| [Objective]Familial hypercholesterolemia (FH), which is the most severe disorder of lipid metabolism, is one kind of monogenic autosome dominant inherited diseases. It is characterized by high levels of cholesterol, which frequently gives rise to tendon xanthenes and premature coronary artery disease. It has been widely accepted that low density lipoprotein receptor(LDL-R) mutation causes FH. Recently, some FH subtypes have been identified and the clinical manifestation is very similar to that of FH. To investigate LDL-R function of FH patients and the possible virulence gene of FH and to approach the relationship between the binding and internalization function of the LDL-R on the FH patients' lymphocytes and their phenotypes, we find a clinical definite FH patient and exam the lipid level of the family. And we build a method for immortalizing lymphocytes and analyze the expression and the binding and internalization function of LDL-R of proband and proband's parents. To address the relationship of gene mutation and clinical manifestation, we screen the Apo BlOO, LDL-R and adaptor protein gene mutations.[Methods]We studied a FH proband, which was diagnosed by clinical features and blood lipid tests. The proband and proband's parents were also applied with cardiovascular ultrasound. Lymphocytes were isolated from the whole blood of the health adult, proband and proband's parents and immortalized. The lymphocytes were cultured in 1640 medium containing 10% human lipoprotein-deficient serum (LPDS) for 72 hours to induce LDL receptor's activity. Then the lymphocytes were incubated with fluorescein isothiocyanate (FITC) labeled LDL at 4℃ and 37℃ for 2 hours respectively. The binding and internalization of LDL to LDL receptor were detected by flow cytometer (FCM) . Also we analyze gene and LDL-R function using PCR, FCM and immunohistochemistry staining. Apo B100 point mutation was detected by PCR... |
