| Objective:Using flow cytometry (Flow cytometry, FCM) and enzyme-linked immunosorbent assay (ELISA), We accurately determine the regulatory T cells in peripheral blood (CD4+CD25+Foxp3+Treg cells),cytokine interleukin-10(IL-10) and transforming growth factor-β1(TGF-β1) levels in patients with heart failure of coronary artery disease, furthermore we study these factors with Shenfu injection to see their changes in order to provide experimental basis for clinical application in promoting qi and yang medicine,explore the mechanism of immune and inflammatory chronic congestive heart failure, and provide new ideas for the clinical treatment of chronic congestive heart failure.Methods:Patients with CHD were enrolled from the Department of Cardiovascular of Guang’an Men hospital from August2013to February2014.93enrolled patients with CHF were distributed in the study group and were evaluated by doctor in charge according to the NYHA functional classes well as the TCM syndrome differentiation.32enrolled CHD patients without CHF were in the control group. According to the standard of heart-kidney yang deficiency of heart failure,the patients in NYHA Ⅲ and IV were distributed to heart-kidney yang deficiency group(XSYX group), non-heart-kidney yang deficiency group (FXSYX group).The patients in heart-kidney yang deficiency group were randomly assigned to treatment with Shenfu injection or conventional western medicine treatment,the treatment continues about10±2days. All the serum samples of the enrolled patients were tested by Enzyme-Linked Immunosorbent Assay(ELISA) to determine the level of IL-10and TGF-β1, Chemiluminescence Immunoassay(CLIA) to test the level of NT-proBNP,and flow cytometry to test the ratio of regulatory T cells(CD4+CD25+Foxp3+Treg cells).The collected data was processed and analyzed by the statistical analysis software. The collected data was processed and analyzedby the statistical analysis software.Results:The study included93patients with heart failure (NYHA Ⅱ25cases,NYHA Ⅲ35cases,NYHAⅣ33cases)and32patients with non-heart failure (NYHA I). According to TCM syndrome differentiation, We divided NYHAIII-IV patients into two groups-56cases of heart-kidney yang deficiency and30cases of heart-kidney yang deficiency.After doing statistical analysis,there are no statistically significant differences between heart failure and non-heart failure groups at the level of age,gender and other aspects. The two groups XSYX and FXSYX are the same with the heart failure and non-heart failure groups.1.The serum levels of NT-proBNP and IL-10of heart failure patients are significantly higher than the non-heart failure patients,while the CD4+CD25+Foxp3+Treg cells ratio and the serum level of TGF-β1are significantly decreased compared with the non-heart failure patients (P<0.05);2.The serum level of NT-proBNP of heart failure patients with heart-kidney yang deficiency are significantly higher than the non-heart-kidney yang deficiency,while their the proportion of CD4+are significantly lower than the non-heart-kidney yang deficiency (P<0.05).3. The efficacy of Shenfu treatment to the symptom of patients with heart failure:Control group6cases (20.00%),effective in15cases (50.00%),8cases (26.67%),one case of aggravated (3.33%).Treatment group7cases (26.92%),effective in17cases (65.38%),2cases (7.69%), increased0cases (0%). The total efficiency of Control group are70.00%compared to the treatment group of92.31%,which there are statistically significant differences between the two groups.(P<0.05)4. The efficacy of Shenfu treatment to heart-kidney yang deficiency of patients with heart failure:Control group that were4cases (13.33%), effective in15cases (50.00%),9cases (30.00%), adding two cases (6.67%). Treatment group9cases (34.62%), effective in16cases (61.54%),1patient (3.85%), increased0cases (0%). The total efficiency of Control group are63.33%compared to the treatment group of96.15%,which there are statistically significant differences between the two groups.(P<0.05)5. The impact of Shenfu injection to the serum level of NT-proBNP,IL-10,TGF-β1and the ratio of CD4+CD25+Foxp3+Treg cell of patients with heart-kidney yang deficiency syndrome of chronic heart failure:Compared with initial treatment, the serum level of NT-proBNP and IL-10of both control group and treatment group decrease after Shenfu treatment,and there is a significant difference(P<0.05), meanwhile the ratio of peripheral blood CD4+CD25+Foxp3+Treg cells and the serum of TGF-β1increase,and there is a significant difference statistically difference (P<0.05). After treatment,the treatment group of the degree of reduced serum level of NT-proBNP and IL-10is more obvious than the control group, with a statistically significant differences (P<0.05), and the increased level of CD4+CD25+Foxp3+Treg cells ratio and TGF-β1also significantly higher than the control group, with a statistically significant differences.(P<0.05).Conclusion:1. There is a decreased level of CD4+CD25+Foxp3+Treg cells in patients with chronic heart failure,indicating the presence of immune activation status in patients with chronic heart failure.2. The serum level of TGF-β1was lower than the control group,and showed the tendency of decreasing with the rise of the NYHA functional class.3. The serum level of IL-10was higher than the control group,and showed the tendency of elevating with the rise of the NYHA functional class.4. The ratios of CD4+cells and CD4+CD25+Foxp3+Treg of the patients with heart-kidney yang deficiency syndrome of chronic heart failure are lower than non-heart-kidney yang deficiency syndrome,indicating there is a more stronger status immune activation than non-heart-kidney yang deficiency syndrome.5. Shenfu injection has the impact of improving the heart function and the heart-kidney yang deficiency syndrome, indicating that the drug is an effective treatment of coronary artery disease and chronic heart failure and heart-kidney yang deficiency syndrome of coronary heart failure.6. The impact of Shenfu injection to chronic congestive heart failure can increase the ratio of CD4+CD25+Foxp3+Treg cells,the serum levels of TGF-β1,and reduce the serum levels of NT-proBNP and IL-10,in order to reduce immune inflammation, thereby improving the heart function and the syndromes of heart-kidney yang deficiency to achieve therapeutic effect. |
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