Study On The Correlation Between The Adventitia Vasa Vasorum Neovascularization And Macrophages As Well As Lymphocytes Recruitment In Type 2 Diabetic Patients With Lower Extremity Macroangiopathy And TCM Syndromes

Objectives:In this study,we focused on the femoral artery of patients with major amputation of type2 diabetic foot ulcers to observe the correlation between adventitia vasa vasorum neovascularization,macrophages,lymphocytes as well as the degree of macrovascular lesions,and to further explore the correlation between the relevant clinical indicators and TCM syndrome types.From the clinical and pathological point of view,we will explore prevention and treatment strategies to provide a new approach for diabetic macroangiopathy.Methods:A total of 55 patients with type 2 diabetes mellitus complicated with macrovascular disease and 6 autopsy cases without diabetes mellitus were enrolled in the study.They all underwent the middle and lower 1/3 amputation and were split into macrovascular disease group and control group.The baseline data such as general condition,diabetic complications and clinical indicators of the patients were compared between the two groups.Vascular pathological structures were observed by HE staining.Victoria blue staining was used to measure and analyze the arterial diameter,vascular wall thickness,intima-media thickness,adventitia thickness and the percentage of lumen stenosis(area stenosis).Immunohistochemistry(CD34,CD68,CD20,CD3)was used to detect the density of adventitia vasa vasorum neovascularization and the positive expression of macrophages,B lymphocytes as well as T lymphocytes.To explore the correlation between adventitia vasa vasorum neovascularization,macrophages,lymphocytes,as well as the correlation between different TCM syndrome types.Results:1.TC and LDL-c in diabetic macroangiopathy group were higher than controls,while HDL-c was lower than controls.(P < 0.05).There were no significant changes in gender,age,TG,CVD,BMI,hypertension history,smoking history,cerebrovascular disease,statin use history and vascular intervention history between the two groups(P > 0.05).2.In the diabetic macroangiopathy group,the pathological and structural changes mainly consisted of moderate or severe stenosis of the arterial lumen,degeneration and necrosis of local tissue damage in the intimal layer of the vessel,thrombus tissue appearing in the lumen,muscularization and structural disintegration of the thrombus tissue,as well as degeneration and necrosis of the tissue within the thrombus.In the middle membrane layer,the local middle membrane smooth muscle necrosis,inflammatory cell infiltration(mainly neutrophils and macrophages),vacuolar degeneration and mineraloid degeneration in the necrotic area(dark blue).Part of the adventitia degenerated and necrotized with the middle membrane,accompanied by structural incompleteness,interstitial inflammatory cell infiltration and dense adventitia vasa vasorum neovascularization.In the control group,there were no obvious pathological changes in the intima and adventitia,mainly the pathological structural changes of local middle membrane tissue degeneration and necrosis.3.The vascular wall thickness,intima-media thickness and the percentage of lumen stenosis(area stenosis)measured by morphology in the diabetic macroangiopathy group were higher than controls(P < 0.05).There was no significant difference in arterial diameter and adventitia thickness(P > 0.05).4.Compared with the control group,the density of adventitia vasa vasorum neovascularization and the positive expression of CD3,CD20,and CD68 were significantly higher in the diabetic macroangiopathy group(P < 0.05).5.In different degree of diabetic macroangiopathy group,with the aggravation of AS severity,the density of adventitia vasa vasorum neovascularization as well as the positive expression of CD20 and CD68 increased significantly(P < 0.05).There was no significant difference in the positive expression of CD3(P > 0.05).6.In the density of adventitia vasa vasorum neovascularization quartile subgroups of diabetic macroangiopathy group,the overall positive expression of CD3,CD20,and CD68 was gradually increased as the density of adventitia vasa vasorum neovascularization increased(P< 0.05).7.There was a positive correlation between the density of adventitia vasa vasorum neovascularization and the average positive expression of CD68(r=0.62)and CD20(r=0.59)in diabetic macroangiopathy group(P < 0.05).There was no correlation with the average positive expression of CD3(r=0.19,P > 0.05).8.Compared with the group of Dampness and Heat in Spleen and stagnant blood,the thickness of vascular wall,intima-media thickness and lumen area were significantly increased in the group of Qi Yin asthenia and stagnant blood as well as the group of Ying Yang asthenia and stagnant blood(P < 0.05).Compared with the group of Yin asthenia endogenous heat and stagnant blood,the thickness of vascular wall,intima-media thickness and lumen area stenosis in the group of Qi Yin asthenia and stagnant blood and the group of Ying Yang asthenia and stagnant blood were significantly increased(P < 0.05).There was no significant difference in arterial diameter and adventitia thickness among the four syndrome types(P > 0.05).9.Compared with the group of Dampness and Heat in Spleen and stagnant blood,the density of adventitia vasa vasorum neovascularization and the positive expression of CD68 and CD20 were significantly increased in the group of Qi Yin asthenia and stagnant blood as well as the group of Ying Yang asthenia and stagnant blood(P < 0.05).Compared with the group of Yin asthenia endogenous heat and stagnant blood,the density of adventitia vasa vasorum neovascularization and the positive expression of CD68 and CD20 in the group of Qi Yin asthenia and stagnant blood as well as the group of Ying Yang asthenia and stagnant blood were significantly increased(P <0.05).The positive expression of CD3 had no significant change among the four syndrome types(P > 0.05).Conclusions:1.There is a large number of adventitia vasa vasorum neovascularization in the adventitia of type 2 diabetic macroangiopathy,and positively correlated with inflammatory reaction and AS lesions.2.Patients with macroangiopathy in type 2 diabetes with different TCM syndromes have different degrees of lesion.Patients with Qi Yin asthenia and stagnant blood as well as Ying Yang asthenia and stagnant blood syndrome had more severe AS lesions,higher density of adventitia vasa vasorum neovascularization,and higher positive expression of macrophages and B lymphocytes.