Experimental Study On The Effect And Mechanism Of Qing Gan Zi Shen Tang In Improving Hypertension Complicated With Metabolic Disorder By Upregulating SIRT1

Background and ObjectiveEssential hypertension is usually associated with metabolic disorders,including obesity,dyslipidemia,glucose intolerance and insulin resistance.Qing Gan Zi Shen Tang(QGZST)is a famous traditional Chinese medicine formula in the Jiangsu Province Hospital of Traditional Chinese Medicine for its efficacy in treating hypertension,obesity,hyperlipidemia and insulin resistance.It has the function of clearing away liver-fire and nourishing yin and tonifying the kidney.In previous studies,we found that it can be proved to be able to improve the clinical symptoms of hypertension and metabolic disorders.QGZST plays an important role in regulating lipid metabolism in SHR rats fed with high fat diet,and can significantly improve lipid deposition in the liver of SHR rats.However,The effect and molecular mechanism of QGZST on SIRT1 in SHR rats fed by high-fat diet have not been studied.Therefore,We used High-fat feeding SHR rat model to explore the effect of Chinese herbal medicine on SIRT1 and its possible mechanism,which provides a theoretical basis for the treatment of hypertension complicated with metabolic disorder with traditional Chinese medicine.Method(1)Fifteen 4-week-old spontaneously hypertensive SHRs were randomly divided into three groups:normal group(STD),high fat diet group(HFD)and QGZST group(5 rats in each group).STD group was fed with normal diet,HFD group and the QGZST group were fed with high fat diet.After 4 weeks of high fat feeding,the QGZST group was given suspension of QGZST,and STD group and HFD group were given the same volume of distilled water for 9 weeks,once a day.The average daily dietary intake was measured and weighed weekly.After 4 weeks of gastric perfusion,the blood pressure were measured.After 9 weeks of gastric perfusion,the blood pressure were measured.Each rat were collected 2 ml of centrifugal blood for preservation.After anesthesia,all the rats were killed,and the epididymal fat was taken,then embedded in paraffin for preservation.To investigate the effects of QGZST aon body weight,food intake and blood pressure of spontaneously hypertensive rats fed with high fat.(2)Serum lipids were measured by enzyme-linked immunosorbent assay(Elisa),the size of epididymal adipocytes was observed by hematoxylin and eosin(HE)staining,and the protein expression of SIRT1,C/EBPa,PPARy,FABP4 in epididymal adipose tissue were detected by Western blot.Reverse transcription polymerase chain reaction by reverse transcriptase polymerase chain reaction(RT-PCR)for detection of the expression of mRNA in SIRT1,C/EBPa,PPARy,FABP4,which was studied to investigate the effect and mechanism of QGZST on blood lipids and adipocytes in spontaneously hypertensive rats fed with high fat.(3)The levels of inflammatory factors and fat factors in serum of rats were detected by Elisa.The expression of epiddymal adipose tissue inflammatory factors(TNF-?,IL-6,Ang ?)and adiponectin(leptin,adiponectin)were detected by reverse transcriptase polymerase chain reaction(RT-PCR).Determination of FOXO1,PTP1B,NF-KB-p65 and acetyl-NF-KB-p65(K310)were detected by Western blot.To investigate the effect and mechanism of QGZST on inflammatory and fatty factors in spontaneously hypertensive rats fed with high fat.Results(1)QGZST can reduce body weight,blood pressure,especially systolic blood pressure(SBP)and mean arterial pressure(map)of high fat fed SHR rats.(2)QGZST can reduce the fat tissue of SHR rats fed by high fat,diminish the fat cells and reduce the blood lipid level.(3)QGZST can reduce the expression of SIRT1 protein and mRNA in high fat fed SHR rats,and increase the expression of C/EBP?,PPAR?,FABP4 protein and mRNA.(4)QGZST can reduce the level of TNF-?,IL-6,Ang ? and leptin,decrease the expression of mRNA and increase the expression of adiponectin and its mRNA in SHR rats fed with high fat.(5)QGZST can decrease the expression level of inflammatory factors,adipose factor-related signaling molecules acetyl-NF-KB-p65 and PTP1B in hyperlipidemic SHR rats and increase the expression of FOXO1 protein.ConclusionHigh fat diet inhibited SIRT1 and FOXO1 and activated C/EBPa,PPAR-y FABP4,acetyl-NF-Bp65 and PTP1B,which resulted in dysfunction of adipose tissue..By up-regulating SIRT1 and FOXO1 and down-regulating C/EBPa,PPAR-y FABP4,acetyl-NF-Bp65 and PTPIB,QGZST can raise SIRT1 to improve the microenvironment of adipose tissue,and effectively intervene hypertension and metabolic disorder.