Hepatic platelet accumulation in Fas-mediated hepatitis in mice

Platelets are reported to be causally involved in experimental hepatitis. Jo2, an agonistic anti-Fas antibody, induces hepatitis in mice. We examined the in vivo behaviors of platelets in mice injected with this antibody (analyzed by measuring 5-hydroxytryptamine, a constituent of platelets). We found that Jo2 induces platelet accumulation predominantly in the liver, and that this hepatic platelet accumulation (HPA) precedes the increases in hepatitis markers (alanine- and asparagine-aminotransferases [ALT and AST]). By electron microscopy, we detected entry of platelets into hepatocytes, and also evidence of apoptosis among hepatocytes. A caspases-3/6/7/8/10 inhibitor prevented the Jo2-induced HPA and hepatitis. In platelet-depleted mice, contrary to our expectations, the Jo2-induced hepatitis was not reduced, and actually the increase in AST was significantly augmented, although the survival time of mice given a lethal dose of Jo2 was significantly increased (nearly doubled). Interestingly, prior induction of HPA by a low dose of lipopolysaccharide markedly reduced Jo2-induced hepatitis. Jo2 also induced HPA and hepatitis in mice deficient in both IL-1 and TNFα, although Jo2 increased the blood level of TNFα in wild-type mice. These results suggest that in Jo2-induced hepatitis: (i) platelets accumulate predominantly in the liver as a result of hepatic lesions, and that this precedes the release of transaminases from hepatocytes, and (ii) IL-1 and TNFα are not essential for Jo2-hepatitis. We hypothesize that platelet accumulation in the liver may, contrary to our expectations, be protective when the hepatitis is local or not severe, but harmful when hepatitis is severe.     

Protective effect of Acanthopanax gracilistylus-extracted Acankoreanogenin A on mice with fulminant hepatitis

The release of pro-inflammatory cytokines in both acute (IL-1β and TNF-α) and chronic [high mobility group box 1 protein (HMGB1)] phases, is thought to play important roles in the development of fulminant hepatitis (FH). Triterpenoid Acankoreanogenin A (AA) which is extracted from the leaves of the Acanthopanax gracilistylus W.W. Smith (AGS) has shown its inhibiting effect on TNF-α, IL-1β and HMGB1 release in vitro in our preliminary experiments. In present study, we investigated the effect of AA on mice with fulminant hepatitis in vivo. Fulminant hepatitis mice model was established by intraperitoneally injecting galactosamine (GalN) and lipopolysaccharide (LPS). The levels of serum of TNF-α, IL-1β, ALT, AST and HMGB1 from AA-treated mice were measured at different time points. Our results demonstrated that pre-treatment of mice with AA markedly reduced the serum levels of TNF-α, IL-1β, HMGB1, ALT and AST with the improvement in histological features. And the survival rate from AA-treated fulminant hepatitis mice was increased. Furthermore, delayed administration of AA after peak occurrence of the early pro-inflammatory cytokines still endowed significant protection against GalN/LPS-induced lethality. The post-treatment of AA could significantly attenuate the release of HMGB1, but not the TNF-α and IL-1β. These results indicate that AA inhibits the systemic release of pro-inflammatory cytokine HMGB1, and dose-dependently rescue the mice from lethal GalN/LPS-induced fulminant hepatitis, which suggests this component as a candidate therapy for fulminant hepatitis.     

The antibiotic minocycline prevents methamphetamine-induced rewarding effects in mice

Repeated use of methamphetamine (METH) causes dependence in humans, and to date, there are no effective medication treatments for METH addiction. We previously reported that the antibiotic minocycline attenuated behavioral abnormalities (hyperactivity and behavioral sensitization) and dopaminergic neurotoxicity in mice and monkeys, after the administration of METH. In this study, we examined the effect of minocycline on METH-induced rewarding effects in mice using the conditioned place preference (CPP) paradigm. Minocycline (40 mg/kg, IP) significantly attenuated METH (1.0 mg/kg, SC)-induced place preference in mice. In vivo microdialysis experiments using free-moving mice, showed that minocycline (40 mg/kg, IP) significantly attenuated the increased extracellular dopamine (DA) levels within the nucleus accumbens, typically seen after the administration of METH (1.0 mg/kg, SC). These findings suggest that minocycline may block METH-induced rewarding effects by down regulating extracellular DA levels in the nucleus accumbens of mice. This would make minocycline a potential therapeutic drug for the treatment of METH induced disorders.     

