The Effect Of Tongmai Zhuyu Granule On The Methylation Of FoxO4,COL4A1_COL4A2 And SPHK1 Genes In Patients With Qi Stagnation And Blood Stasis Type Coronary Heart Disease

Objective: Based on the regulation of DNA methylation in the pathogenesis of coronary artery atherosclerotic heart disease(CHD)and Tongmai Zhuyu Granules significant clinical efficacy in the treatment of qi stagnant and blood stasis type CHD,it is hypothesized that Tongmai Zhuyu Granules can regulate the methylation modification of FoxO4,SPHK1 and COL4A1_COL4A2 genes in patients with qi stagnant and blood stasis type CHD and thus improve the clinical symptoms of CHD patients.To investigate the deeprooted regulatory factors,namely DNA methylation,in the treatment of qi stagnant and blood stasis type CHD with Tongmai Zhuyu granules.Methods: A total of 24 patients with qi stagnation and blood stasis type CHD attending the Heart Center of Qingdao TCM Hospital from October 2020 to October 2021 were selected by using randomized controlled trial.Randomly divided into control and test groups(12 in each group),and baseline data were recorded for each group of subjects.According to the Guidelines for the Primary Care of Non-ST-segment Elevation Acute Coronary Syndromes(2019),the control group was treated with conventional western medicine,and the test group was treated with Tongmai Zhuyu Granules on top of this treatment for a total of 28 days.To record and compare the scores of angina symptoms and Qi stagnation and Blood stasis symptoms before and after treatment in both groups to assess the efficacy of angina and Qi stagnation and Blood stasis symptoms.Methylation analysis of the promoter regions of FoxO4,SPHK1 and COL4A1_COL4A2 genes was performed before and after treatment in both groups of subjects based on sodium bisulfite sequencing(BSP).Results: 1 Clinical efficacyThe control group and the test group were comparable at baseline.1.1Points of angina symptoms and Chinese medicine symptoms: Both groups had significantly lower scores than before treatment(P < 0.01);and the test group was lower than the control group(P < 0.01)1.2 Angina pectoris efficacy: 66.7% in the control group compared to 91.7% in the test group(P < 0.05).1.3 Chinese medical evidence efficacy: The total effective rate of the control group was 25%,while the effective rate of the test group was 83.3%(P<0.01)2 FoxO4,SPHK1 and COL4A1_COL4A2 genes methylation analysisThe control group was comparable to the test group at baseline.There was no significant difference in the degree of promoter methylation of FoxO4,SPHK1 and COL4A1_COL4A2 genes before and after treatment in the control group(P>0.05).The level of FoxO4 gene promoter methylation in the test group after treatment was low compared with that before treatment(P < 0.01);the level of SPHK1 and COL4A1_COL4A2 gene promoter methylation in the test group after treatment was high compared with that before treatment(P < 0.001).Conclusion: 1.Tongmai Zhuyu Granules can improve the clinical symptoms of CHD patients and improve the symptoms of Qi stagnation and blood stasis2.The methylation level of FoxO4,SPHK1 and COL4A1_COL4A2 genes could be regulated by Tongmai Zhuyu Granules,which may be the treatment mechanism of Tongmai Zhuyu Granule in the patients with qi stagnation and blood stasis type CHD.