to another mechanism.     

Detection of proliferating hepatocytes by immunohistochemical staining for proliferating cell nuclear antigen (PCNA) in patients with acute hepatic failure

ABSTRACT— This study evaluated whether liver regeneration could take place after massive or submassive necrosis of liver cells in 25 patients with several kinds of hepatic failure by immunohistochemical staining for proliferating cell nuclear antigen (PCNA). PCNA positivity was significantly higher (P<0.01) in the patients who survived than in the patients who died. Furthermore, PCNA-positive hepatocytes were recognized diffusely in the lobule of the liver in survivors. There was positive correlation between PCNA positivity and plasma concentration of AFP (α-fetoprotein), (r = 0.77, P<0.01). These results show that liver regeneration could take place after massive necrosis of liver cells in survivors from acute hepatic failure and that immunohistochemical staining for PCNA is useful for prognostic evaluation.     

扶正化瘀方对大鼠CCl4损伤肝Kupffer细胞功能的影响

目的：探讨扶正化瘀方对大鼠急性ＣＣｌ４损伤肝Ｋｕｐｆｆｅｒ细胞功能的影响。方法：体外分离培养大鼠急性ＣＣｌ４损伤肝Ｋｕｐｆｆｅｒ细胞,制备扶正化瘀方药物血清对其进行干预,观察药物对其ＰＤＧＦ和ＴＧＦβ活性以及对Ｋｕｐｆｆｅｒ细胞线粒体呼吸功能的影响。结果：药物组ＰＤＧＦ和ＴＧＦβ活性均显著降低（Ｐ〈０．０５）;药物组闰体呼吸功能明显降低（Ｐ〈０．０５）。结论：扶正化瘀方对大鼠急性ＣＣｌ４损伤肝Ｋｕｐｆｆｅｒ细胞活化有抑制作用,可能抑制肝星状细胞旁分泌激活,是该方抗肝纤维化的作用机理之一。     

Protective effect of some vitamins against the toxic action of ethanol on liver regeneration induced by partial hepatectomy in rats

AIM： To investigate the effects of vitamins （A, C and E） on liver injury induced by ethanol administration during liver regeneration in rats. METHODS： Male Wistar rats subjected to 70% partial hepatectomy were divided into five groups （groups 1-5）. During the experiment, animals of Group 1 drank only water. The other four groups （2-5） drank 30 mL of ethanol/L of water. Group 3 additionally received vitamin A, those of group 4 vitamin C and those of group 5 received vitamin E. Subsequently serum alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, albumin and bilirubin were measured colorimetrically. Lipid peroxidation （thiobarbituric-acid reactive substances, TBARS） both in plasma and liver was measured, as well as liver mass gain assessment and total DNA. RESULTS： Compared with sham group, serum AST and ALT increased significantly under ethanol treatment （43% and 93%, respectively, with P 〈 0.05）. Vitamin C and vitamin E treatment attenuated the ethanol-induced increases in ALT and AST activity. Ethanol treatment also decreased serum albumin concentration compared to sham group （3.1 ± 0.4 g/dL vs 4.5 ± 0.2 g/dL; P 〈 0.05）. During liver regeneration vitamins C and E significantly ameliorated liver injury for ethanol administration in hepatic lipid peroxidation （4.92 nmol/mg and 4.25 nmol/mg vs 14.78 nmol/mg, respectively, with P 〈 0.05）. In association with hepatic injury, ethanol administration caused a significant increase in both hepatic and plasma lipid peroxidation. Vitamins （C and E） treatment attenuated hepatic and plasma lipid peroxidation. CONCLUSION： Vitamins C and E protect against liver injury and dysfunction, attenuate lipid peroxidation, and thus appear to be significantly more effective than vitamin A against ethanol-mediated toxic effects during liver regeneration.     