Improvement by minocycline of methamphetamine-induced impairment of recognition memory in mice

Introduction Cognitive deficits are a core feature of patients with schizophrenia and methamphetamine (METH) psychosis. We have recently found that repeated METH treatment (1 mg/kg, s.c.) in mice, which induces behavioral sensitization, impairs long-term recognition memory in a novel object recognition test (NORT) and that the impairment is ameliorated by clozapine, but not haloperidol. Recent studies indicate that minocycline, a second-generation tetracycline, has potent neuroprotective effects in various animal models of neurological diseases. Objectives In the present study, we investigated the effect of minocycline on learning and memory in the NORT and behavioral sensitization in mice that had been administered METH for 7 days. Results When minocycline (20–40 mg/kg) was administered intraperitoneally once a day for seven consecutive days to mice that had previously been treated with METH for 7 days, it ameliorated the METH-induced impairment of recognition memory in a dose-dependent manner, although the same treatment with minocycline had no effect on behavioral sensitization to METH. The administration of minocycline, together with METH, inhibited the development of METH-induced behavioral sensitization. The improvement in memory caused by minocycline was associated with an amelioration of the novelty-induced activation of extracellular signal-regulated kinase 1/2 in the prefrontal cortex of METH-treated mice. Conclusions These results suggest that minocycline is useful for the treatment of cognitive deficits in patients with METH psychosis or schizophrenia.     

促肝细胞生长素治疗重肝倾向型肝炎的临床研究

目的：研究促肝细胞生长素治疗重肝炎的临床疗效。方法：选择70例重肝倾向肝炎病人,随机分为治疗组及对照组,每组各35例。对照组应用保肝退黄支持治疗,治疗组在对照组基础上每日加用促肝细胞生长素。结果;治疗组有效率77.14%,对照组有效率54.2%,两组比较有显著性差异（P＜0.05）。重症肝炎的发生率;对照组28.5%,治疗组为14.29%,两者比较有高显著性差异（P＜0.01）。结论：促进细胞生长素对重肝倾向型肝炎的治疗有效,并可降低重症肝炎的发生率。     

Effects of serum lipid content on the binding of minocycline.

Minocycline was added to normal and hyperlipemic serum samples in concentrations of 1 approximately 10 mcg/ml. These specimens had similar protein contents. Chemically extractable minocycline was quantitated fluorometrically. Hyperlipemic serum (cholesterol 480 mg/100 ml; triglycerides 321 mg/100 ml) yielded an average of 50% less minocycline than did normal serum (cholesterol 170 mg/100 ml; triglycerides 114 mg/100 ml). When ultrafiltrates of serum containing 6, 12 and 20 mcg/ml minocycline were assayed microbiologically, it was evident that variations in serum triglyceride and cholesterol levels did not alter the ratio of bound to free drug. Minocycline appears to be reversibly associated with, and/or soluble in, triglyceride-cholesterol components of serum.     

Antidiabetic drug metformin alleviates endotoxin-induced fulminant liver injury in mice

Metformin is a first-line antidiabetic drug in type 2 diabetes for its hypoglycemic activity, but recently researches also revealed the anti-inflammatory properties of metformin. In the present study, the pharmacological efficiency of metformin in lipopolysaccharide (LPS)-induced hepatic injury in D-galactosamine (D-Gal)-sensitized mice was investigated. We found that pretreatment with metformin significantly decreased serum ALT and AST levels in LPS/D-Gal-exposed mice. These were accomplished with improved histological alterations in liver sections, decreased myeloperoxidase (MPO) activity, reduced malondialdehyde (MDA) content in liver homogenates and increased survival rate of experimental animals. Metformin also markedly reduced hepatic TNF-α mRNA content and blood TNF-α level. Additional experiment showed that metformin significantly attenuated LPS/D-Gal-induced hepatic apoptosis as evidenced by decreased caspase activities in liver tissues and reduced number of TUNEL-positive cells in liver sections. Furthermore, therapeutic administration of metformin after LPS/D-Gal challenge also improved the survival rate of experimental animal. These results indicated that the hypoglycemic reagent metformin could also provide therapeutic benefits in endotoxin-induced hepatic injury, suggesting its pharmacological potential in inflammation-base disorders.     

