Clinical Efficacy And Mechanism Of Xiaozheng Poxue Therapy In The Treatment Of Diabetic Lower Extremity Arterial Disease

Chapter Ⅰ Based on the real world study to explore the influencing factors of the prognosis of diabetic lower extremity arterial disease and the clinical efficacy of Xiaozheng Poxue therapyObjectiveUsing real-world research methods,the clinical efficacy and risk factors affecting the prognosis of diabetic lower extremity arterial disease treated with traditional Chinese medicine(TCM)for Xiaozheng Poxue therapy in the real world were evaluated.MethodsA retrospective cohort study was conducted to study diabetic lower extremity arterial disease.The patients were divided into two groups according to whether they accepted Xiaozheng Poxue therapy:Xiaozheng Poxue group(XZ group)and non Xiaozheng Poxue group(NXZ group).The endpoint of lower extremity vascular disease was used as the outcome index,and the PS matching method was used to control the confounding factors.The Cox regression model,Kaplan-Meire survival curve and Log-rank test were used to evaluate the clinical efficacy of TCM in the treatment of diabetic lower extremity arterial disease and the risk factors affecting the prognosis of the disease.ResultsA total of 544 cases were included in the study,excluding 40 cases with incomplete clinical electronic medical records and follow-up data.A total of 504 cases were analyzed.The average follow-up time was 45.52 months,and 160 patients had endpoint events.Before propensity matching,Cox regression model analysis showed that gender,age,smoking,history of hypertension,history of cardiovascular and cerebrovascular diseases,lower extremity symptoms,low density lipoprotein,glycosylated hemoglobin,degree of lower extremity vascular stenosis,statin therapy,antithrombotic therapy and other variables had no significant correlation with the composite endpoint of lead(P>0.05).The duration of diabetes,diabetic peripheral neuropathy,and albuminuria grade were closely related to the composite endpoint events(P<0.05).Gender,age,smoking,duration of diabetes,history of hypertension,history of cardiovascular and cerebrovascular diseases,diabetic peripheral neuropathy,lower extremity symptoms,albuminuria A2,degree of lower extremity vascular stenosis,statin therapy,antithrombotic therapy and other variables had no significant correlation with the cardiovascular and cerebrovascular end points of lead(P>0.05).Low density lipoprotein,glycosylated hemoglobin and albuminuria grade A3 were closely related to the occurrence of cardiovascular and cerebrovascular endpoint events(P<0.05).Before propensity score matching,Cox regression model analysis showed that the use of Xiaozheng Poxue Fang was 0.67 times higher than that of patients without Xiaozheng Poxue Fang(95%CI:0.47-0.95,P=0.025).In terms of cardiovascular and cerebrovascular end-point events,there was no significant correlation between Chinese medicine for eliminating symptoms and breaking blood and end-point events(P>0.05).Kaplan Meier survival curve showed that the traditional Chinese medicine of Xiaozheng Poxue therapy could delay the occurrence of endpoint events.After propensity score matching,variables such as gender,age,smoking,history of hypertension,history of cardiovascular and cerebrovascular diseases,diabetic peripheral neuropathy,lower extremity symptoms,low density lipoprotein,glycosylated hemoglobin,A3 grade of albuminuria,degree of lower extremity vascular stenosis,statin therapy and antithrombotic therapy had no significant correlation with the composite endpoint of lead(P>0.05).The duration of diabetes and the A2 grade of albuminuria were closely related to the occurrence of composite endpoint events(P<0.05).Gender,age,smoking,duration of diabetes,history of hypertension,lower extremity symptoms,low-density lipoprotein,glycosylated hemoglobin,albuminuria A2,lower extremity vascular stenosis,statin