Role of glucose-regulated protein 78 in pulmonary microvascular remodeling during development of rat hepatopulmonary syndrome

AIM：To explore the mechanisms of pulmonary microvascular remodeling induced by glucose-regulated protein 78 (GRP78) in the development of hepatopulmonary syndrome (HPS) in rats. METHODS：The Wistar rats were randomly divided into HPS groups at the 4th, 6th and 8th weeks, and normal control groups at the corresponding time points. The rat model of HPS produced in the process of liver cirrhosis was induced by employing multiple pathogenic factors to the animals. The rats in normal control group were designed by feeding with standard diet and mineral water. The expression of GRP78, factor Ⅷ- related antigen (FⅧ-RAg), C/EBP homologous protein (CHOP, also called growth arrest and DNA damage-inducible protein 153, GADD153), caspase-12, Bcl-2 and nuclear factor (NF)-κB in the lung tissues were measured by immunohistochemistry. The expression of VEGF at mRNA and protein levels in the lungs was measured by the methods of RT-PCR and Western blotting, respectively. RESULTS：The expression of GRP78, FⅧ-RAg,VEGF and Bcl-2 proteins was gradually increased with the HPS development. The protein expression of NF-κB was also gradually increased, especially in nucleus. GRP78 protein in the lung was positively correlated with the expression of FⅧ-RAg and VEGF, but negatively correlated with the expression of CHOP/GADD153 and caspase-12. The protein levels of GRP78 and FⅧ-RAg, and VEGF at both mRNA and protein levels were higher, and the protein levels of CHOP/GADD153 and caspase-12 were lower in the rats with HPS at every time point than those in normal control rats (P<0.05). CONCLUSION：Overexpression of GRP78 in the lung may be the critical pathogenesis of HPS by promoting cell survival and proliferation, and inhibiting cell apoptosis, thus leading to pulmonary microvascular remodeling.     

Proliferation-inhibitory and apoptosis-inducing effects of cinnamic acid combined with cisplatin on human hepatocellular carcinoma cell line MHCC97

AIM：To investigate the effects of cinnamic acid (CA) combined with cisplatin on the proliferation and apoptosis of human hepatocellular carcinoma cell line MHCC97. METHODS：Human hepatocellular carcinoma cell line MHCC97 was culured and divided into CA group, cisplatin group, CA+cisplatin group and control group. MTT assay, inverted microscopy, annexin V-FITC/PI staining and flow cytometry were applied to identify the viability, morphology and apoptosis of the cells. The apoptosis-related signaling protein caspase-3 was detected by Western blotting. RESULTS：CA and cisplatin either alone or in combination significantly inhibited the proliferation and induced obvious apoptosis of MHCC97 cells, while CA alone or combined with cisplatin had no significant inhibitory effect on normal human liver L-02 cells. The rates of mid-and late apoptosis or necrosis were higher in cisplatin group than that in CA group or combination group, but the early apoptotic rate was just the opposite. Pro-apoptotic activity in combination group was much stronger than that in CA group or cisplatin group at lower concentration, and combination group promoted apoptosis and decreased the cytotoxic side effects of cisplatin. CA and cisplatin either alone or in combination also up-regulated the cleaved caspase-3 expression in a time-dependent manner, and the effects in CA group and combination group were higher than that in cisplatin group. CONCLUSION：CA and cisplatin either alone or in combination inhibit the growth of MHCC97 cells by inducing apoptosis, and the activation of caspase-3 may play important roles in these processes.     

Effects of Nrf2 overexpression on proliferation,activation and collagen type I synthesis in cultured hepatic stellate cells stimulated by ethanol

AIM：To investigate the effects of nuclear factor E2-related factor 2 (Nrf2) overexpression on the proliferation, cell cycle distribution, collagen type I (Col I) synthesis and alpha-smooth muscle actin (α-SMA) expression in cultured hepatic stellate cell line HSC-T6 stimulated by ethanol. METHODS：Cultured HSC-T6 cells were transfected with pEGFP-Nrf2 or pEGFP-N1 (empty vector) plasmid by liposome transient transfection. The cells were divided into control group, ethanol group, ethanol+pEGFP-Nrf2 group and ethanol+pEGFP-N1 group. The mRNA expression of Nrf2, α-SMA and Col I was determined by RT-PCR, and their protein expression was detected by Western blotting. Cell proliferation was assessed by MTT assay, and cell cycle was detected by flow cytometry. RESULTS：The pEGFP-Nrf2 plasmid was successfully transfected into HSC-T6 cells, and the mRNA and protein expression of Nrf2 was higher than other three groups 48 h after transfection (P<0.05). Compared with control group, the cell proliferation and the mRNA and protein expression of α-SMA and Col I in ethanol group and ethanol+pEGFP-N1 group were significantly increased (P<0.05), and the numbers of HSC-T6 cells were decreased in G1 phase and increased in S phase (P<0.05), without significant differences between the two groups (P>0.05). Meanwhile, the cells in ethanol+pEGFP-Nrf2 group showed significantly decreased proliferation level, down-regulated mRNA and protein expression of α-SMA and Col I, higher numbers in G1 phase and lower numbers in S phase compared with ethanol group and ethanol+pEGFP-N1 group (P<0.05). CONCLUSION：Nrf2 overexpression could significantly down-regulate the expression of α-SMA and Col I and cause G1/S phase arrest in HSC-T6 cells cultured with ethanol, thus inhibiting the proliferation and activation of the cells.     

