纳米氧化铜对鸡肝细胞CP mRNA表达及培养基质中肝酶活性的影响

将培养48h的鸡原代肝细胞随机分为13组,每组3个重复。第1组为对照组,第2～5、6～9和10～13组分别以硫酸铜、氧化铜和纳米氧化铜的形式添加铜2、4、8、16mg/L,继续培养24h后检测上清中AKP、AST、ALT、γ-GT和LDH活性,用实时荧光定量PCR法检测肝细胞CP mRNA表达量。结果显示,以硫酸铜形式添加铜2～16mg/L、以氧化铜形式添加铜8～16mg/L和以纳米氧化铜形式添加铜16mg/L,培养基质中γ-GT活性均显著高于对照组（P〈0.05）;添加铜2mg/L时,硫酸铜组γ-GT活性显著高于纳米氧化铜组（P〈0.05）,添加铜8mg/L时,氧化铜组γ-GT活性显著高于纳米氧化铜组（P〈0.05）。添加铜2mg/L时,硫酸铜和氧化铜组AST和LDH活性显著高于对照组和纳米氧化铜组（P〈0.05）。添加铜2mg/L时,氧化铜组ALT活性显著高于纳米氧化铜组（P〈0.05）。添加铜8mg/L时,硫酸铜和氧化铜组AKP活性显著高于对照组和纳米氧化铜组（P〈0.05）。添加铜2～8mg/L时,肝细胞CP mRNA表达量显著高于对照组（P〈0.05）;添加铜2mg/L时,纳米氧化铜组肝细胞CP mR-NA表达量显著高于氧化铜组（P〈0.05）,氧化铜组显著高于硫酸铜组（P〈0.05）;肝细胞CP mRNA表达量随铜添加量的增加而降低。结果表明,基质中铜达到2mg/L时,纳米氧化铜比硫酸铜、氧化铜更有利于肝细胞生长,纳米氧化铜对体外培养肝细胞的损害作用低于硫酸铜和氧化铜。     

Changes in collagen fiber content and hepatic stellate cell distribution in the liver of chick embryos and growing chickens

The content of collagen and the distribution of hepatic stellate cells (HSCs) were studied to elucidate the occurrence of sex‐dependent variations in the liver of developing embryos and growing chickens. Chick embryos from embryonic days (e) 12 to e20 and chicks at 1, 4 and 8 weeks were analyzed. Liver tissue was processed using NaOH maceration and freeze‐dried to obtain the collagen fiber specimens. HSCs were identified by double fluorescent immunohistochemistry for desmin and vimentin. There were no sex‐dependent variations in the percentage of collagen fiber per liver weight and HSC area during embryonic stages. However, the content of collagen fiber increased during embryonic development in both sexes. On the other hand, the area of HSCs significantly increased in growing males but did not show any change in females. Importantly, sex differences were observed in both collagen fiber content and HSC area in the liver at 8 weeks. These results indicate that the occurrence of collagen content variations takes place at 8 weeks in chicken liver, suggesting that a sex‐dependent hormone may play an important role on the collagen production of HSCs in the growing chicken liver.     

HELICAL COMPUTED TOMOGRAPHIC ANGIOGRAPHY OF CANINE PORTOSYSTEMIC SHUNTS

Helical computed tomographic (CT) angiography was performed in 16 dogs with known or suspected portosystemic shunts. Fifteen portosystemic shunts were detected including five single intrahepatic shunts, five single extrahepatic shunts, and five multiple extrahepatic shunts. One dog had a normal CT examination. All diagnoses were confirmed by one or several alternate methods including ultrasound, surgery, necropsy, angiography, and liver biopsy. CT detected the origin of 13 of 15 portosystemic shunts and insertion of 13 of 15 shunts. Limitations included inability to resolve two vessels originating very close to each other, and identification of vessels that traveled parallel to the axial image plane. CT angiography is a promising, minimally invasive method of diagnosing a variety of portosystemic shunts in dogs.     

