Identification and characterization of aquaporin-9 (AQP9) in porcine hepatic tissue and hepatocytes in monolayer culture     

Caperna TJ  Shannon AE  Richards MP  Garrett WM  Talbot NC 《Domestic animal endocrinology》2007,32(4):273-286

Aquaporins (AQPs) are members of a large family of integral membrane proteins involved in the rapid movement of water and neutral solutes across cell membranes. In this study, we have prepared an affinity-purified porcine-specific polyclonal antiserum to AQP9 and have investigated the distribution and expression of AQP9 in pig liver tissue and in hepatocytes in primary culture. Immunocytochemical analysis showed that AQP9 was primarily localized in the membrane structures of hepatocytes and was not associated with intrahepatic bile ducts or blood vessels. Western blot analysis indicated that AQP9 ranged in apparent molecular mass between 27 and 38 kD in whole liver and hepatocyte membrane fractions; minor components were also observed at approximately 34 kD in the cytosol compartment of hepatocytes, bile duct and gall bladder. A prominent immunoreactive band at 44 kD was shown to be an artifact of Western blot analysis. In primary cultures of porcine hepatocytes, glucagon enhanced absolute levels of AQP9 protein, while gene expression was enhanced by T3 and glucagon. Insulin alone had no discernable influence on AQP9 gene expression or its cellular protein levels. These data suggest that AQP9 is a major AQP in porcine hepatic tissue and appears to be primarily responsive to glucagon induction.  相似文献   

虹鳟胆管系统组织学观察   总被引：3，自引：1，他引：2  

黄布敏  袁莉刚  曲亚玲  谷来凤  朱俊峰 《甘肃农业大学学报》2013,48(1):31-36

采用常规苏木精.伊红(hematoxylin-eosin)H.E染色法、Masson三色染色法、Gomori银染法结合透射电镜研究技术,对虹鳟肝脏胆管形态结构进行了观察.结果表明:虹鳟肝脏被覆单层扁平上皮组织,肝实质内结缔组织较少,肝小叶结构不明显;汇管区结缔组织内含丰富的胶原纤维、网状纤维和弹性纤维;汇管区存在胆管孤管型、胆管动脉型、胆管静脉型和胆管动静脉型4种构型;肝细胞由双层肝板呈管式排列,双层肝板之间可见胆管上皮细胞;胆管、胆小管、前胆小管和胆毛细管构成肝脏胆管树系统.虹鳟肝脏胆管结构及多种汇管区构型特点表明虹鳟肝脏胆管微循环结构系统发育完善,肝脏的物质代谢能力旺盛.  相似文献   

Orexin-A stimulates the glucose output of porcine hepatocytes     

BIN Yan-fang SHU Gang ZHOU Li-hua CHEN Li-long ZHU Xiao-tong GAO Ping ZHANG Yong-liang ZHANG Shou-quan JIANG Qing-yan 《农业科学与技术》2009,(8):8-13

Orexin-A （OxA） is a key neuropeptide involved in the central control of appetite and the maintenance of energy homeostasis, as well as in the regulation of glycemia. In the present study, OxA receptors, OXtR and OX2R were identified to be highly expressed in porcine hepatocytes. 100 nM OxA could rapidly stimulate the glucose output of porcine hepatocytes in 10min. Primary hepatocytes treated by 1 nM-500 nM OxA for 24 h significantly increase the released glucose and the concentration of albumin, total bile acids and triglyceride in the supernatant. The mRNA expression level of such gluconeogenesis and fat mobilization related genes as acetyl-coA carboxylase, glycogen phosphorylase, fatty acid translocase and phosphoenolpyrurate carboxykinase were also up-regulated accordingly. The results first demonstrated that OxA had direct effect on the hepatic glucose and lipid metabolism, which was an aspect for OxA to exert its maintenance function of energy homeostasis.  相似文献   

多能干细胞来源的肝细胞诱导分化及其应用研究进展     

王洋  彭莹莹  段星祥 《西北农林科技大学学报(自然科学版)》2017,45(12):15-22

胚胎干细胞(embryonic stem cells,ESCs)和诱导多能干细胞(induced pluripotent stem cells,iPSCs)是一类具有多能性和自我更新能力的多能干细胞,这些细胞具有无限增殖能力,并有潜能分化为机体三胚层所有类型的细胞。多能干细胞来源的肝细胞能够作为肝细胞的替代物应用于体外药物筛选和肝脏疾病的治疗。文章对多能干细胞向肝细胞的诱导研究、多能性干细胞来源肝细胞的鉴定方案研究及其在药物筛选和疾病治疗方面的研究进行了综述分析,并对未来的研究方向进行了总结与展望。  相似文献   

胰岛素、胰高血糖素对体外培养犊牛肝细胞CPT-Ⅰ和CPT-ⅡmRNA表达的影响     

丁红研  李玉  史珍  杨威  张仁和  杨文涛  邓清华  刘国文  王哲  李小兵 《兽医大学学报》2014,(4):638-640

体外原代培养犊牛肝细胞,添加不同浓度的胰岛素（insulin,INS）（0、1、10、100、1 000nmol/L）和胰高血糖素（glucagon,GLN）（0、1、10、100、1 000nmol/L）,每个处理做3个重复,分别培养1h后,提取总RNA。实时荧光定量PCR（qRT-PCR）的方法,检测肉碱棕榈酰转移酶Ⅰ（carnitine palmitoyl transferase,CPT-Ⅰ）和肉碱棕榈酸转移酶Ⅱ（carnitine palmitoyl transferase,CPT-Ⅱ）mRNA表达的变化。结果显示,随着培养液中INS含量的升高,肝细胞中CPT-ⅠmRNA和CPT-ⅡmRNA的表达量逐渐降低,呈现一定的剂量依赖性。随着培养液中GLN含量的升高,肝细胞中CPT-ⅠmRNA和CPT-ⅡmRNA的表达量均升高,呈现一定的剂量依赖性,且当GLN的添加浓度高于100nmol/L时,CPT-ⅠmRNA和CPT-ⅡmRNA的表达量显著升高（P〈0.01）。结果表明,INS可在一定程度上抑制脂氧化关键酶的表达,GLN可显著促进脂氧化关键酶的表达。  相似文献   

