Clinical Research On Pathogenesis Of Internal Heat In Diabetic Nephropathy And Research On The Intervention Mechanism Of "clearing Heat And Eliminating Symptoms"

With worldwide epidemic of diabetes mellitus(DM),diabetic kidney disease(DKD)has become the primary cause of Chronic Kidney Disease(ESRD)in China.Over time,the current treatment for DKD are not adequate and lack of breakthrough.The Chinese medicine formula show its superiority by polypharmacology and network pharmacology to produce an overall systemic effect.The etiology of DKD in TCM theory need to be further explored and enriched.Pre viously the core pathogenesis of DM was "Yin Deficiency-Dryness Heat",however the mechanism has changed because of the modern lifestyle.As the theory "Superheat Consuming Qi" showed by Huangdi medical classic,the Heat is important in the development of DKD.We proposed the hypothesis that "the renal collateral conglomeration induced by superheat" along with "Heat and Conglomeration Clearing Method"based on our previous "renal collateral micro-conglomeration"theory.Moreover,the renal protective effect of "Qing Re Xiao Zheng Formula"(QRXZF)which was summarized for the treatment of DKD early and middle stage maybe associated with the inhibition of renal micro-inflammation.Objective:1.To investigate the distribution of Heat Syndrome and correlation with renal function in DKD patients in different stages.2.To explore the effect of QRXZF on rat mensaginal cells stimulated by highglucose through the p38MAPK signal pathway.Methods:1.The clinical and TCM syndrome information of 335 DKD patients were collected and analyzed by t test,non-parametric test,stepwise regression analysis with spss statistical software.2.The rat Mesangial Cells(MCs)were stimulated by highglucose and the Serum with QRXZF or Irbesatan was applied to the mesangial cells to test the effect on cell proliferation,hypertrophy,inflammatory cytokines and Fibrosis factors excretion as well as p38MAPK signaling pathway activation.Results:(1)Elderly people with overweight or obesity were the majority of DKD patients,Heat Syndrome was widespread in the whole DKD course,its widely combination with Qi and Yin Difficiency Syndrome proved the traditional Chinese medicine theory "Superheat Consuming Qi "and" Superheat Consuming Yin".(2)Compared with non-Heat Syndrome group,The eGFR in Heat Syndrome group was higher in the early stage;the 24h proteinuria quantification was higher in the middle stage;the Scr and Bun is higher but the Eco2 and eGFR is lower in the late stage.(3)There was correlation of HbAlc and the Heat Integration in the early stage proved by the stepwise regression analyzed equation [y(HeatIntegration)=6.616+0.548x1(HbA1C)+0.246X2(Yin-Difficiencylntegration);y(HbA1c)=5.750+0.208x1(FBG)+0.099x2(Heat Integration)](4)The eGFR was showed positively correlated with the Heat Integration in early stage as showed by the stepwise regression analyzed equation[y(eGFR)=83.007+2.153x(Heat Integration)];Interestingly,the eGFR was negatively correlated with the Heat Integration while the Heat Integration had positive correlation with Dam-Turbidity Integration but negative correlation with eGFR and Qi Difficiency Integration [y(Heat Integration)=13.91+0.596x1(Damp-Turbidity Integration)-0.573X2(Qi-Difficiency Integration)-0.206X3(eGFR);y(eGFR)=25.347-0.501 X(Heat Integration)](5)Compared with the people without diabetics,the serum p-p38MAPK in DKD patients was high expressed,and haspositivecorrelation with Heat Intergration[y(p-p38MAPK)=9.499+0.889x(内热积分)](6)Highglucose promoted the proliferation and hypertrophy of mesangial cells,especially in 48h and 30mmol/L concentration.The p38MAPK/CREB signal pathway was activated along with the excessive excretion of MCP-1、TNF-α、IL-6、FN and Collagen Ⅳ in highglucose stimulation condition.The excessive secretion of ECM in rat MCs mediated by highglucose might be caused by both cell proliferation and cell hypertrophy.(7)Compared with highglucose group,the cell proliferation and hypertrophy was inhibited in QRXZF group,moreover the phosphoralation of P38MAPK/CREB and excretion of MCP-1、FN was decreased showed by western-blot and PCR test.The holistic protective effect of QRXZF was much better than Irbesatan.Conclution:1.Heat Syndrome which accentuated the renal function was common in the whole DKD course,highglucose and overweight were the foundation to heat generation,it would accelerate eGFR to induce high perfusion,high filtration in early-stage and aggravated the albuminuria in the middle stage,at last deteriorated the decreation of eGFR in the late-stage.2.QRXZF ameliorated the proliferation and hypertrophy of MCs induced by highglucose,meanwhile the phosphoralation of p38MAPK/CREB and MCP-1,FN expression were inhibited.The inhibition of p38MAPK signal pathway maybe one of the complex intervention mechanisms of QRXZF.3.The rules of Heat Clearing in clinical treatment for DKD would be applied in clinic according to this study.Clearing the heat hided behind the Qi-Yin deficiency was the primary principle in the early stage,then nourishing Qi and yin.Diathermy method including Heat Clearing,Conglomeration Dissipating was important to prevent the stagnation of conglomeration in middle-stage,in addition,Dissolving damp-turbidity and heat was important in the late stage as well asreplenishing Qi and immunity.