| Objective:The prevalence rate of diabetes in China has increased rapidly from 9.7% in 2007 to 11.6% in 2010. The diabetes and its complications have seriously harmed the health of the people. The pathogenesis of type 2 diabetes is complex. The traditional concept thinks that it belongs to the category of metabolic diseases while modern research thinks that it is a chronic systemic inflammatory diseases. The polarization of monocyte/macrophage keeps up with chronic systemic inflammation, the main polarization forms are inflammatory M1 and anti-inflammatory M2. In 2010 Japanese Satoh et al found monocyte/macrophage of the obese type 2 diabetes mellitus had the presence of polarization imbalances, surface markers of inflammatory type M1 cells increased and surface markers of anti-inflammatory type M2 cells decreased. So this study speculates that polarization imbalance of monocyte/macrophage plays an important role in the pathogenesis of type 2 diabetes. In this study, we aimed to explore the relationship between monocyte/macrophage polarization imbalance with type 2 diabetes and its complications by analyzing the M1/M2 polarization state of peripheral blood monocytes with hospitalized type 2 diabetes patients in northern China. Method:1. This study used cross-sectional study. Type 2 diabetes mellitus patients(108 cases) in Metabolic Diseases Hospital of Tianjin Medical University and normal control volunteers(50 cases) in physical examination center of Tianjin Medical University Airport International Hospital were selected randomly. Then we collected general condition, biochemical indicators and imaging examination results of the subjects.2. In the early morning we got elbow venous blood of subjects and used the flow cytometry to detect the proportion of M1/M2 monocytes. The labeled antibodies of M1 monocytes contained CD68(leukocyte differentiation antigen 68) and CCR2(chemokine-C receptor-2) and M2 monocytes were labeled with CD163 and CX3CR1(chemokine-X3 C receptor-1). Then we calculated the ratio of M1/M2. Two independent sample t test was used to compare the proportion of type M1 and type M2 and the ratio of type M1/M2 in peripheral blood monocytes of patients with type 2 diabetes mellitus and control group. According to the complications of type 2 diabetes, all samples were divided into several subgroups, explored the relationship between monocyte/macrophage polarization imbalance and diabetic complications.3. Primary peripheral blood monocytes of type 2 diabetic patients and normal people were cultured in the condition of 37℃, 5% CO2. After 24 hours, cells state were stable,we began the intervention experiment. The experiment was divided into four groups: healthy control group(monocytes of healthy group), cases control group(monocytes of T2DM), healthy intervention group(monocytes of healthy group + 0.1mg/L LPS) and cases intervention group(monocytes of T2 DM + 0.1mg/L LPS), while cells were incubated in the condition of 37℃, 5% CO2 for 24 hours. We analyzed the changes of mononuclear cells polarization after inflammatory stimulation of with flow cytometry. Results:1. Compared with normal control group, patients with type 2 diabetes mellitus peripheral blood M1 monocytes percentage and M1/M2 ratio were higher than the normal control group while M2 monocytes percentage was lower than the normal control group. The difference was statistically significant(P<0.05).2. The proportion of peripheral blood M2 monocytes in macrovascular complications group and with both macrovascular and microvascular complications group was lower than that of the simple diabetes group(P<0.05). M1/M2 ratio with both macrovascular and microvascular complications group was higher than the simple diabetes group, macrovascular complications group and microvascular complications group(P<0.05).3. M1 monocytes proportion and M1/M2 ratio in patients with three diabetic macrovascular complications group were higher than the simple diabetes group, only one kind of macrovascular complications group and two kinds of macrovascular complications group, while M2 monocytes proportion in patients with three kinds of diabetic macrovascular complications was less than the simple diabetes group and only one kind of macrovascular complications group, the difference was statistically significant(P<0.05). M1 monocytes proportion and M1/M2 ratio in coronary heart disease group were higher than the simple diabetes group, while M2 monocytes proportion in coronary heart disease group was less than the simple diabetes group(P<0.05). M2 monocytes proportion in cerebrovascular disease group was less than the simple diabetes group, while M1/M2 ratio in cerebrovascular disease group was higher than the simple diabetes group(P<0.05). M2 monocytes proportion in lower extremity artery disease group was less than the simple diabetes group, while M1/M2 ratio in lower extremity artery disease group was higher than the simple diabetes group(P<0.05).4. M1 monocytes proportion and M1/M2 ratio in patients with three diabetic microvascular complications group were higher than the simple diabetes group, only one kind of microvascular complications group and two kinds of microvascular complications group, while M2 monocytes proportion in patients with three kinds of diabetic macrovascular complications was less than the simple diabetes group, only one kind of microvascular complications group and two kinds of microvascular complications group, but the difference has not yet been statistical significance(P > 0.05). M1 monocytes proportion in the early DN group and clinical DN group were higher than the DM group(P<0.05). M1 monocytes proportion and M1/M2 ratio in DR group were higher than the simple diabetes group, while M2 monocytes proportion in DR group was less than the simple diabetes group, but the difference was not statistically significant(P>0.05). M1 or M2 monocytes proportion and M1/M2 ratio between DPN group and the simple diabetes group had no differernce(P>0.05).5. M1 monocyte proportion and M1/M2 ratio in more than 10 years group of diabetic patients were higher than that of less than 10 years groupthe difference has statistical significance(P<0.05). But M2 monocytes proportion between two groups had no differernce(P>0.05).6. Through multiple linear regression analysis, the results showed that M1 monocytes proportion was related to diastolic blood pressure, D-dimer and hs-CRP, while M2 monocytes proportion and M1/M2 ratio were only related to the hs-CRP(P<0.05).7. With flow cytometry detectation after 0.1 mg/L LPS intervention for 24 hours, M1 monocytes proportion and M1/M2 ratio in healthy controls and diabetic patients increased and diabetic patients increased to a greater extent than healthy controls, while M2 monocytes proportion in healthy controls and diabetic patients decreased and diabetic patients decreased more than healthy controls, the difference was statistically significant(P<0.05). Conclusion:1.There was an imbalance of monocyte/macrophage M1/M2 polarization in patients with type 2 diabetes, while inflammatory type M1 cells increased and anti-inflammatory type M2 cells decreased. The polarization imbalance in patients with diabetic macrovascular complications and diabetic nephropathy was significantly, and patients with long course of the diseasethe were more obvious.2. Monocytes of patients with type 2 diabetes had the pre-inflammatory state, and inflammatory response was much stronger. Morever, monocyte/macrophage M1/M2 polarization imbalance associated with hs-CRP. There was a close contact among inflammation, monocyte/macrophage M1/M2 polarization imbalance and type 2 diabetes mellitus. |
PDF Full Text Request