Role of hydrogen sulfide in alleviation of liver injury by mesenteric lymph drainage in hemorrhagic shock rats

AIM：To investigate the role of hydrogen sulfide (H2S) in alleviation of liver injury by mesenteric lymph drainage in hemorrhagic shock rats. METHODS：A hemorrhagic shock model was established in male Wistar rats. DL-propargylglycine (PPG), an inhibitor of cystathionine γ-lyase (CSE) which is a synthase of H2S, or sodium hydrosulfide (NaHS), a donor of H2S, was administered to the hemorrhagic shock rats with mesenteric lymph drainage. The rats were randomly divided into sham, shock, shock+drainage, shock+drainage+PPG (45 mg/kg, ip, 0.5 h before hemorrhage) and shock+drainage+NaHS (28 μmol/kg, ip, 0.5 h before hemorrhage) groups. Fluid resuscitation was performed 1 h after hypotension, and then mesenteric lymph was drained in the rats of shock+drainage, shock+drainage+PPG and shock+drainage+NaHS groups for 3 h. The hepatic histomorphology was observed. The biochemical indexes of hepatic function in plasma, and H2S, CSE, Toll-like receptor 4 (TLR4), interleukin (IL)-10, IL-12 and tumor necrosis factor α (TNF-α) in hepatic homogenate were also examined. RESULTS：The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bile acid (TBA) in plasma, and H2S, CSE, TLR4, IL-10, IL-12 and TNF-α in hepatic homogenate in shock group were significantly higher than those in sham group. Mesenteric lymph drainage obviously reduced these indexes in shock rats, except for TLR4. PPG further decreased these indexes except for CSE, while NaHS increased these indexes except for TBA and CSE. Morphological observation showed that liver injury appeared in the rats from shock and shock+drainage+NaHS groups, and there was nearly normal hepatic structure in the rats from sham, shock+drainage and shock+drainage+PPG groups. CONCLUSION：The mechanism of mesenteric lymph drainage alleviating liver injury in hemorrhagic shock rats is related to reducing the production of H2S and alleviating the H2S-mediated inflammation.     

Effect of exposure to a country liquor (Toddy) during gestation on lipid metabolism in rats

The objective of this study was to determine theeffects of country liquor Toddy and its equivalentquantity of ethanol on lipid metabolism duringgestation in rats. Female rats weighing an average of125 g were exposed to Toddy (24.5 ml/body weight/day)and ethanol (0.52 ml/kg body weight/day) for 15 daysbefore conception and throughout gestation. On the19th day of gestation, altered liver function andhyperlipidemia was seen in both the treated groups.Altered liver function was evidenced by the increasedactivity of alcohol dehydrogenase, aldehydedehydrogenase, glutamic oxaloacetic transaminase oraspartate amino transferase (GOT), glutamic pyruvictransaminase or alanine amino transferase (GPT) andgamma glutamyl transpeptidase (GGT). Hyperlipidemiawas caused by increased biosynthesis and decreaseddegradation of lipids. The incorporation of ¹⁴Cacetate in lipids and activities of HMG CoA reductaseand lipogenic enzymes were elevated and activity ofLPL and bile acids contents were decreased. Toddytreated rats were more severely affected than thosereceiving an equivalent quantity of ethanol. Toddyseemed to potentiate the toxicity induced by alcoholindicating the role of the nonethanolic portion.Hepatic functions were also affected.     