THREE-DIMENSIONAL HELICAL COMPUTED TOMOGRAPHIC ANGIOGRAPHY OF THE LIVER IN FIVE DOGS

The objective of this study was to develop a simple, safe, minimally invasive protocol to evaluate the hepatic vasculature. Five purpose-bred Beagle dogs underwent noncontrast-enhanced computed tomographic scan of the entire abdomen. A dynamic, nonincremental computed tomography scan at the level of T11 was then performed using a test bolus of contrast medium to determine time to peak opacification and to aid in the calculation of scan delay. The time to peak arterial enhancement ranged from 2.0 to 7.0 s, with a median of 2.0 s. The time to peak portal venous enhancement ranged from 23.0 to 46.0 s, with a median of 32.0 s. Scan delay for arterial opacification ranged from 0 to 5.0 s, with a median of 0 s. Scan delay for the portal phase of opacification ranged from 6.0 to 21.0 s, with a median of 17.0 s. Using this information, two separate computed tomographic studies were used to image the arterial and portal venous phases of circulatory opacification, respectively. The dogs were hyperventilated to prevent breathing motion during the scan, each of which required approximately 20 s. A power injector was used to inject diatrizoate meglumine (128 mg I/kg) through an 18-gauge cephalic vein catheter at a rate of 5 ml/s. Scanning was initiated after the appropriate scan delay to optimize the specific phase of vascular filling. Maximum intensity projections allowed clear delineation of the hepatic arteries and the portal venous system, while eliminating overlying structures that might interfere with image analysis. Time/density curves were generated, and the time needed for each study was recorded. Hepatic arteries and portal veins were clearly visualized in all dogs. Parenchymal opacification was also observed.     

VALIDATION OF DECONVOLUTIONAL ANALYSIS FOR THE MEASUREMENT OF HEPATIC FUNCTION IN DOGS WITH TOXIC-INDUCED LIVER DISEASE

The extraction of the hepatobiliary radiopharmaceutical ^99mTc-mebrofenin ( Choletec ) by the liver can be used to evaluate the severity of hepatocellular disease. The hepatic parenchymal cells extract mebrofenin from the blood by the same active transport mechanism as bilirubin. The ability of the liver to extract ^99mTc-membrofenin is a measure of hepatic parenchymal cell function. In this study, we induced hepatocellular disease by administration of a hepatotoxic drug and compared a direct method of determining the hepatic extraction of ^99mTc-membrofenin to hepatic extraction fraction derived from deconvolutional analysis. We also compared both methods of calculating the hepatic extraction of ^99mTc-membrofenin to liver histopathology. Hepatic extraction fraction derived from deconvolutional analysis correlated very well to the direct measurement technique (R=0.922, p<0.001). Both methods of determining hepatic extraction correlated well to quantitative histopathology, having the same correlation coefficient and p values. (R=-0.833, p=0.003). As the hepatic extraction ^99mTc-membrofenin decreased, the severity of the histopathologic lesions of the liver increased in a linear fashion. There was a significant correlation of the hepatic excretion T_1/2 to quantititative histopathology (R=0.949, p<0.001). The hepatic excretion T_1/2 increased as the severity of the histopathologic lesions of the liver increased. Hepatic extraction (HEF) and excretion of ^99mTc-membrofenin are good predictors of the severity of hepatocellular damage in toxic induced liver disease. This study helps validate the premise that HEF derived from deconvolutional analysis ois a good predictor of the actual first pass hepatic extraction of ^99mTc-membrofenin.     

