Effect Of17-β Estradiol Exposure On Connective Tissue Growth Factor Expression In Cultured Mice Vascular Smooth Muscle Cells By High Glucose

Chapter one High glucose increases the expression of CTGF in cultured mice vascular smooth muscle cellsObjective:This Part,we investigated the effects of high glucose on the expression of connective tissue growth factor (CTGF) in cultured mice vascular smooth muscle cells (mVSMCs).Methods:mVSMCs were divided into three groups and cultured in three different culture conditions for0、6、12、24、48hours or0、12、24、48、72hours, respectively:normal glucose group (NG group,5.5mm-ol/L Glu), mannitol group (5.5mmol/L Glu+19.5mmol/L mannitol), high glucose group (HG group,25mmol/L Glu), Real-Time PCR and Western blot were used to detect the changes in CTGF mRNA and protein expressions in the cells after the treatment.Results:The expressions of CTGF mRNA were significantly higher in HG group than in NG group (P<0.05). After high glucose treatment, the CTGF mRNA expression increased gradually and reached the peak level at6h, then followed by gradual decrease till recovering the baseline level at48h. However, mannitol group did not differ significantly from NG group in the expressions of CTGF mRNA.The expression level of CTGF protein also increased progressively and reached the peak level at24h, recovering the base-line level at72h.Conclusion:High glucose increases the expression of CTGF in mice vascular smooth muscle cells. Chapter Two Effect of17β estradiol exposure on connective tissue growth factor expression in cultured mice vascular smooth muscle cells by high glucoseObjective:In the above study, we first found high glucose increasesd the expression of CTGF in mice vascular smooth muscle cells. In the present study, we investigated the effect of17(3estradiol exposure on connective tissue growth factor expression in cultured mice vascular smooth muscle cells by high gluclose.Methods:After being cultured and passaged, the VSMCs were cultured in six different culture conditions for12hours or24hours, respectively:normal glucose group (NG group,5.5mmol/L Glu), high glucose group (HG group,25mmol/L Glu) plused different concentrations of17(3estradiol group (End concentration were OnM、0.01nM、1nM、100nM、1000nM), then the expression of CTGF were detected by Real-Time PCR technique and Western blotResults:Real-Time PCR results showed that compared with the normal control group, high glucose groups including different concentrations of17(3estradiol promoted the expression of mVSMCs CTGF mRNA (P<0.05); And compared with group without17(3estradiol,0.01nM、1nM、100nM、1000nM17(3estradiol groups had no significant difference on expression of CTGF.Western Blot results showed that:compared with the normal control group, and the expression level of CTGF were increased in the high glucose groups containing different concentrations of17(3estradiol groups; the protein expression of CTGF were induced by0.01、1、100、1000nM17(3estradiol group compared with without17β estradiol but there was no differences between each group.Conclusion:17β estradiol promote CTGF protein expression induced by high glucose, but not on CTGF mRNA demonstrating that17(3estradiol may play a certain role in the process of diabetes related atherosclerosis.