| This research subject is based on the cross-cutting topic "Evidence study on the treatment of diabetes by Shenqi Jiangtang Granules,a traditional Chinese medicine for supplementing Qi and spleen and tonifying kidney".It mainly includes two parts:clinical study of "Syndrome characteristics of spleen and kidney qi deficiency syndrome of type 2 diabetes complicated with coronary heart disease,clinical characteristics of blood circulation reconstruction population" and experimental study of "mechanism of action of Shenqi Jiangtang Granules to improve myocardial cell injury".The results of the first part of clinical study showed that the spleen and kidney qi deficiency were more severe in patients with diabetes complicated with coronary heart disease due to revasectomy,and supplementing Qi and spleen and tonifying kidney was the main treatment.Shenqi Jiangtang Granule was a drug with high frequency of use.Based on the results of the first part of clinical study,rat H9C2 cells were selected for experimental study.Objective:To explore the therapeutic mechanism of Shenqi Jiangtang Granules on diabetes mellitus complicated with coronary heart disease.1 Clinical StudyObjective:To investigate the clinical characteristics of syndrome in patients with type 2 diabetes complicated with spleen and kidney qi deficiency syndrome of coronary heart disease,and analyze the related factors affecting glycated hemoglobin,glycated albumin and TCM syndrome scores.To explore the clinical characteristics of type 2 diabetes mellitus complicated with coronary heart disease revascularization group,and to provide ideas for the clinical diagnosis and treatment of diabetes complicated with coronary heart disease.Methods:1.Based on the ward electronic medical record system,this study collected the relevant data of type 2 diabetes patients with spleen and kidney qi deficiency syndrome of coronary heart disease from September 2020 to March 2023 in the Endocrinology and cardiovascular departments of Guang ’anmen Hospital,including demographic data,basic medical history,laboratory examination and statistical analysis,to explore the demographic data characteristics of patients.2.TCM syndrome scores of patients were recorded,and univariate analysis and Logistic regression analysis were used to investigate the correlation between spleen and kidney qi deficiency syndrome in patients with type 2 diabetes complicated with coronary heart disease and islet function,blood glucose,glycated hemoglobin,glycated albumin,common blood lipids and common cardiovascular risk factors.Excel software was used to calculate the syndrome score of spleen and kidney qi deficiency,and the correlation between the syndrome score and related laboratory indexes and examinations was observed,so as to provide reference for the diagnosis and treatment of spleen and kidney qi deficiency syndrome of diabetes complicated with coronary heart disease.3.According to whether revascularization was performed or not,type 2 diabetes mellitus complicated with CHD was divided into revascularization group and non-revascularization group.Clinical data were collected to analyze their clinical characteristics.Results:1.Clinical features:(1)Gender distribution:Among the 100 patients,60 were male and 40 were female;(2)Age distribution,diabetes,coronary heart disease course characteristics:the number of 60-74 years old was the largest,56 cases(56%);The number of patients≤59 years old was followed by 30 cases(30%);The minimum number of patients aged≥75 years was 14(4%).There was statistical significance in the composition of type 2 diabetes mellitus complicated with CHD in men and women.(3)BMI mainly distributed in overweight and obesity.(4)Diabetic complications,diabetic peripheral vascular disease>Diabetic peripheral neuropathy>Diabetic abnormal sweating function>Diabetic retinopathy>Diabetic nephropathy>Diabetic Gastroparesis>Diabetic foot.The highest proportion of diabetic peripheral vascular disease was 73%.(4)Arteriosclerosis,the most common arteriosclerosis is carotid artery and lower limb artery,accounting for 60%,and the main manifestation is atherosclerosis with plaque.(5)The number of patients with hyperlipidemia was the largest with a prevalence of 92%.(6)In this study,coronary artery disease was diagnosed by coronary angiography,and percutaneous coronary intervention(PCI)and coronary artery bypass grafting(CABG)accounted for 47%and 3%of the total number ofpatients,respectively.2.Syndromic characteristics:the symptoms and signs with frequency greater than 80%were ranked from high to low,respectively,palpitation 97%,chest tightness 96%,eating less and staying 90%,mental fatigue 89%,waist and knee tenderness 83%,chest pain 86%,shortness of breath and lazy speech 84%,insomnia 83%.Tongue often see tongue dark red,light dark,fat fur with teeth marks,white and greasy fur;The pulse is usually string,smooth and thin.