| Objective: 1.To explore the clinical significance of Gal-3 and inflammation in chronic heart failure through clinical cross-sectional studies.2.Evaluate the clinical efficacy of Luqi Fang on patients with chronic heart failure with Yang deficiency and blood stasis syndrome through clinical randomized controlled studies,and preliminarily explore its mechanism.3.Observe the effects of Luqi Decoction on Gal-3 and NLRP3/IL-1βsignaling pathways through animal experiments,explore its mechanism of inhibiting inflammation and improving myocardial fibrosis,and provide a theoretical basis for Luqi Decoction in the treatment of chronic heart failure.Methods:Cross-sectional study: 138 patients who met the diagnostic criteria for chronic heart failure and 62 healthy subjects were collected.Observation;(1)Comparison of LVESD,LVEDD and LVEF indicators of cardiac function between the two groups;(2)Sera from patients with different cardiac function classifications Comparison of Gal-3,hs-CRP,TNF-α and IL-6 levels;(3)The correlation between serum Gal-3,hs-CRP,TNF-α and IL-6 levels and MACE;(4)Analysis of Gal-3 subjects Working curve.Randomized controlled study:Collect 90 patients who meet the diagnostic criteria of chronic heart failure with Yang deficiency and blood stasis syndrome,and randomly divide them into a control group and a treatment group,with 45 cases in each group.The control group was given standardized treatment with Western medicine,and the treatment group was treated with Luqi Fang on the basis of standardized treatment with Western medicine for 12 weeks.Observation before and after treatment:(1)Serum Gal-3,NLRP3,PCP,PIIINP levels;(2)Cardiac ultrasound: LVEF,LVEDD,LVESD,LVPWT,IVS,LVMI,LVM,LVEDVI,LVESVI indicators;(3)6-minute walk test;(4)Minnesota quality of life Score;(5)changes in cardiac function(NYHA classification);(6)TCM syndrome score.Animal experiment: After successfully making the heart failure model of 60 Wistar rats after myocardial infarction,they were randomly divided into six groups: sham operation group,model group,model+LQF-L group,model+LQF-M group,model+LQF-H Group,model+MCP group,10 animals in each group.The sham operation group and the model group were given an equal volume of normal saline for intragastric administration,the LQF-L group was given a low dose of Luqifang 0.625g/(kg·d),and the LQF-M group was given a medium dose of Luqifang 1.250g/(kg ·D),LQF-H group 2.500g/(kg·d),1m L/kg gavage,once a day.MCP group was given modified citrus pectin 1mg/(kg·d),1m L/kg gavage,once a day;after drug intervention for 4 weeks,observe:(1)The changes in body weight and activity of rats in each group;(2)HE and Masson staining detection Pathological changes of the myocardial tissues of rats in each group;(3)Echocardiography to observe the improvement of cardiac function of rats in each group;(4)Western blot to detect the protein levels of Gal-3,NLRP3,Capase-1,IL-1β,PICP,PIIINP;(5)RT-PCR Detect the expression changes of Gal-3,NLRP3,Capase-1,IL-1βm RNA;(6)Immunohistochemical observation of the expression of Gal-3,PICP,PIIINP in myocardial tissue;(7)ELISA to detect serum TNF-α,IL-Iβ in each group of rats,IL-18 content level;Results: Clinical cross-sectional study:(1)Compared with the healthy control group,the serum Gal-3,hs-CRP,TNF-α and IL-6 levels of patients in the heart failure group were significantly increased(P<0.01),and the difference was statistically significant(2)The levels of serum Gal-3,hs-CRP,TNF-α and IL-6 in heart failure patients with heart function grade IV are higher than those with heart function grade III heart failure,and serum Gal-3 in patients with heart function grade III heart failure,Hs-CRP,TNF-α and IL-6 levels are higher than those with heart function grade II heart failure(P<0.05),the difference is statistically significant;(3)The serum Gal-3 and inflammatory response factor hs-CRP in the heart failure group TNF-α and IL-6 levels are positively correlated with LVESD and LVEDD(r=0.473～0.558,P<0.01),and negatively correlated with LVEF(r =-0.825～-0.739,P<0.01);(4)MACE patients Serum Gal-3 and inflammatory response factors hs-CRP,TNF-α and IL-6 levels were higher than those of patients without MACE(P<0.05);(5)The receiver operating curve showed that the area under the ROC curve for Gal-3 diagnosis of HF was 0.864(P<0.05).When the cutoff value is83.19,the sensitivity is 78% and the specificity is 84.3%.Clinical randomized controlled study:(1)Compared with the control group,the Gal-3and NLRP3 levels of the treatment group were lower than those of the control group.Both groups can reduce the levels of PCP and PIIINP.Compared with the control group,the treatment group has a better effect;(2)The treatment group and the control group can reduce the indicators of heart LVEDD,LVESD,LVM,LVMI,LVEDVI and LVESVI,and increase the LVEF level.Among them,the treatment group was better than the control