The Study Of Relation Of Phospholipase C And Essential Hypertension

Objective: To study the content change of phospholipase C-γ1 ( PLC-γ1) and phospholipase C -δ1(PLC-δ1) in essential hypertension,and make clear their role in the disease.Methods: 30 patients were divided into two groups complying with their medical record :with(N=13) or without(N=17) essential hypertension.Their cholecyst artery Vascular smooth mucscle cells(VSMCs) were homogenated and centrifuged ,then lysate were abstracted and denatured by adding equal sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) buffer and boiling for 5 minutes. lysate protein concentration was determined in duplicate foreach sample by the method of Bradford .equal amounts of protein(10μg/lane) were resolved on an 8% SDS- polyacrylamide gel,eletrophoresed at 100v for 1.5h,and transferred to Polyvinylidene difluoride(PVDF) membranes at 4℃ and 70mA for 14h.Bands were blocked in 5% dried milk for 1h at room temperature(23-25℃) and probed with primary antibody which is anti-PLC-γ1 monoclonal antibody of bovine or anti-PLC-δ1 monoclonal antibody of human for 1h at room temperature, and with horseradish peroxidase(HPR)-conjugated secondary antibody for 1h at room temperature.Band intensity was detected by enhanced chemiluminescenece(ECL) and exposure to Kodak X-ray film which was scanned by laser scanner.Band's value of integral absorbency was analysed by ImageMarker Totallab software ,and it was the relative content of PLC-γ1 or PLC-δ1 in VSMCs .Results: Both PLC-γ1 and PLC-δ1 in VSMCs can be detected,but there was no significant difference between the essential hypertension group and normotension group(t=1.101,0.682,p>0.05).Conclusions: 1)Both PLC-γ1 and PLC-δ1 are exist in VSMCs,and they all play a role in the signal transduction of VSMCs. 2)The content of PLC-γ1 or PLC-δ1 is no change,which is probable that there is no relationbetween the change of PLC activity in EH and the content of PLC-γ1 and PLC-δ1.3)It is an effective way to quanlify slight trace protein with western blotting,exposing to X-ray film,scanning with laser scanner and analyzing integral absorbency with Totallab software which replaces the content of protein.