Effect And Mechanism Of Qing Gan Zi Shen Decoction On Heart Damage Induced By Obesity And Hypertension

Objective:Heart damage is a common target organ damage in hypertension,especially in patients with obesity and metabolic disorders.As an important transcription factor,Nrf2 plays a more and more important role in the regulation of heart structure and function.Qing Gan Zi Shen Decoction originated from the theory of blood stasis and heat of Professor Zhou Zhongying,a master of Chinese medicine in our hospital,but it is not clear whether the intervention effect and mechanism of Qing Gan Zi Shen Decoction on the target organ damage of obesity and hypertension.Therefore,the purpose of this study is to explore the intervention effect and potential mechanism of Qing Gan Zi Shen Decoction on the heart damage of obesity and hypertension.Methods:20 SHRs were randomly divided into four groups:standard diet group(STD),high fat diet group(HFD),low dose group(QL),high dose group(QH).Rats in the standard diet group were fed with standard diet,rats in the high-fat diet group and the treatment group were given HFD for 16 consecutive weeks,rats in the low-dose group and the high-dose group were given orally 1010mg/kg/day and 4040 mg/kg/day of water-soluble extract of Qing Gan Zi Shen Decoction for 16 consecutive weeks,and were tested as follows:(1)The blood pressure and body weight of SHR were measured,and the heart structure and function of SHR were detected by ultrasound;(2)HE staining and WGA fluorescence staining were used to detect the heart structure and the size of myocardial cells.The expression of ANP,BNP and ?-MHC in SHR heart tissue was detected by WB and IHC;(3)Masson and Sirius red staining were used to detect the degree of fibrosis in SHR heart tissue.The expression of TGF-?1,CTGF,collagen ?,collagen ? and fibronectin in SHR heart tissue was detected by WB and IHC;(4)The contents of TNF-? and IL-6 in serum and heart tissue were detected by ELISA,WB and IHC;(5)The mitochondrial morphology of heart tissue was detected by transmission electron microscopy.Nuclear protein,plasma protein and mitochondrial protein were extracted from heart tissue,and protein expression of CS,NRF-1,HO-1,SOD2,TFAM were detected by WB,IHC and IF;(6)The content of superoxide anion in heart tissue was detected by DHE staining.Protein expression and activity of 3-NT,4-HNE,8-OHdG,NQO1,GCLM,GCLC,GSR,GPX4,SOD1,CAT were detected by WB,IHC and related biochemical kits.The degree of protein carbonylation was detected using related kits and WB.using the content of MDA was detected using related biochemical kits.The contents of GSH and GSSG were detected by the related kits(7)Cytoplasmic protein,nuclear protein and mitochondrial protein were extracted and the expression of Nrf2 in heart tissue was detected by WB and IFResults:(1)Qing Gan Zi Shen Decoction can significantly reduce the body weight,systolic pressure,mean arterial pressure,LVID;d,LVID;s,LVPW;s and LV mass of HFD-fed SHR,and to a certain extent reduce the diastolic pressure,IVS;d,IVS;s and LVPW;d,and can significantly increase EF%,FS%and MV E/A of HFD-fed SHR.(2)Qing Gan Zi Shen Decoction can significantly improve the morphology of heart tissue,including making the arrangement of cardiac myocytes more orderly,reducing the degree of hypertrophy and swelling of cardiac myocytes,inhibiting the deposition of collagen fibers,reducing the surface area of cardiac myocytes.The potential mechanism is related to Qing Gan Zi Shen Decoction can significantly inhibit the expression of ANP,BNP and MYH7 in heart tissue.(3)Qing Gan Zi Shen Decoction can significantly inhibit collagen deposition and fiber proliferation in SHR heart tissue,and its potential mechanism is related to the inhibition of CTGF,TGF ?1,fibronectin,collagen ? and collagen ? protein expression in SHR heart tissue by Qing Gan Zi Shen Decoction.(4)Qing Gan Zi Shen Decoction can effectively inhibit the expression of TNF-?,IL-6 and PAI-1 in serum and heart,and reduce heart inflammation.(5)Qing Gan Zi Shen Decoction can effectively alleviate the damage of mitochondria such as swelling,number reduction,surface area and perimeter increase,crista disorder,crista decrease and mitochondrial cavity in the heart tissue of obese SHR,and can significantly increase the expression of CS.Its potential mechanism is related to the fact that Qing Gan Zi Shen Decoction can up-regulate HO-1,SOD2,TFAM and NRF-1 in the heart tissue of SHR.(6)Qing Gan Zi Shen Decoction can significantly reduce the content of ROS,4-HNE,MDA and 3-NT in heart tissue,inhibit the protein carbonylation and the expression of 8-OHdG,promote and increase the expression and activity of NQO1,GCLM,GCLC,GSTpi,GSR,GPx4,SOD1 and CAT,and increase the content of GSH and GSH/GSSG.(7)Qing Gan Zi Shen Decoction can significantly increase Nrf2 protein level in cytoplasm,nucleus and mitochondria of heart tissue.Conclusion:Qing Gan Zi Shen Decoction can effectively reduce weight and blood pressure,significantly improve the heart structure and function of obese SHR,inhibit myocardial hypertrophy,cardiac fibrosis,heart inflammation and oxidative stress,and improve the structure and function disorder of heart mitochondria caused by obesity and hypertension.Its potential mechanism may be that Qing Gan Zi Shen Decoction can increase the expression of Nrf2 in the cytoplasm and mitochondria,promote the nuclear and transcription of Nrf2.