group in improving LVEF,LVESD,LVM,LVMI,LVEDVI and LVESVI(P<0.05);(3)Both groups could increase the 6-minute walking distance and reduce the Minnesota quality of life score,but the treatment group was excellent In the control group(P<0.05);(4)Both groups can reduce the levels of inflammatory factors TNF-α,IL-6,hs-CRP;in improving TCM syndromes,the treatment group is better than the control group,especially in improving wheezing The effects of shortness of breath,fatigue and cold limbs were more significant(P<0.05);Animal experiments:(1)Two rats in the model group and LQF-H group died,one in the low-dose Luqi Fang group and one in the MCP group.The appetite of the rats in the model group was significantly reduced after operation,the hair lacked gloss,the activity was reduced,the body weight was reduced,the urine output was significantly reduced,and there were varying degrees of edema.The above-mentioned situation of the rats in each medication group was improved compared with the previous one.(2)Compared with the sham operation group,cardiomyocyte necrosis,inflammatory cell infiltration,and collagen fiber proliferation were seen in the model group in cardiac staining;compared with the model group,cardiomyocyte necrosis,inflammatory cell infiltration and collagen fiber proliferation in the MCP and LQF groups All have been reduced to a certain extent.(3)After 4 weeks of drug intervention,compared with the sham operation group,the EF value of the model group decreased significantly;compared with the model group,the EF value of the LQF-L,LQF-M,LQF-H and MCP groups increased significantly.Compared with the MCP group,LVEF in the LQF-M and LQF-H groups increased significantly.Compared with the sham operation group,the FS value of the model group was significantly lower.Compared with the model group,the FS value of the LQF-L,LQF-M,LQF-H and MCP groups increased significantly(P<0.01).Compared with the MCP group,the FS of the LQF-M and LQF-H groups increased significantly.Compared with LQF-L,the FS of LQF-M and LQF-H groups increased significantly,but there was no statistical difference between the two.(4)Compared with the sham operation group,the Gal-3 protein expression in the model group was significantly increased(P<0.001).Compared with the model group,after drug intervention,Gal-3 in the LQF-L,LQF-M,LQF-H and MCP groups-3 protein expression decreased(P<0.01).Compared with the LQF-L group,the Gal-3 protein expression in the LQF-M,LQF-H and MCP groups decreased(P<0.01).Compared with the MCP group,the Gal-3 protein expression of the LQF-H group decreased(P<0.01),and the Gal-3 protein expression of the LQF-M(P=0.999)group was not different from that of the MCP group(P>0.05).Compared with the sham operation group,the protein expression of NLRP3 in the model group was significantly increased,which was statistically significant(P<0.01);compared with the model group,the protein expression of NLRP3 in the LQF-L,LQF-M,and LQF-H groups decreased After the intervention of MCP,the decrease of NLRP3 was more significant,with statistical difference(P<0.01).Compared with the control group,the expression of Caspase-1protein in the model group was significantly increased(P<0.001);compared with the model group,after adding Gal-3 inhibitor MCP and Luqi Fang,the expression level of Caspase-1 was significantly decreased.It was statistically significant(P<0.001),but the LQF-M and LQF-H groups were not statistically significant compared with the MCP group(P>0.05).Compared with the control group,the IL-1β protein expression in the model group increased significantly(P<0.001),indicating that the inflammation in the model group was further intensified;compared with the model group,after the intervention of MCP and Luqi Fang,LQF-M,LQF-H and MCP group IL-1β protein expression decreased(P<0.01),LQF-L(P=0.3770)was not statistically significant,and compared with MCP group,LQF-H group IL-1β decreased more Obviously(P=0.0112).(5)Compared with the sham operation group,the expression of Gal-3m RNA in the model group was significantly increased(P<0.001).Compared with the model group,the expression of Gal-3m RNA in the LQF-M,LQF-H and MCP groups decreased after drug intervention(P<0.001).Compared with the LQF-L group,the Gal-3m RNA expression in the LQF-H(P =0.0059)and MCP groups decreased(P<0.001),and there was no statistical significance in the LQF-M group.The expression of Gal-3m RNA in the LQF-M and LQF-H groups was not different from that of the MCP group(P>0.05).Compared with the sham operation group,the expression level of NLRP3 m RNA in the model group(P<0.001)was significantly higher.Compared with the model group,the NLRP3 m RNA content in the LQF-L,LQF-M,LQF-H and MCP groups was somewhat higher.Compared with the MCP group,the NLRP3 m RNA content of the LQF-H group decreased(P=0.0192<0.05),but the LQF-M and MCP groups were not statistically