米诺环素调控小胶质细胞激活对PC12细胞生长的影响

目的探讨米诺环素抑制小胶质细胞激活对PC12生长和凋亡的影响。方法 BV2细胞分为对照组、LPS组、LPS加米诺环素组;以LPS刺激激活小胶质细胞系BV2细胞,用米诺环素干预BV2细胞的激活;将各组BV2细胞与PC12细胞进行共培养24h;酶联免疫吸附法检测共培养体系细胞上清液TNF-α、IL-1β的含量,MTT法检测PC12细胞生存率。结果 BV2细胞与PC12细胞共培养24h后,LPS组上清液炎症因子TNF-α、IL-1β表达明显升高,PC12细胞存活率下降,米诺环素能抑制以上趋势。结论 MG激活对PC12细胞存在明显损伤效应,米诺环素可保护MG介导的PC12细胞损伤,可能与通过下调炎症表达有关。     

The role of glucocorticoids in the treatment of fulminant hepatitis induced by dacarbazine

Dacarbazine (DTIC) is commonly used for the treatment of malignant melanoma and Hodgkin's disease. A very rare complication of this cytotoxic agent is acute vascular hepatic damage, e.g. veno-occlusive disease or Budd-Chiari syndrome. The pathophysiological mechanism involved seems to be an immune reaction. This complication is frequently fatal. We report a patient who developed severe hepatic failure following DTIC treatment who responded favorably to treatment with glucocorticosteroid.     

胸腺肽α1治疗慢性重型肝炎并自发性腹膜炎疗效观察

目的探讨胸腺肽α1治疗慢性重型肝炎并自发性腹膜炎的作用机制及疗效。方法60例慢性重型肝炎并发腹膜炎患者,按入院单双号随机分成治疗组和对照组各30例,对照组行常规综合治疗;治疗组在对照组的基础上加用胸腺肽α1皮下注射,1．6mg／次,2次／d,连用3d后改为1次／d,疗程2周,观察两组治疗效果及T淋巴细胞亚群变化情况。结果治疗组临床症状缓解时间、肝功能恢复时间均明显短于对照组（均P〈0．05）;治疗组显效10例,有效17例,无效3例,总有效率90．0％,与对照组的5例、14例、11例、63．3％比较,差异有统计学意义（P〈0．05）;治疗组CD3^＋、CD4^＋、NK细胞和CD4^＋／CD8^＋治疗后明显高于治疗前（均P〈0．05）,对照组治疗前后各项免疫指标无明显变化。结论胸腺肽α1可显著改善慢性重型肝炎并发自发性腹膜炎患者的免疫功能,从而提高治疗效果。     

重型肝炎并发多器官功能衰竭分析

目的探讨重型肝炎与器官功能衰竭（MOF）的关系。方法通过对296例重型肝炎并发多器官功能衰竭临床资料分析。结果重型肝炎并发多器官衰竭病死率为68．2％，器官受累数目越多，预后越差，衰竭器官4个以上病死率均为100％。其主要病因是各种感染及内毒素。结论加强原发病综合治疗措施，控制感染，可改善预后。     

米诺环素的药效学研究

国产米诺环素（ｍｉｎｏｃｙｃｌｉｎｅ，ＭＩＮＯ）的体内外抗菌活性研究结果表明：ＭＩＮＯ是一个对革兰氏阳性菌和革兰氏阴性菌均有较强抗菌活性的广谱抗生素，对金葡球菌、表葡球菌的抗菌活性优于四环素（ＴＣ）、青霉素（ＰＣＧ）、麦迪霉素（ＭＤＭ）、庆大霉素（ＧＭ）和依诺沙星（ＥＮＸ），并对耐四环素金葡球菌和表葡球菌有很强的抗菌活性。ＭＩＮＯ对化脓性链球菌的抗菌活性优于ＴＣ、ＭＤＭ、ＧＭ、ＥＮＸ，弱于ＰＣＧ；对大肠杆菌、肺炎克雷伯氏菌的抗菌活性优于ＴＣ、ＧＭ，弱于ＥＮＸ；对变形杆菌的抗菌活性优于ＴＣ，弱于ＧＭ、ＥＮＸ。口服ＭＩＮＯ对金葡球菌感染小鼠的ＥＤ５０为０．９９ｍｇ／ｋｇ，优于强力霉素（ＤＯＸＹ）和ＴＣ；对大肠杆菌感染小鼠的ＥＤ５０为１０．９６ｍｇ／ｋｇ，优于ＴＣ，与ＤＯＸＹ的ＥＤ５０无显著性差异；对耐ＴＣ金葡球菌感染小鼠的ＥＤ５０为０．６５ｍｇ／ｋｇ，明显优于ＤＯＸＹ和ＴＣ。杀菌试验表明：ＭＩＮＯ在２～４ＭＩＣ时，对金葡球菌和大肠杆菌具有杀菌作用。     