热毒清对内毒素所致DIC大鼠肝损伤的防护作用

目的:观察热毒清抗内毒素DIC大鼠肝损伤的作用。方法:用内毒素诱发大鼠DIC模型观察热毒清对肝组织病理学和丙氨酸氨基转移酶(ALT)的影响。结果:和模型组相比,热毒清组的ALT水平明显降低,差别有显著意义(P<0.01)。病理学改变也明显减轻。结论:热毒清对内毒素DIC大鼠肝损伤有一定的改善作用。     

A Case of Massive Hepatic Necrosis (Scarred Liver) with Elevated α-Fetoprotein and Seroconversion from HBe Ag to HBe Ab

A 39-year-old male with general malaise visited our clinic with high levels of aminotransferases (AST 776 IU/1, ALT 1,069 IU/I) and a prolonged prothrombin time. Serum-fetoprotein (AFP) level was markedly elevated (2,214 ng/ml) and human hepatocyte growth factor (hHGF)was also high (0.81 ng/ml). Hepatocellular carcinoma was found to be negative from imaging techniques. AFP bands separated by lectin affinity electrophoresis had a benign pattern. Laparoscopic diagnosis was scarred liver and histologically massive hepatic necrosis with a strongly positive AFP stain. Levels of hHGF and AFP fell into a normal range after treatments with glucagon-insulin and prostaglandin Ei. At second laparoscopy, regenerated nodules were confirmed by improved staining with Indocyanine Green. Hepatocyte regeneration was also histologically proven with a weakly positive AFP stain. The proliferating cell nuclear antigen (PCNA) was stained, and hepatocytes with PCNA positive nuclei were not seen at the time of the first laparoscopy, whereas hepatocytes with PCNA positive nuclei were found to be many at the time of the second laparoscopy. Seroconversion from HBe Ag to HBe Ab was observed 4 months after the maximum rise of AFP (6,341 ng/ml), when levels of ALT and DNA polymerase were within normal ranges. This is a patient with a marked rise in his serum level of AFP in association with seroconversion from HBe Ag to HBe Ab, and the elevation of AFP in the present case was thought to reflect severe liver injury with concomitant regenerative activities of injured hepatocytes rather than simple hepatocyte regeneration after hepatic necrosis was resolved.     

The Effect of Taurine on Hepatic Steatosis Induced by Thioacetamide in Zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>)

Background  

Nonalcoholic fatty liver disease is one of the most prevalent forms of chronic liver disease in the Western world. Taurine is a conditionally essential amino acid in humans that may be a promising therapy for treating this disease.     

阿魏酸钠对局灶性脑缺血致多器官功能障碍综合征大鼠模型脑和肺及肾Bax和Bcl-2表达的影响

目的研究阿魏酸钠对大鼠急性局灶性脑缺血再灌注后脑、肺和肾Bax、Bcl-2表达的影响，探讨阿魏酸钠对急性局灶性脑缺血致多器官功能障碍综合征（MODS）的治疗作用。方法参照Longa等的线栓法采用阻断大鼠大脑中动脉（MCAO）后再灌注法建立大鼠局灶性脑缺血致MODS模型，按随机原则将66只Wistar大鼠分为正常组（n=6）、假手术组（n=6）及手术组（缺血再灌注后2h、6h、12h、24h、48h、72h、5d，7个亚组），阿魏酸钠治疗组（n=6），0．9％氯化钠溶液组（n=6），应用免疫组织化学SP染色技术检测脑、肺、肾Bax、Bcl-2的蛋白表达并计数，计算出平均阳性率（LI）。结果（1）大鼠脑缺血后各时相点的脑、肺和肾组织均有不同程度的病理损害；（2）全身炎症反应综合征（SIRS）的发生率为100．0％，MODS的发生率为57、1％；（3）再灌注后72h 3组大鼠Bax、Bcl-2在脑、肺、肾的阳性细胞数间差异均有显著性意义（P〈0.05），且两两比较阿魏酸钠组与正常组、0．9％氯化钠溶液组间差异均有非常显著性意义（P〈0．01）。结论SIRS和Bax、Bcl-2表达失衡是脑缺血致MODS的可能机制，阿魏酸钠通过重建Bax、Bcl-2表达平衡起到治疗作用。     

Metabolic-associated fatty liver disease: From simple steatosis toward liver cirrhosis and potential complications. Proceedings of the Third Translational Hepatology Meeting,organized by the Spanish Association for the Study of the Liver (AEEH)

This is a meeting report of the 3rd Translational Hepatology Meeting held in Alicante, Spain, in October 2021. The meeting, which was organized by the Spanish Association for the Study of the Liver (AEEH), provided an update on the recent advances in the field of basic and translational hepatology, with a particular focus on the molecular and cellular mechanisms and therapeutic targets involved in metabolic-associated fatty liver disease (MAFLD), metabolic-associated steatohepatitis (MASH), cirrhosis and end-stage hepatocellular carcinoma (HCC).