(1)The score of spleen and kidney qi deficiency syndrome in female was higher than that in male,with significant statistical difference;(2)TCM syndrome scores increased with age,but there were significant statistical differences among different age groups;(3)With the increase of the duration of diabetes,the syndrome score of patients showed an increasing trend,and the duration of diabetes>10 years was significantly different from the duration of diabetes<5 years and the duration of diabetes 5-10 years.(4)With the increase of duration of coronary heart disease,patients’ syndrome scores showed an increasing trend.The duration of coronary heart disease<5 years was significantly different from that of coronary heart disease 5-10 years and that of coronary heart disease>10 years.3.Correlation Analysis(1)Spearman correlation study:①Correlation of TCM syndrome scores:age,diabetes course,coronary heart disease course,total number of complications,FPG,2hPG,glycated hemoglobin,glycated albumin,ESR were significantly positively correlated with TCM syndrome scores;systolic blood pressure,whole blood troponin,MA/CR,HOMA-IR were positively correlated with TCM syndrome scores,and the difference was statistically significant.FC-P,2hPC-P,EF were negatively correlated with TCM syndrome scores,while total cholesterol,lipoprotein(a),serum homocysteine were negatively correlated with TCM syndrome scores,and the difference was statistically significant.② Correlation of glycated hemoglobin:diabetes course,coronary heart disease course,fasting blood glucose,2hPG,FINS,glycated albumin,HOMA-IR were significantly positively correlated with glycated hemoglobin,while the total number of complications,ESR were positively correlated with glycated hemoglobin and 24-hour urinary protein quantitation,and the difference was statistically significant.FC-P,2hPC-P,ApoA I showed significant negative correlation with glycosylated hemoglobin.③ Glycated albumin correlation:diabetes course,coronary heart disease course,total complications,FPG,2hPG,FINS,glycated hemoglobin,HOMA-IR,ESR were significantly positively correlated with glycated albumin,and lactate dehydrogenase was positively correlated with glycated albumin,the difference was statistically significant.FC-P,2hPC-P,TC,ApoA I,lp(a)were negatively correlated with glycated albumin,and EF was negatively correlated with glycated albumin,the difference was statistically significant.(2)Regression analysis:diabetes course,coronary heart disease course,fasting C-peptide,2-hour C-peptide,TCM syndrome score,erythrocyte sedimentation rate were the influencing factors of substandard HBA1C.TCM syndrome score and 2-hour C-peptide are important factors affecting the substandard glycated hemoglobin.TCM syndrome score(OR=0.559,95%CI confidence interval 0.427-0.733,P=0.000),2-hour C-peptide(OR=4.518,95%CI confidence interval 1.064-19.191,P=0.041).4.Clinical characteristics of T2DM combined with coronary artery disease spleen and kidney qi deficiency syndrome revascularization group:Type 2 diabetes combined with coronary artery disease spleen and kidney qi deficiency syndrome were divided into revascularization group and non-revascularization group according to whether revascularization was performed,and relevant data were analyzed.(1)Syndrome score:TCM syndrome score of revascularization group was higher than that of non-revascularization group,and the difference was statistically significant.(2)Blood glucose:GA,2hPG and FINS were higher in the revasectomy group than in the non-revasectomy group,and FC-P and 2hPC-P were lower,with statistical significance.(3)Myocardial enzyme and EF values:CK,LDH,LDH1 and CK-MB in revascularization group were higher than those in nonrevascularization group,and the difference was not statistically significant.MYO and BNP were higher in revascularization group than in non-revascularization group,and the difference was statistically significant.(4)Blood lipid:total cholesterol,LDL-C and ApoB in the revascularization group were lower than those in the non-revascularization group,and the difference was statistically significant.(5)Biochemistry:UA and HCY in the revascularization group were higher than those in the non-revascularization group,and the difference was statistically significant.(6)Urinary protein:MA/CR in revascularization group was higher than that in non-revas cularization group,and the difference was statistically significant.Conclusion:1.Clinical characteristics:The majority of type 2 diabetes patients with spleen and kidney qi deficiency syndrome of coronary heart disease are elderly,overweight and obese,and have a long course of disease.The complications are mostly diabetic peripheral vascular disease,carotid artery and lower limb artery are the common sites of arteriosclerosis,and hyperlipidemia is the most common complicated disease.2,Syndrome characteristics:Symptoms and signs with palpitations,chest tightness is common,tongue tongue more dark red,light dark,fat fur with teeth marks,white greasy fur,pulse more strings,smooth,fine.3.Analysis of related influencing factors:Diabetes course,coronary heart disease course,FC-P,2hPC-P,TCM syndrome score and erythrocyte sedimentation rate were the influencing factors for the substandard HBA1C,and TCM syndrome score and 2hPC-P were the main influencing factors.4.Characteristics of people with heavy vascular transport:(1)Syndrome score:The TCM syndrome score of the revascularization group was higher than that of the nonrevascularization group.