枸杞多糖对四氯化碳诱导建鲤原代肝细胞损伤的保护作用研究     

刘英娟  杜金梁  贾睿  曹丽萍  王佳豪  殷国俊 《上海海洋大学学报》2014,23(5):718-725

研究中药枸杞多糖(Lycium barbarum polysaccharide,LBP)对四氯化碳(CCl4)诱导的建鲤(Cyprinus carpiovar Jian)肝细胞急性损伤的保护作用。利用8 mmol/L四氯化碳构建建鲤肝细胞损伤模型,用不同浓度的枸杞多糖处理肝细胞,设空白对照组、枸杞多糖对照组(0.4 mg/mL)、模型组(CCl4)、预防组(CCl4处理前加枸杞多糖孵育,0.1、0.2和0.4 mg/mL)、治疗组(CCl4处理后加枸杞多糖孵育,0.1、0.2和0.4 mg/mL)和预防治疗组(枸杞多糖孵育后经CCl4处理再经枸杞多糖孵育,0.1、0.2和0.4 mg/mL)。12 h后,收集细胞和细胞培养液,然后测定谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、细胞活力、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、免疫球蛋白M(IgM)及药物代谢酶细胞色素P450 2E1(CYP2E1)等生化指标。研究结果表明:CCl4处理后可以显著增加GPT、GOT、LDH、MDA、TNF-α、IL-1β、IgM及CYP2E1的含量,降低SOD活性和细胞活力;枸杞多糖处理组可不同程度地抑制上述生化指标的变化,其中预防组及预防治疗组对于转氨酶(GPT、GOT)、LDH、MDA、细胞因子(TNF-α、IL-1β)、IgM及CYP2E1等活性的升高皆有显著抑制作用,同时显著提高SOD酶活性及细胞活力,且呈剂量依赖性;而治疗组仅对LDH、TNF-α及CYP2E1有显著的抑制作用。综上结果,认为枸杞多糖对四氯化碳致建鲤肝损伤具有一定保护作用,可以应用于鱼类肝损伤的防治。  相似文献   

镉对大鼠原代培养肝细胞毒性的研究     

杨建泉  马宏兵  刘学忠 《现代农业科技》2012,(15):266-267,269

为探讨镉对原代大鼠肝细胞的毒性作用。用二步灌流法获得大鼠肝细胞,肝细胞暴露于浓度为2.5、5.0、10.0、20.0μmoL/L的醋酸镉中12、24 h,应用四甲基偶氮噻唑蓝(MTT)比色法检测醋酸镉对肝细胞存活率的影响及培养上清液中乳酸脱氢酶(LDH)的活性变化。结果表明,肝细胞暴露于浓度为2.5、5.0、10.0、20.0μmoL/L的醋酸镉中,细胞相对存活率显著下降(P0.01),LDH的释放量增加,且具有剂量效应关系。提示一定浓度的醋酸镉可引起肝细胞的毒性损伤。  相似文献   

Effect of propionate, pyruvate and β-hydroxybutyric acid on pyruvate carboxylase mRNA expression of in vitro culture bovine hepatocytes     

Chuang XU  Zhe WANG 《Animal Science Journal》2006,77(4):440-446

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大口黑鲈原代肝细胞的培养及其应用于CYP450活性的诱导     

喻文娟  李聃  王翔凌  张宁  邱军强  杨先乐 《海洋渔业》2008,30(1):31-36

体外培养大口黑鲈(Micropterus salmoides)原代肝细胞,观察其在培养过程中的形态变化,同时检测培养上清液中的乳酸脱氢酶(LDH)、白蛋白(ALb)含量,从而判定肝细胞的增殖和代谢能力,进而应用于CYP450的诱导研究。结果表明:培养5 d后,肝细胞连接成片,分裂增多;培养至7 d,细胞渐至单层;14 d后,细胞停止生长,出现老化迹象。培养至第3~4天的细胞上清液中LDH的渗出量呈上升趋势,在第4天时达到峰值,随着培养时间的延长,肝细胞的酶渗出量逐渐减少。ALb的分泌量在第3~5天呈上升的趋势,第4~8天维持在较高水平,在第5天达到最高,此后随着培养时间的延长,ALb的分泌量呈下降趋势,故选择培养至第5天的大口黑鲈肝细胞应用于CYP450酶系中的氨基比林-N-脱甲基酶(ANDM)的诱导研究。40μM利福平诱导24 h后,ANDM活性最高。ANDM的诱导研究为鱼类细胞药物代谢酶体外诱导奠定基础。  相似文献   

Distribution of apolipoproteins A-I and A-II in serum, secreted proteins by cultured hepatocytes, and the plasma membrane of hepatocytes in eel     

Hironori Kato  Ying-Ping Ge  Djibril Ndiaye and  Seiichi Hayashi 《Fisheries Science》2000,66(3):579-585

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