RESUMEN
A 20-year-old woman with abetalipoproteinemia underwent orthotopic liver transplantation for cirrhosis, affording access to her liver and small intestine for study. Before transplantation, her plasma apolipoprotein B concentration was less than 1 mg/dL according to enzyme-linked immunosorbent assay, whereas after transplantation her plasma apolipoprotein B concentration was 76 mg/dL (all apolipoprotein B-100). Apolipoprotein B content was reduced in her intestine and liver compared with normal and cirrhotic controls. Cultured hepatocytes from the patient's explanted liver secreted a 1.006 g/mL less than or equal to d less than or equal to 1.063 g/mL lipoprotein rich in apolipoprotein E and a 1.063 g/mL less than or equal to d less than or equal to 1.21 g/mL lipoprotein containing apolipoproteins E and A-I with no immunodetectable apolipoprotein B in the culture medium. Normal hepatocytes secreted very low-density lipoprotein and low-density lipoprotein containing apolipoprotein B-100. Abetalipoproteinemic intestinal apolipoprotein B messenger RNA concentration was 4-5-fold higher than control values. However, the patient's liver apolipoprotein B messenger RNA level was one fifth that of control normal and cirrhotic liver. Analysis of the patient's intestinal and hepatic apolipoprotein B messenger RNA for posttranscriptional stop-codon insertion revealed normally edited transcripts. These results suggest that apolipoprotein B is synthesized as the product of a normally edited messenger RNA transcript, but not secreted, in abetalipoproteinemia.
Asunto(s)
Abetalipoproteinemia/genética , Apolipoproteínas B/biosíntesis , Expresión Génica , Intestino Delgado/metabolismo , Hígado/metabolismo , Abetalipoproteinemia/cirugía , Adulto , Apolipoproteínas/análisis , Apolipoproteínas B/análisis , Apolipoproteínas B/sangre , Secuencia de Bases , Colesterol/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Intestino Delgado/química , Hígado/química , Hígado/citología , Hígado/patología , Trasplante de Hígado , Datos de Secuencia Molecular , Oligonucleótidos , ARN Mensajero/análisis , Triglicéridos/sangreRESUMEN
To determine the effects of dietary and biliary lipid absorption on intestinal apo B-48 and apo A-I synthesis in the newborn piglet, 2-d-old female piglets were prepared with a duodenal infusion catheter. After recovery, animals were given either low triglyceride (Vivonex; VIV group) or high triglyceride (Intralipid; FAT group) diets by continuous intraduodenal infusion for 24 h. A bile-diverted group was also studied. Segments of proximal jejunum and distal ileum were then pulse-radiolabeled in vivo with 3H-leucine. Mucosal apo B-48 and apo A-I were immunoprecipitated, and apoprotein synthesis was expressed as percentage of total protein synthesis. Mucosal apoprotein content (ng apoprotein/microgram total protein) was measured by competitive ELISA assays. In jejunum and ileum, apo B-48 synthesis was not different in the three groups. However, apo B content increased 2.4-fold in jejunum and 1.7-fold in ileum in the FAT group compared with the VIV group. Immunoblotting revealed the majority of jejunal apo B to be apo B-48, not apo B-100 from contaminating plasma lipoproteins, in all three experimental groups. Bile-diverted animals had decreased jejunal apo B content compared with the VIV group. Jejunal apo A-I synthesis and content were approximately 2-fold higher in FAT animals compared with the VIV group. Although ileal apo A-I synthesis was also 2-fold higher in the FAT group, apo A-I content was not different from the VIV group. Neither jejunal nor ileal apo A-I synthesis was significantly affected by bile diversion, even though jejunal apo A-I content was decreased by over two thirds compared with the VIV animals. In the newborn piglet, intestinal synthesis of apo B-48 and apo A-I is differentially regulated by luminal lipid absorption. Although fat feeding and bile diversion regulate mucosal apo B-48 content, synthesis is unchanged, indicating a posttranslational regulatory mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Apolipoproteínas/biosíntesis , Mucosa Intestinal/metabolismo , Animales , Animales Recién Nacidos , Apolipoproteína A-I , Apolipoproteínas/metabolismo , Apolipoproteínas A/metabolismo , Apolipoproteínas B/metabolismo , Femenino , PorcinosRESUMEN
Fetal, newborn, and suckling piglets were used to study the intestinal expression of the apoA-IV gene in the immature mammal. Swine apoA-IV (42 kD) was isolated from fat-fed piglet lipoprotein-deficient plasma by adsorption to Intralipid followed by preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroelution. Rabbit anti-swine apoA-IV antibodies were raised, and apoA-IV was immunoprecipitated from small intestinal homogenates after in vivo radiolabeling with [3H]leucine. ApoA-IV synthesis was expressed as a percentage of total protein synthesis from trichloroacetic acid-precipitable counts. Fetal (40 day gestation) whole small intestine synthesis was 2.1%. Postnatally, 2-day-old newborn piglets given high triglyceride and low triglyceride duodenal infusions, as well as bile diversion, were studied. Synthesis rates in jejunal mucosa in all groups were comparable to the fetal whole intestinal value except in the jejunum of the high-triglyceride group, where synthesis was increased sevenfold. In 1- to 2-week-old fasting, cream-fed, and bile-diverted piglets synthesis was again unchanged except in the fat-fed jejunum, where synthesis doubled. Ileal synthesis rates in newborn and suckling animals were lower than jejunal rates and did not increase with lipid absorption or decrease with bile diversion. Northern blot hybridization of intestinal RNA samples from the newborn groups with an authentic cross-hybridizing human apoA-IV cDNA probe revealed a 1.8 kb signal which was strongest in the high-triglyceride jejunal samples. Slot blot hybridization showed eightfold increased apoA-IV mRNA levels in high-triglyceride jejunal samples as compared to low-triglyceride and bile-diverted jejunum with no differences in beta actin mRNA abundance.(ABSTRACT TRUNCATED AT 250 WORDS)