significant(P=0.0715).Compared with the sham operation group,the m RNA content of Ca Pase-1 in the model group increased significantly(P<0.001);compared with the model group,the Capase-1 m RNA content of the LQF-L,LQF-M,LQF-H and MCP groups Compared with the LQF-L group,the Ca Pase-1m RNA content of the LQF-H group decreased(P=0.0006<0.001),but the LQF-L,LQF-M and LQF-H groups were not statistically significant compared with the MCP group(P>0.05).Compared with the sham operation group,the IL-1β m RNA content in the model group was significantly increased(P<0.001);compared with the model group,the IL-1β m RNA content in the LQF-L,LQF-M,LQF-H and MCP groups was somewhat Decrease;compared with the LQF-L group,the levels of IL-1βm RNA in the LQF-M,LQF-H and MCP groups decreased(P<0.001),but there was no statistical significance between the LQF-H group and the MCP group(P>0.05)(6)The results of immunohistochemistry showed that compared with the sham operation group,the myocardial tissue of the model group showed changes in ventricular wall thickening,connective tissue hyperplasia,and increased Gal-3 expression.After the treatment of Luqi Decoction,the expression of Gal-3 decreased compared with the model group,indicating that the intervention of Luqi Decoction can inhibit the expression of Gal-3,which is mutually corroborated by the results of clinical studies.In the sham operation group,a small amount of brown-yellow type Ⅰ and Ⅲ collagen fibers were expressed.In the model group,a large number of irregularly arranged,brown-yellow type I and III collagen fibers can be seen.Compared with the model group,the collagen fibers in the LQF group and the MCP group were reduced.Compared with MCP,the LQF-H group is more effective in reducing the number of myocardial type I and III collagen fibers and improving the disorder of arrangement.(7)Compared with the sham operation group,the serum content of TNF-α in the model group was significantly increased(P<0.001);compared with the model group,the difference in LQF-L(P=0.0613)was not statistically significant.The LQF-M,LQF-H group and The content of TNF-α in the MCP group decreased(P<0.001);compared with the MCP group,there was no difference in the content of TNF-α protein in the LQF-L,LQF-M and LQF-H groups,but the LQF-H and LQF-L groups were Statistically significant(P=0.0379).Compared with the sham operation group,the serum level of IL-18 in the model group was significantly increased;compared with the model group,the serum level of IL-18 in the LQF-L,LQF-M,LQF-H and MCP groups decreased(P<0.001);and LQF-L group compared,LQF-H group IL-18 serum content decreased(P<0.001);compared with MCP group,LQF-L group was statistically significant(P=0.0005),LQF-M,LQF-H group had no statistics Learn meaning.Compared with LQF-L and LQF-M,the LQF-H group was statistically significant(P<0.001).The results of serum BNP showed that compared with the sham operation group,the BNP of the model group was significantly increased(P<0.001);compared with the model group,after the intervention of LQF and MCP,the BNP content decreased(P<0.05),but the LQF-H group was compared with MCP The difference between the groups was not statistically significant,and the effect was equivalent(P>0.05).Compared with the sham operation group,the serum level of IL-1βin the model group was significantly increased;compared with the model group,the serum level of IL-1β in the LQF-M,LQF-H and MCP groups decreased(P<0.001),LQF-L(P=0.3770)group;compared with LQF-L group,serum levels of IL-1β in LQF-H and MCP groups decreased(P<0.001),but there was no difference in LQF-M(P=0.7024);compared with MCP group,LQF-L,LQF-M,LQF-H group has statistical significance(P=0.0005);compared with LQF-L,LQF-M,LQF-H group has statistical significance(P<0.001).Conclusions: 1.Gal-3 and inflammation are positively correlated with chronic heart failure heart function classification,and negatively correlated with left ventricular ejection fraction.It is suggested that Gal-3 can diagnose and assist in the early diagnosis of chronic heart failure,which has important clinical significance.2.Luqi Decoction can reduce the levels of Gal-3,NLRP3,PICP,PIIINP in patients with chronic heart failure with Yang deficiency and blood stasis syndrome,reduce the levels of inflammatory factors TNF-α,IL-6,hs-CRP,and improve the symptoms and signs of patients in traditional Chinese medicine.Cardiac function classification and curative effect,improve the quality of life,delay myocardial fibrosis.3.The mechanism of Luqi Fang’s inhibition of myocardial fibrosis in rats with heart failure after myocardial infarction may be related to down-regulation of Gal-3 and NLRP3/IL-1β signaling pathways and inhibition of the release of pro-inflammatory factors TNF-α,IL-18 and IL-1β It is related to the down-regulation of the expression of PICP and PⅢNP. |
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