therapy and other variables were not significantly correlated with the cardiovascular and cerebrovascular end-point events of lead(P>0.05).Cardiovascular and cerebrovascular disease history,diabetic peripheral neuropathy,albuminuria A3 grade,antithrombotic therapy were closely related to cardiovascular and cerebrovascular end-point events(P<0.05).After the propensity score was matched,Cox regression model analysis showed that the patients who used Xiaozheng Poxue therapy were 0.62 times higher than those who did not use Xiaozheng Poxue therapy(95%CI:0.40-0.96,P=0.032).In terms of cardiovascular and cerebrovascular end-point events,there was no significant correlation between Chinese medicine for eliminating symptoms and breaking blood and end-point events(P>0.05).Kaplan Meier survival curve analysis showed that the traditional Chinese medicine of Xiaozheng Poxue therapy could delay the occurrence of endpoint events.ConclusionThe course of diabetes and albuminuria are risk factors for the progression of DLEAD.Antithrombotic therapy can reduce the risk of cardiovascular and cerebrovascular events in DLEAD.Traditional Chinese medicine can improve the prognosis of diabetic lead and delay the occurrence of diabetic lead endpoint events.Chapter Ⅱ Identification of Chemical Components of Xiaozheng Poxue Fang Based on Q-Orbitrap High Resolution Liquid Mass SpectrometryObjectiveThe study plans to analyze and identify the chemical components in Xiaozheng Poxue Fang by mass spectrometry using Q-Orbitrap high resolution liquid chromatography-mass spectrometry,so as to provide material basis for exploring the mechanism of Xiaozheng Poxue Fang in treating diabetes LEAD.MethodsQ-Orbitrap High Resolution Liquid Mass Spectrometry MethodResultsA total of 29 main active ingredients were identified,including DL arginine,d-(+)proline and other amino acid compounds;Nicotinic acid and other vitamin compounds;Tanshinone and other phenanthraquinones;Coumarin compounds such as subcoumarin and nucleotide compounds such as 5’-adenosine monophosphate and hypoxanthine..ConclusionThrough the identification and analysis of the main effective components of Xiaozheng Poxue Fang,it is suggested that the prescription may play its role in anti atherosclerosis by regulating lipid metabolism,inhibiting inflammatory reaction,improving vascular endothelial function,anti thrombosis,anticoagulation and other mechanisms.Chapter Ⅲ Exploring the mechanism of Xiao zheng Poxue Fang in the treatment of lower limb angiopathy in diabetes based on network pharmacologyObjectiveThis study is based on network pharmacology to explore the key targets of Xiaozheng Poxue Fang for the treatment of lower limb vascular disease in diabetes,and to provide basis for the follow-up study on the mechanism of Xiaozheng Poxue Fang for the treatment of lower limb vascular disease in diabetes.MethodsAll chemical components of Xiaozheng Poxue Fang were retrieved by tcmsp,CNKI,VIP and other platforms.The active ingredients of Xiaozheng Poxue Fang were screened according to oral bioavailability and drug properties.Via genecards ? Database and the integrated database of human genes and gene phenotypes(OMIM)were used to screen the gene and protein targets related to diabetes lead and establish the disease target data set.The obtained drug component targets and disease targets were mapped to each other,and the Venn diagram was made.Then,Cytoscape 3.7.2 software was used to construct the "drug component action target" network.The string database was used to construct protein-protein interaction network(PPI).David database was used for go biological process analysis and KEGG pathway analysis,and the network diagram of core active ingredient core target metabolic pathway was constructed.Autodock Vina(1.1.2)was used for molecular docking of active ingredients and key targets to verify their interaction activity.ResultsThe study found that the core