Protective effects of agmatine against d-galactosamine and lipopolysaccharide-induced fulminant hepatic failure in mice

Fulminant hepatic failure (FHF) is a life-threatening syndrome characterized by massive hepatic necrosis and high mortality. There is no effective therapy for the disease other than liver transplantation. This study aimed to investigate the effect of agmatine, inducible nitric oxide synthase (iNOS) inhibitor, on d-galactosamine and lipopolysaccharide (GalN/LPS)-induced FHF in mice and explore its possible mechanism(s). Male Swiss albino mice were injected with a single dose agmatine (14 mg/kg, IP) 8 h prior to challenge with a single intraperitoneal injection of both GalN (800 mg/kg) and LPS (50 μg/kg). Agmatine significantly attenuated all GalN/LPS-induced biochemical and pathological changes in liver. It prevented the increase of serum transaminases and alkaline phosphatase (ALP). In addition, agmatine markedly attenuated GalN/LPS-induced necrosis and inflammation. Agmatine significantly reduced oxidative stress and enhanced antioxidant enzymes. Importantly, agmatine decreased total nitric oxide (NO) and pro-inflammatory cytokine, tumor necrosis factor-alpha (TNF-α). These findings reveal that agmatine has hepatoprotective effects against GalN/LPS-induced FHF in mice that may be related to its ability to suppress oxidative stress, NO synthesis and TNF-α production. Therefore, agmatine may serve as a novel therapeutic strategy for hepatic inflammatory diseases.     

地塞米松治疗有重症倾向的慢性乙型肝炎的临床观察

目的观察地塞米松对有重症倾向的慢性乙型肝炎患者的疗效及预后。方法将120患者分为3组,A组保肝抗炎、保护胃黏膜、预防感染、对症支持等综合治疗;B组在A组基础上加用地塞米松10mg冲击治疗3～5d后停用;C组在A组治疗基础上地塞米松3mg,间隔48～72h再使用,疗程6周。结果治疗6周,B组、C组总胆红素水平、消化道症状改善、重症肝炎发生率与A组比较有统计学意义(P<0.05)。C组并发症与B组对比差异显著(P<0.05)。结论有重症倾向的慢性乙型肝炎患者采用地塞米松冲击治疗或小剂量间断使用可以早期缓解消化道症状、降低重症肝炎发生率,但小剂量间断使用疗法更易出现并发症,地塞米松短期冲击治疗更安全。     

桔梗提取物对D-氨基半乳糖/内毒素诱导暴发性肝衰竭的影响

目的探讨桔梗提取物对D-氨基半乳糖/内毒素诱导的小鼠暴发性肝衰竭的预防作用。方法采用D-氨基半乳糖/内毒素诱导的小鼠暴发性肝衰竭模型,桔梗提取物小剂量组给予桔梗提取物150 mg/kg,桔梗提取物大剂量组给予桔梗提取物300 mg/kg,阳性对照组给予谷胱甘肽300 mg/kg,给药2次。末次给药1 h后除正常组外各组均腹腔注射D-氨基半乳糖(700 mg/kg)和内毒素(10μg/kg),8 h后,颈动脉取血,测定血清中丙氨酸氨基转肽酶(ALT)和天门冬氨酸氨基转移酶(AST)含量,取肝脏,液氮保存。结果模型组小鼠肝损伤严重,ALT、AST较高,给药组AST及ALT降低。结论桔梗提取物能够减轻D-氨基半乳糖/内毒素诱导的小鼠暴发性肝衰竭,对肝脏起到保护作用。     

Metabolism of minocycline in humans

Metabolites of tetracycline antibiotics have not previously been isolated from human excreta. It is now demonstrated that minocycline is the first tetracycline to be metabolized in man. A two-step liquid chromatographic procedure was used to isolate three metabolites from human urine. The principal metabolite was tentatively identified as 9-hydroxyminocycline by mass spectral and spectrophotometric analysis and comparison with a synthetic compound. The latter was prepared by incubation of minocycline in a modified Udenfriend system, simulating an enzymatic oxygenase. Two other major metabolites are probably mono-N-demethylated derivatives. Substantial amounts of 4-epiminocycline, which probably resulted from minocycline epimerization rather than biotransformation, were also present.