Efects Of Emodin On The Expression Of Intergern Linked Kinase In Podocyte Epithelial-mesenchymal Transition In The Early Stage Of DKD

Object1. Observe the expression change of podocyte polarity markers desmin and nephrin, investigate the effects of high glucose(HG) on podocyte epithelial-to-mesenchymal transition(EMT) in-vitro and in-vivo.2. Observe the expression change of integrin linked kinase(ILK) in the process of podocyte EMT, investigate the role of ILK signaling way in podocyte EMT.3. Observe the effects of traditional Chinese medicine monomer emodin on podocyte EMT and expression of ILK, investigate the mechanism of emodin in the treatment of diabetic kidney disease(DKD).MethodIn-vitro studiesDitionally immortalized mouse podocytes were divided into the following group:(1) normal glucose group(NG) as control incubated in RPMI 1640 containing 5m M glucose;(2) high glucose group(HG) incubated in RPMI 1640 medium containing 30 mM glucose;(3) emodin group(EMO) incubated in HG medium supplemented with emodin(30ug/ml).(4) ILK blocking group incubated in HG medium supplemented with ILK inhibitor QLT0267. ILK, nephrin and desmin expression change were analized by western blot, real time rt-PCR and immunofluorescence, spearman correlation analysis analy the correlation between indicators.Animal studiesStreptozotocin(STZ)-induced diabetic rats were randomly divided into the following groups:(1) the DM group: treated with the same volume of saline;(2) DM + emodin group: treated with a dose of EMO at 20 mg·kg-1·d-1;(3) DM + ILK inhibitor: treated with a dose of ILK inhibitor, QLT0267 at 10 mg·kg-1.w-1;(4) normal control group, Normal Sprague-Dawley rats were choosed. At the end of the 12-week treatment, animals were then killed and the kidneys were harvested immediately. ILK, nephrin and desmin expression change were analized by western blot, real time rt-PCR and immunohistochemistry. spearman correlation analysis analy the correlation between indicators, biochemical indexes and proteinuria.Results1. Compared with NG, podocyte exposured to HG resulted in a remarkable increase of desmin, decrease of nephrin, high glucose induced podocyte EMT; Compared with normal rat, in the renal tissue from diabetic rats the nephrin expression was reduced and the desmin expression was increase, urinary ACR significantly increased.2. The ILK expression increased in the high glucose induced podocyte EMT, in the renal tissue from diabetic rats the ILK expression was increase, ILK inhibitor reduced the podocyte EMT, reduced urinary ACR.3. Emodin reduced the high glucose induced podocyte EMT and decreased ILK expression; emodin partially abrogated diabetic rats podocyte EMT, reduced urinary ACR.Conclusion1. High glucose can induce podocyte EMT, diabetic rats renal tissue present podocyte EMT and urinary ACR increase.2. High glucose induced podocyte EMT by adjusting the ILK signaling pathways.3. Emodin by blocking ILK signaling pathway to restrain high glucose induced podocyte EMT, reduce proteinuria.