Effect Of Ginkgo Biloba Extract On OSAHS With Hypertension Rats Adipocytes By TLR4/JNK Signaling Pathway

Objective:OSAHS is the major causes of secondary hypertension.Disorder of sympathetic nerve disorder and RAS system could injury vascular endothelial cells and heighten the level of inflammatory reaction after repeated hypoxia and low ventilation.Some support that obesity and hypertension are chronic inflammatory diseases,and inflammation plays an important role,so we guess that inhibiting inflammation could reduce the level of hypertension of OSAHS.Past studies has confirmed Ginkgo biloba extract?GBE?could reduce the level of hypertension of OSAHS model rats.We established the model of OSAHS with hypertension adipocytes to observe the mechanism of GBE on TLR4 dependent JNK pathway.Methods:The experiment consisted of three steps.The first stage:Rats were randomly divided into normal oxygen control group?NC group?,intermittent hypoxia group?IH group?and the GBE therapy group?GBE group?.GBE group were irrigated with 1mL/kg GBE per day,NC group and IH group were irrigated with saline water.IH group and GBE group were given intermittent hypoxia environment 8 hours every day.The change of tail blood pressure and physiological activities were observed.Using normal^[1]and improved digestion method to culture preadipocytes.The morphology of cells were observed under inverted microscope,the proliferation were analyzed by flow cytometry,identified by Oil red O staining.MTT method was used to test the influence of different concentrations and time of GBE.The second stage:the model cells were divided into IH group,IH+LPS group,IH+TAK242+LPS group and IH+SP600125+LPS group,the expression of mRNA and protein about TLR4 and JNK were measured by PCR and Western Blot,and the level of TNF-a,IL-1?and IL-6 were tested by ELISA.The third stage:model adipocytes were divided into IH group,IH+LPS group,IH+GBE group,IH+GBE+LPS group.The expression of TLR4 and JNK mRNA and protein were tested by PCR and Western blot,and the level of TNF-a,IL-1?and IL-6 were tested by ELISA.Results:The first part:?1?Rats in model group turned to drowsiness,activity reduced,nasal lip cyanosis,and breath like purr,and blood pressure increased?P<0.05?.But the GBE group didn't had significant changes.?2?After 12 hours some round cells were adherent,turned to long spindle for 24 hours,3 or 5 days later growth into logarithmic phase,cytomixis after 8days.The cells stopped growth with lipid inducing solution,transparent refractile granules turned to droplets which could be dyed into orange with Oil Red O staining,nucleus were pushed to other side.?3?Flow cytometry assay showed cells had the ability to divide,and the promoted method group were higher.?4?GBE could promote the growth of preadipocytes,and 100mg/L,12hours was the best situation.The part two:The expression of TLR4 and JNKmRNA in IH+LPS group were higher compared with other groups?P<0.05?,Western Blot and ELISA results also true.The third part:The level of TLR4 and JNK mRNA in IH+GBE group were lower than other groups?P<0.05?.Results of protein and ELISA were same.Conclusions:?1?Changed the extraction digestion conditions could improve the ability of proliferation and division of preadipocytes.?2?The GBE had time and concentration dependence on the growth of preadipocytes.?3?TLR4 mediated JNK signal pathway plays a role in the development of OSAHS with hypertension.?5?The mechanism of that GBE reduces hypertension caused by OSAHS maybe is inhibiting the TLR4 mediated JNK pathway of adipocytes,to reduce the expression of inflammation.