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Objective To investigate the regulatory effect of Rab1 on the expression of β-adrenergic receptors (β-ARs) in rat pulmonary microvascular endothelial cells (RPMVECs). Methods Primary cultured RPMVECs served as the study object. Three experiments were designed to detect: 1) the effect of sea water on β-ARs expression, for which a control group and sea-water treatment group were set up; 2) the effect of Rab1 WT (wild-type rat lentivirus Rab1 expression vector) and Rab1 N124 I transfer on the expressions of β-ARs and its subtypes β1-AR and β2-AR, for which Rab1 WT and Rab1 N124 I were established, and then a control group, Rab1 WT group and Rab1 N124 I group were set up; 3) the effect of sea water on the β-ARs expression in the Rab1WT-transferred RPMVECs, for which a control group, sea-water treatment (SW) group and WT/SW group (sea water treatment after Rab1WT transfection) were set up. The expressions of β-ARs and its subtypes (β1-AR and β2-AR) in RPMVECs were determined by ligand binding assay. Results 1) In the sea water experiment, the cell-surface expressions of total β-ARs were 1324.00±130.01 and 2107.00±202.36cpm/105 cells, respectively, and the difference between them was statistically significant. 2) In the experiment, the expression levels of β-ARs were 2048.00±207.64, 2833.00±203.07 and 1040.00±91.59cpm/105 cells, respectively, in control group, Rab1 WT group and Rab1 N124I group, and the expression levels of β1-AR in the aforementioned three groups were 517.78±29.65, 591.22±18.29 and 364.11±20.68cpm/105 cells, respectively, and those of total β2-AR were 1627.90±112.27, 2262.10±169.33 and 694.33±29.79cpm/105 cells, respectively. After Rab1 WT transfer, the expression of β1-AR in RPMVECs increased by 14.2% (P > 0.05), and that of β2-AR by 49.0% (P < 0.01), as compared with the control group. 3) In the experiment, the expression level of total β-ARs was 1276.00±121.42cpm/105 cells in SW group, decreased by 38.0% as compared that of the control group (2059.00±174.62cpm/105 cells, P < 0.01), and 2017.20±101.14cpm/105 cells in WT/SW group, showing no statistically significant difference as compared with control group (P > 0.05), but increased by 58% as compared with SW group (P < 0.01). Conclusion Sea water may decrease the β-ARs expression on the membrane of RPMVEC, and Rab1 seems to have an antagonistic effect, which could up-regulate the expression of β-ARs on the membrane of RPMVEC. |
