The Effect And Mechanism Of Gualou Xiebai Banxia Decoction On Heart Failure In Rats With Phlegm-Turbid Obstruction Syndrome

Objective:The research is to investigate the effect of Gualou Xiebai Banxia Decoction(GXBD)in im Proving heart failure in rats with Phlegm-turbid obstruction syndrome,and to study its pharmacodynamic molecular mechanism based on TGF-β1/Smad signaling pathway.Methods :1.SPF adult male SD rats were randomly divided into sham-operated blank control group,sham-o Perated original dose group,heart failure model group,the GXBD original dose treatment group,GXBD modern dose treatment grou and perindopril treatment group.After different treatments,the body weight and cardiac index of the rats were measured.The LVEF was measured by ultrasound,and the serum Renin,Ang II,ALD,TNF-α,IL-6,Cas Pase-3,Bax and TGF-β1 were measured by ELISA.Myocardial HE,Masson staining,TTC staining,immunohistochemistry and TUNEL staining were Performed.2.By using the Arraystar rat Lnc RNA V2.0 chi P,lnc RNA and m RNA high-throughput sequencing was performed on the myocardium of the rats in the sham-operated group,the model group and the GXBD treatment group.And bioinformatics analysis of the genes of interest were performed using GO and KEGG pathway analysis.According to ce RNA analysis and PCR verification,the network regulation map of lnc RNA-mi RNA-m RNA was constructed.3.SPF adult male SD rats were randomly divided into the sham operation group,the model group and the GXBD treatment group.After different interventions,q-PCR was used to detect myocardial TGF-β1,Col-Ⅰ,Col-Ⅲ m RNA expression and Western blot was used to detect the protein amount of TGF-β1,Col-Ⅰ,Col-Ⅲ,P-Smad2/3 and total Smad2/3.Results: 1.GXBD can obviously improve the cardiac function of rats with heart failure with phlegm-turbid obstruction syndrome,and the original dose group of this classic formula(referred to as classic formula)has the best effect.Compared with the sham-operated group,the rats in the heart failure model group lost weight,while the cardiac index increased significantly(P<0.01).Compared with the heart failure model group,the original dose of GXBD improved LVEF the most(P<0.05).Compared with the model group,each treatment group could reduce the serum concentrations of Renin,Ang II,ALD,TNF-α,IL-6,Cas Pase-3,Bax and TGF-β1,and the GXBD original dose group had the best effect(P<0.05).Compared with the model group,all treatment groups were improved.The GXBD original dose group had the most significant improvement in CVF(P<0.05),the most significant improvement in TTC staining necrosis area(P<0.05),and the smallest immunohistochemical score(P<0.05).The apoptosis index of TUNEL staining was the smallest(P<0.05).2.The differentially expressed rat myocardial lnc RNA and m RNA before and after treatment with GXBD were successfully screened.According to Fold change >2,P<0.05. compared with the sham operation group,the model group had 192 up-regulated lnc RNAs,357 down-regulated lnc RNAs,222 up-regulated m RNAs,and 456 down-regulated m RNAs.Com Pared with the model group,the GXBD treatment group had 1031 down-regulated lnc RNAs,430 up-regulated lnc RNAs,and 1042 m RNAs down-regulated 918 m RNAs raised.The genes up-regulated in the model group but down-regulated after GXBD treatment included 107 lnc RNAs and 115 m RNAs.After GO and KEGG pathway analysis of m RNA,it was found that GXBD may play a role in signaling pathways such as atherosclerosis,antigen processing and presentation,TGF-β,MAPK,c AMP,and hypertrophic cardiomyopathy.3.GXBD improved cardiac function in rats with phlegm-turbid obstruction syndrome by inhibiting the expression of TGF-β1/Smad signaling axis.Compared with the sham operation group,the m RNA expressions of TGF-β1,Col-Ⅰ,and Col-Ⅲ in the model group were significantly increased(P<0.05).Compared with the sham operation group,the protein levels of TGF-β1,Col-Ⅰ,Col-Ⅲ and P-Smad2/3 in the model group were significantly increased(P<0.05).All groups were significantly different(P<0.05).There were no significant changes in the total Smad2/3 protein content in the three groups.Conclusion: GXBD can improve cardiac function in rats with phlegm-turbid obstruction syndrome and the original dose of GXBD has the better effect.The molecular mechanism of improving heart failure may be to improve myocardial remodeling by inhibiting TGF-β1/Smad signaling pathway.