| Background and objectiveThe prevalence of diabetic nephropathy(DN),one of important reasons which results in ESRD,is increasing year by year. The pathogenesis of DN is not fully understood as yet,more and more studies support that DN is a disease associated with immune inflammation. Th17cells and Treg cells are two important subsets of CD4+T cells,the imbalance of Th17and Treg cells is involved in many human autoimmune disorders,whether and how the imbalance of Th17and Tregs participates the pathogenesis of DN need to be further expounded.The curative effect of Modern Medicine to DN is not satisfied, the superiority of Traditional Chinese Medicine has been manifested but doctors’viewpoints are various,some researches of the relationship between syndrome of TCM of DN and laboratory index, special molecular biology index has been reported,but the correlation between syndrome of DN and the imbalance of Th17and Treg cells is not appeared.So we will observe the change of the balance of Th17and Tregs in the patients with DN;reveal the important immune inflammatory mechanisms of DN; find out the associativity of the change of the balance of Th17and Tregs along with the variation of syndrome types of DN and provide objective evidences to treat DN based on syndrome differentiation.Research methods15T2D adult patients,15adult patients diagnosed DN with microalbuminuria,15adult patients diagnosed DN with massive proteinuria and15healhy adults whose age and sex matched the patients were enrolled in this research.Urine and blood samples were collected to separately assay urine album-creatinine ratio and glycosylated hemoglobin,blood fat,serum creatinine and blood urea nitrogen;the frequence of Th17and Treg cells in peripheral blood were evaluated via flow cytometry,and the Th17/Treg ratio was calculated; plasma IL-17A,IL-10and TGF-betal were detected through ELISA.Then the correlation between Th17,Tregs, the Th17/Treg ratio,their correlated cytokines and urine album-creatinine ratio of patients with diabetic nephropathy was analyzed. Syndrome differentiation of DN patients was recorded,then the correlation between changes of the balance of Th17and Tregs,correlated cytokines and syndrome of TCM was analyzed.Results(1)The levels of HbAlC and blood fat are not significantly different between T2D and DN patients in our study,and they are uncorrelate with ACR.(2)The proportion of peripheral blood Th17,Th17related cytokines IL-17A level of adult patients with type2diabetes and diabetic nephropathy are significantly higher than these indexes of normal people(P<0.05),and positively correlate with ACR(r=0.531,P=0.003;r=0.527,P=0.003), DN patients with massive proteinuria is the highest,following the patients with microalbuminuria.(3)Compared with normal adults,Type2DN patients peripheral blood Treg ratio decreases significantly (P<0.05), and negatively correlates with ACR(r=-0.377,P=0.040). The peripheral blood Treg proportion of T2D patients and normal adults has no statistical difference (P>0.05), but also shows a downward trend, Tregs related plasma cytokines IL-10and TGF-beta1of DN patients are significantly increased compared to healthy ones(P<0.05),and they are positively correlated with ACR(r=0.420,P=0.021;r=0.613,P<0.001).The level of IL-10,TGF-betal in DN patients with massive proteinuria,the level of TGF-beta1in DN patients with microalbuminuria is higher than those of T2D patients(P<0.05),but there is no difference between DN with microalbuminuria and T2D patients’ level of IL-10(P>0.05).(4) The ratio of Th17/Treg cells in patients suffered from T2D,DN with microalbuminuria and DN with massive proteinuria manifests an increasing trend(P<0.05), and with urinary ACR positively correlated(r=0.663,P<0.001).(5)In DN patients with TCM principle syndrome, peripheral blood Th17cell ratio of patients with deficiency of Yang in the spleen and kidney is significantly higher than that of patients with deficiency of liver Yin and kidney Yin, deficiency of both qi and yin and deficiency of both Yin and yang (P<0.05), There is no statistical difference among patients with deficiency of liver Yin and kidney Yin, deficiency of both qi and yin and deficiency of both Yin and yang(P>0.05).(6)Patients with deficiency of Yang in the spleen and kidney have a significantly lower peripheral blood Treg cells percentage than patients with deficiency of both qi and yin and deficiency of both Yin and yang(P<0.05),while the Treg cells percentage of patients with deficiency of both Yin and yang is higher than that of patients with deficiency of liver Yin and kidney Yin and patients with deficiency of both qi and yin (P<0.05), there is no statistical difference between patients with other principle syndrome(P>0.05).(7)Compared with patients with deficiency of liver Yin and kidney Yin and deficiency of both qi and yin,patients with deficiency of Yang in the spleen and kidney exhibit a significantly increased peripheral blood Th17/Treg ratio (P<0.05), there is no statistically significant difference between the other patients with principle syndrome(P>0.05).(8)When it comes to TCM accompanied symptoms and signs, the Th17cells proportion of patients with upper hyperactivity of liver yang, blood stasis, retained fluid and turbid toxin and is higher than that in patients without accompanied symptoms and signs (P<0.05), and the patients with turbid toxin showed most obviously,patients with upper hyperactivity of liver yang,blood stasis syndrone type, retained fluid and dampness-heat appear resemblance, there is also no obviously difference between patients with dryness-heat and dampness-heat(P>0.05).(9)Compared with patients without accompanied symptoms and signs, patients with upper hyperactivity of liver yang, blood stasis,dryness-heat and turbid toxin have a significant reduction of the proportion of Treg cells (P<0.05), patients with blood stasis show a significantly decreasing Treg cells percentage compared with patients with retained fluid (P<0.05),the changes among patients with other accompanied symptoms and signs are not significantly different(P>0.05).(10)The level of Th17/Treg ratio among patients with blood stasis, dampness-heat, retained fluid and turbid toxin presents is no statistically different (P>0.05), but significantly elevates in comparison with patients with upper hyperactivity of liver yang and patients without accompanied symptoms and signs (P<0.05), Th17/Treg ratio of patients with turbid toxin and blood stasis is higher than dryness-heat patients (P<0.05), there is no statistical significance among patients with other TCM accompanied symptoms and signs (P>0.05).ConclusionThl7/Treg imbalance exists in DN patients.The proportion of Th17cells of peripheral blood and plasma IL-17A are elevated,while the proportion of Treg cells is decreased.Interestingly,the Treg cells related cytokines IL-10and TGF-betal are increased.However,the ratio of Th17/Treg is raised along with the progression of the disease.The more urine proteinse is eccrisised,the more obvious these changes are,suggesting that the imbalance of Th17and Treg cells is involved in DN and the ratio of Th17/Treg cells,index of the imbalance of Th17and Treg cells, can more credibly reveal immune inflammation mechanism of DN. The imbalance of Th17/Treg of patients with deficiency of Yang in the spleen and kidney and dampness toxin, blood stasis, dampness-heat,retained fluid syndrome performs more distinctly,which provides objective evidence for us to treat DN based on syndrome differentiation,and helps us search new targets to prevent and cure DN with TCM. |
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