饲料中添加发酵中药对文昌鸡肝毒性研究

中药渣经微生物发酵可减少抗营养因子,增强功效,试验旨在研究饲料添加发酵中药渣对文昌鸡肝脏毒性影响。试验选用体重、日龄相近文昌鸡(43日龄,母)192只,随机分为4组,每组8个重复,每个重复6只。发酵中药渣添加水平为0%、0.5%、1.0%、1.5%,试验为期4周。结果显示,与组1相比,2、3、4组血清中谷草转氨酶(AST)、总胆红素(TBIL)、葡萄糖(GLU)含量均显著降低(P<0.05)。与组1相比,2、3组肝脏组织AST含量显著升高(P<0.05);3组谷丙转氨酶(ALT)含量显著升高(P<0.05),4组ALT含量显著降低(P<0.05);2、3、4组碱性磷酸酶(AKP)含量显著升高(P<0.05);3、4组肝脏组织中GLU和总蛋白(TP)显著升高(P<0.05)。表明,饲料中添加发酵中药渣可减少文昌鸡肝细胞损伤,促进肝脏的合成代谢和贮备功能,其中以1.0%添加量最适。     

世界唯一棕白色大熊猫“丹丹”部分组织的光镜观察

对世界唯一一例人工圈养的棕白色大熊猫“丹丹”的肝胀、脾胀、子宫、癌变皮和口皮部分组织进行光镜观察。结果表明:肝窦和脾窦不同程度扩张,其中有大量红细胞和枯否氏细胞,可见大量黄褐色的含铁血黄素颗粒。部分肝细胞变性、坏死;大量肝细胞肿胀,胞浆内可见许多大小不一的脂肪空泡;肝细胞胞核浓缩、碎裂、溶解甚至消失。白髓淋巴小结里淋巴细胞增多。子宫的固有膜由富有血管的胚性结缔组织构成。癌变皮的棘细胞出现不典型增生,未见恶性细胞团向深层组织生长浸润。口皮的棘细胞未出现增生。     

内毒素诱导的山羊红细胞膜和肝线粒体膜Ca2+-ATP酶活性的改变及654-2的保护作用

 通过静脉注射的方法研究了大肠杆菌内毒素(ET)对山羊红细胞膜和肝粒体膜损伤的膜Ca²⁺-ATP酶活性的变化及654-2(山莨菪碱)的保护效应。结果表明,ET处理组(ⅱ组)在1h,3h红细胞膜Ca²⁺-ATP酶活性高于对照组(iv组)(p<0.05),5h有下降趋势且持续到9h.12h开始回升并低于iv组(p<0.01,p<0.05).654-2处理组(ⅲ组),Ca²⁺-ATP酶活性除在9h、12h与iv组,在3h与ⅱ组差异不显着(p>0.05)外,均高于iv组和ⅱ组(p<0.01,p<0.05).在肝线粒体中,ⅱ组Ca²⁺-ATP酶活性在5h、12h均低于iv组和ⅱ组(p<0.05,p<0.01),ⅲ组Ca²⁺-ATP酶活性均高于iv组,但12h与iv组差异不显着(p>0.05).结果提示,ET具有诱导膜Ca²⁺-ATP酶活性先激活后顿抑的效应,而654-2有明显保护膜Ca²⁺-ATP酶活性的作用。     

鲈鱼对7种饲料原料的表观消化率及其对肝脏、肠道组织结构的影响

试验以Cr2O3为指示物,以70%基础饲料和30%的待测饲料原料组成试验饲料,在室内流水养殖系统中,采用虹吸法收集粪便,测定了初始体重为30±2.3 g的鲈鱼Lateolabrax japonicus对白鱼粉、血粉、虾糠、羽毛粉、双低菜粕、高筋粉和米糠中干物质、粗蛋白和能量的表观消化率.试验结果表明,鲈鱼对不同饲料原料的干物质表观消化率为98.71%～41.84%.其中白鱼粉为98.71%,显著高于其他各组(P<0.05).鲈鱼对白鱼粉和米糠的蛋白表观消化率很高,均在98%以上,双低菜粕也较高,为86.86%,血粉、羽毛粉和高筋粉在62.94%～71.08 9,6之间,虾糠仅为45.01%,显著低于其他各组(P<0.05).鲈鱼对不同饲料原料的能量表现消化率为95.24%～65.54%,对白鱼粉和高筋粉在90%以上,对血粉、羽毛粉、双低菜粕和米糠的能量表观消化率在74%以上.进行70 d的饲养实验后,取鲈鱼肝脏和肠道组织制作切片.观察到血粉、羽毛粉、双低菜粕、高筋粉和米糠对鲈鱼肝脏表现出肝损害所致的组织炎症反应、肝细胞水样变性和脂肪变性等病理特征.血粉、羽毛粉、双低菜粕组鲈鱼肠道皱襞出现顶端上皮细胞脱落、炎症细胞浸润等病理症状.