| Objective: This study combines biochemical and pathological indicators with metabonomics technology, discusses the mechanism of action of stilbene art capsule treatment of DN,and perfect the evaluation system of the stilbene capsule in the treatment of DN curative effect. At the same time, the study uses microscopic indexes in combination with the theory of traditional Chinese medicine, applies the metabonomics technology demonstration by taking stilbene art capsule spleen, and activating blood heat, dry wet treatment the curative effect of DN, provide new idea and method of controling the DN. Methods: Add doses(50 mg/kg) of disposable injection chain urea with cephalosporins(STZ) to ordinary feed to induce DN rat model for 8 weeks.Dicide the normal and building successful rats(according to random number table method) into four groups: normal group, stilbene capsule group, the cozaar group, model group, 10 in each group. After 8 weeks, 1) use automatic biochemical analyzer to test glycosylated hemoglobin(HbAlc), renal urea nitrogen(BUN), creatinine(Scr), serum(inhibition(CysC), 24 hours urinary protein quantitative(24hpro), c-reactive protein(CRP); 2) use enzyme-linked immunoassay ELISA method to detect serum FN, MMP2 expression; 3) observe the development of glomerular disease and renal tubular interstitial fibrosis,under the light microscope of kidney tissues HE staining; 4) use nuclear magnetic resonance hydrogen spectrum(1h-nmr), least squares criterion(PLS-DA) and orthogonal partial least squares discriminant analysis(OPLS-DA) methods to analyze the normal group, model group, cozaar group and stilbene capsule on DN rats serum and kidney tissue metabolism, find out the DN metabolic marker,and explore stilbene metabolites trajectory after capsule treatment of DN. Results:(1)The effect of stilbene capsule on glycosylated hemoglobin(HbAlc) and the renal function.Compared with normal group, model group rats HbAlc content significantly increased(P < 0.01). Compared with model, stilbene capsule group showed decrease, and the art of stilbene capsule group(P < 0.05) showed that the stilbene capsule can significantly reduce the DN rat hemoglobin a1 c levels.Cozaar had no effect.Compared with the normal group, serum BUN, Scr, Cys-c and 24 h pro of model group rats increased significantly(P < 0.01); Compared with model group, stilbene capsule(P < 0.01), cozaar(P < 0.05) decreased, which indicates that(dose) stilbene capsule not only can reduce rats serum BUN, Scr and Cys-c and 24 h pro role, but also can improve renal function which is superior to western medicine treatment group(low dose).(2)The effect of stilbene capsule on the change of kidney pathological morphologyUnder the light microscope of HE staining: normal group of glomerular structure was integrate without obvious inflammatory cells infiltration; Glomerular basement membrane of model group became thicker diffusely, mesangial in a mild-to-moderate degree thickened, Capillary tumor samples charged, Renal tubular expanded, renal interstitial inflammatory cells and lymphocyte infiltrated; In stilbene capsule group and cozaar, there were similar kidney pathological changes, but they were less to some degree in the model group(P < 0.05). All these indicate that stilbene art capsule can delay the progress of DN renal tissue fibrosis.(3)The changes of stilbene capsule in the treatment of DN metabolomics1)The adjustment of stilbene capsule on DN kidney metabonomicsCompared with normal group,model group has nine endogenous metabolites which show significant difference(P < 0.05),among which betaine, creatinine, adenosine, formic acid, nicotinamide,and taurine content are higher,while the content of tyrosine, tryptophan glutamic acid(P < 0.05) goes down. Compared with model group, stilbene capsule can readjust these 9 different ingredients to some degree; In cozaar group,can readjust,it can adjust more or less 5 different ingredients,which indicate that stilbene capsule is more effective in treating of diabetic nephropathy.2)The adjustment of stilbene capsule on the regulation of serum metabolomics DNCompared with normal group, model group contained six endogenous metabolites which showed significant difference(P < 0.05), among which the content of betaine, isoleucine, and 3- HB, acetone, succinic acid, tyrosine went down(P < 0.05). Compared with model group, stilbene capsule could readjust 5 different ingredients to some degree.In Cozaar group,four ingredients could be adjusted,but the adjustment trend was not alway biased towards the normal group.The result showed that stilbene capsule was more effective in treating diabetic nephropathy. Conclusion:This study showed that stilbene art capsule could prevent its occurrence and development in its early,mid period by combining biochemical and pathological indicators with metabonomics technology,and by asessing comprehensive curative effect. Through the application of metabonomics technology, 9 mid kidney tissues species, 6 kinds of serum endogenous metabolites could be identified in the earlyand mid process of DN.After drug intervention, these markers varied to some extent(P < 0.05), and stilbene is better than that of cozaar group(P < 0.05).The study paved the way for further exploration of the pathogenesis of DN, early diagnosis and the evaluation of curative effect. |
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