茯苓配方浸膏对高脂饮食小鼠肥胖及脂质沉积的影响

为了探究茯苓配方（茯苓、山楂、黄大茶）浸膏对高脂饮食小鼠肥胖及脂质沉积的影响,将C57/BL6N小鼠随机分为对照组,模型组,奥利司他药物对照组,茯苓配方（Poria cocos Wolf formula,PWF）浸膏低、高剂量组。除对照组给予低脂饲料,其他组均以高脂饲料喂养12周。每周记录小鼠的体重、能量摄入。给药结束后,计算各组小鼠Lee’s指数、脏器系数,采用苏木精-伊红（hematoxylin-eosin,HE）染色法观察各组小鼠肝细胞与脂肪细胞形态变化,全自动生化仪测定各组小鼠血清中TG、TC、HDL-C、LDL-C、AST及ALT水平变化。结果显示,与对照组相比,模型组小鼠体重、Lee’s指数、脏器系数、TG、TC、LDL-C、AST及ALT水平均显著升高（P<0.01或P<0.001）,肝脏脂滴数量与脂肪细胞直径增加;与模型组相比,茯苓配方浸膏高剂量组小鼠体重、Lee’s指数和脏器系数均显著降低（P<0.05或P<0.01）,低剂量组小鼠体重、Lee’s指数和脏器系数均没有显著变化,茯苓配方浸膏高剂量组小鼠肝脏脂滴数量与脂肪细胞直径减少,TG、TC、...     

Curcumin attenuates lipopolysaccharide/D-galactosamine-induced acute rat liver injury via activating autophagy of hepatocytes

AIMTo investigate the effect of curcumin (CUR) on autophagy of hepatovyte in rats with lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced acute liver injury (AHI). METHODSThe healthy male Sprague-Dawley (SD) rats were randomized into the control group,AHI group,CUR group, 3-methy-ladenine (3-MA) group and 3-MA+CUR group, with 6 rats in each group. AHI was induced with an intraperitoneal injection of LPS and D-GalN. Liver function was tested 12 h after LPS/D-GalN treatment. Pathological changes of liver tissues were analyzed by HE staining.The amount of autophagic bodies were observed by transmission electron microscopy. The protein levels of autophagy related-proteins LC3 and beclin 1 in livers were detected by Western blot. ELISA were used to examine the serum levels of tumor necrosis factor-α (TNF-α). RESULTSCompared with control group, the serum level of alanine aminotransferase (ALT) and asparated aminotransferase (AST) were significantly increased, hepatic pathological damage were aggravated and serum TNF-α level was significantly increased in AHI group, while the autophagic bodies and the protein levels of LC3 and beclin 1 were increased (P<0.01). Compared with AHI group, the serum level of ALT and AST were significantly decreased, hepatic pathological damage were attenuated and serum TNF-α level was significantly reduced (P?<0.05), while the autophagic bodies and the protein levels of LC3 and beclin 1 were significantly increased in CUR group (P<0.01). Compared with CUR group, the serum level of ALT and AST were significantly increased, hepatic pathological damage were aggravated and serum level of TNF-α was significantly increased in 3-MA group and 3-MA+CUR group, while the autophagic bodies and the protein levels of LC3 and beclin 1 were decreased (P<0.01). CONCLUSION Curcumin protects rats against LPS/D-GalN-induced liver injury, partially due to activation of hepatocyte autophagy in livers.     

胎儿肝干细胞的分离培养与鉴定

从人胎儿肝脏中分离、纯化肝干细胞并进行鉴定。采用改进Seglen胶原酶原位灌注消化,结合Percoll密度梯度离心分离纯化胎儿肝干细胞,计数并测定细胞活性,光镜、电镜对其形态进行观察,用免疫细胞化学法(ICC)检测细胞表面标志表达情况。结果表明,所分离细胞活性率为90.80%±2.04%,细胞平均为(2.74±0.77)×107个/肝(n=10),光镜、电镜下观察其具有肝干细胞形态及超微结构特点,ICC染色,AFP、CK19、CK8及CD34等呈阳性,原代培养可形成肝干细胞集落。研究结果为人胎儿肝干细胞的存在提供了直接依据。