targets of Xiaozheng Poxue Fang in the treatment of diabetic lower extremity arterial disease were IL6,STAT3,TNF,TP53,AKT1,GAPDH,EGFR,PTGS2,Jun,MMP9,etc.Go analysis showed that there were 32 items related to cell composition(CC)and 54 items related to molecular function(MF).The biological functions such as negative regulation of cell apoptosis,inflammatory response,hypoxia response,negative regulation of gene expression,and cell apoptosis process may be related to the treatment of diabetic lead with Xiaozheng Poxue Fang.KEGG pathway enrichment analysis showed that lipid and atherosclerosis,IL-17 signaling pathway,TNF signaling pathway were related to the pathway of Xiaozheng Poxue Fang in the treatment of diabetic lower extremity arterial disease.Molecular docking technology showed that tanshinone IIA could bind to IL6,AKT1,TNF and other proteins;Luteolin binds to STAT3 and TP53 proteins.ConclusionThe possible mechanism of Xiaozheng Poxue Fang in treating diabetes lower limb vascular disease is to regulate inflammatory factors,including IL-6,TNF,etc.Its possible signal pathways are lipid and atherosclerosis,IL-17 signal pathway,TNF signal pathway,etc.Chapter Ⅳ mechanism of Xiaozheng Poxue Fang on diabetic rats with lower extremity arterial diseaseObjectiveIn this study,we observed the effect of Xiaozheng Poxue Fang on TNF in a rat model of diabetic lower extremity vasculopathy-α/NF κ Objective to explore the mechanism of Xiaozheng Poxue Fang in the treatment of diabetic LEAD.MethodsThe animal model of diabetic lower extremity arterial disease was established in SD rats.Twelve rats without modeling were used as the blank group.The successful rat model was randomly divided into model group,high-dose group,medium dose group,low-dose group and Western medicine group,with 12 rats in each group.The western medicine group was given aspirin,and the low-,medium-,and high-dose groups were given different doses of Xiaozheng Poxue Fang.The levels of CRP,ET,NO,oxLDL,TNF-α,IL-6 and IL-8 in rat serum were detected by ELISA.TNF-α,NF κ B,IKK β,INOS,BCL2,Bax,caspase-1 and caspase-3 mRNA were detected RT-PCR.Expression of TNF-α,IL-6,NF-κB,IKK β,IKK α,PTGS2 and iNOS protein were Detected by Western blot.ResultsELISA detection showed that:compared with the blank control group,the level of CRP,ET,NO,oxLDL,TNF-α、IL-6 and IL-8 were significantly increased(P<0.01);Compared with the model group,Xiaozheng Poxue Fang(high,medium and low dose groups)could reduce the levels of the above pro-inflammatory factors(P<0.01).RT-q PCR results showed that compared with the blank group,The mRNA expressions of TNF-α,NF κ B,IKK β,iNOS,Bax,caspase-1 and caspase-3 increased(P<0.05),while the expression of bcl2 mRNA decreased(P<0.05).Compared with the model group,The mRNA expression of TNF-α,NF κB,IKK β,iNOS,Bax,caspase-1 and caspase-3 in aspirin group and Xiaozheng Poxue Fang(low,medium and high dose groups)decreased(P<0.05),while the mRNA expression of BCL2 increased(P<0.05).Western blot results showed that compared with the blank group,The protein contents of TNF-α,IL-6,NFκB,IKK β,IKK a,PTGS2,iNOS in the model group increased(P<0.05);Compared with the model group,The protein contents of TNF-α,IL-6,NFκB.IKK β,IKK a,PTGS2 and iNOS in aspirin group and Xiaozheng Poxue Fang(low,medium and high dose groups)were decreased(P<0.05),and TNF-α,IL-6,NFκB,IKK β,IKK a,PTGS2 and iNOS in high dose group of Xiaozheng Poxue Fang were lower than those of Xiaozheng Poxue Fang low-dose group,mid-dose group and aspirin group(P<0.05).ConclusionXiaozheng Poxue Fang can inhibit CRP and TNF-α、IL-6,IL-8 and other inflammatory factors secrete,regulate ET,NO and other endothelial factors,up regulate BCL2 mRNA,down regulate Bax,caspase-1 and caspase-3 mRNA expression,so as to improve the inflammatory state of diabetic lead and protect vascular endothelial function.Its mechanism may be related to TNF-α/IKK β/NF-κB signaling pathway.