(2)Blood glucose:GA,2hPG and FINS were higher in revasectomy group,while FC-P and 2hPC-P were lower.(3)Myocardial enzymes and EF values:MYO and BNP in revascularization group were higher than those in non-revascularization group.(4)Cholesterol:CHO in revascularization group was lower than that in non-revascularization group;The LDL-C and ApoB in revascularization group were lower than those in nonrevascularization group.(5)Biochemistry:UA and HCY in revascularization group were higher than those in non-revascularization group.(6)Urinary protein:MA/CR in revascularization group was higher than that in non-revascularization group.2 Experimental studyOBJECTIVE:To observe the effect of Shenqi Jiangtang Granules on improving H9C2 cell injury induced by high sugar combined with sodium palmitate,and to explore the possible mechanism of Shenqi Jiangtang Granules on improving myocardial cell injury from the perspective of inhibiting cell apoptosis.METHODS:(1)Establishment of H9C2 cell damage model:Cell damage was induced by 25mM high sugar combined with different concentrations of sodium palmitate(PA),and cell viability was calculated by CCK8 method to determine the modeling concentration.(2)Preparation of drug-containing serum and blank serum of Shenqi Jiangtang Granules:SD rats were given Shenqi Jiangtang granules(3.6g/kg)for 7 times by intragastric administration,blood was collected from abdominal aorta to prepare drug-containing serum.(3)Cytotoxicity test:CCK-8 method was used to evaluate the toxic effect of Shenqi Jiangtang Granule drug-containing serum on cells and determine the dosage of drug serum.(4)Experimental grouping and treatment:H9C2 cells were divided into blank control group,model group,Shenqi Jiangtang granule drug-containing serum high,medium and low dose groups.The corresponding drug intervention lasted for 24h,and the blank control group was treated with 10%blank serum.(5)Morphological observation:The morphological changes of H9C2 cells in each group were observed by inverted phase contrast microscope;(6)Apoptosis:Flow cytometry and immunofluorescence labeling were used to detect the number and percentage of apoptosis in each group,and the effect of Shenqi Jiangtang granules on apoptosis of cardiomyocytes was observed.(7)Oxidative stress:The effects of drug-containing serum of Shenqi Jiangtang granules on SOD and MDA were detected.(8)Myocardial protective factor gene expression levels:RT-PCR was used to detect the mRNA expression levels of LPA1 and LPA3,and the influence of drug-containing serum of Shenqi Jiangtang Granules on the mrna expression levels of LPA1 and LPA3 was observed.Results:(1)25mM high sugar combined with 450 μ M sodium palmitate induced H9C2 myocardial cell injury.(2)The appropriate concentration range of Shenqi Jiangtang Granule(SQ)medicated serum to interfere with H9C2 cell injury model was 2%-6%.(3)In the model group induced by high sugar combined with sodium palmitate,cardiomyocytes were blunt,their adhesion ability decreased,apoptotic cells were suspended from the wall,the growth rate was slow,and the cell space was enlarged.After intervention,the cell morphology of the low,middle and high SQ drug-containing serum group was improved,and the cell density was higher than that of the model group.(4)Flow cytometry showed that the apoptosis rate of myocardial cells in the model group was significantly increased compared with the normal group(P<0.05).Compared with the model group,SQ drug-containing serum group decreased the apoptosis rate of myocardial cells in the model group(P<0.05).(5)Hoest33342 staining showed that the nuclei of H9C2 cells in the normal control group were blue,uniformly stained,and the nuclei were round or oval.In the model group induced by high sugar and sodium palmitate,the nuclei showed signs of cell apoptosis,such as dense and dense staining,irregular contraction,fragmentation,etc.After the intervention of SQ concentration groups,the cell morphology was improved,and dense and dense and dense staining nuclei and cell fragments were reduced.(6)Compared with the normal group,SOD was decreased and MDA was increased in the model group;After the intervention of Shenqi Jiangtang granules,SOD increased and MDA decreased in H9C2 cells compared with model group.(7)Compared with the model group,the mRNA levels of LPA1 and LPA3 in H9C2 cells were significantly increased after the intervention of Shenqi Jiangtang Granules,with statistical significance(P<0.01).Conclusion:(1)The medicated serum of Shenqi Jiangtang granules can improve the nuclear morphology of cardiomyocytes,reduce cell debris and promote cell proliferation.(2)The drug-containing serum of Shenqi Jiangtang granules reduced the apoptosis rate and enhanced the antioxidant capacity of myocardial cells in the model group.(3)Shenqi Jiangtang Granules medicated serum can improve myocardial cell injury and inhibit cell apoptosis,which is related to the regulation of LPA1 and LPA3mRNA expression levels. |
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