AMPK-mediated Anti-DKD Effect Of Small Molecule Compound X22

Diabetic Kideney Disease(DKD)is one of the common complications in diabetic patients and the main cause of end-stage renal disease(ESRD).Its pathogenesis is related to abnormal glucose metabolism,inflammatory response,oxidative stress and so on.At present,there is still no specific drug for the treatment of DKD,and patients can only delay the deterioration of their disease by taking hypoglycemic drugs for a long time,resulting in a decline in patients’ quality of life and a huge economic burden.Therefore,it is of great significance to study the treatment of DKD.As an energy regulator,adenosine-activated protein kinase(AMPK)not only plays an active role in glucose and lipid metabolism,but also has anti-inflammatory and antioxidant effects.Studies have shown that its decreased activity is closely related to the development of DKD,and AMPK is expected to become an important drug target for the treatment of DKD.Small molecule compound X22 is a novel pyridine-imidazole derivative,whose parent nuclear pyridine-imidazole can directly activate AMPK,and X22 also has a good therapeutic effect on DKD,but the mechanism of action is still unclear.In this study,streptozotocin(STZ)-induced DKD rat model and high glucose-induced mesangal cell(MS-13)proliferation model were constructed to investigate whether X22 exerts anti-DKD renal injury through activation of AMPK.(1)The degree of glomerular lesion is still an important criterion for DKD classification.DKD rat model was established by a single injection of 60mg/kg STZ into the tail vein and induced by the development of the disease for 12 weeks.DKD rat model was administered for 8 consecutive weeks(10m L/kg intragastric administration of corresponding dose of liquid,once a day).The experimental results showed that glomerular lesions such as glomerular hypertrophy,mesangial matrix hyperplasia and glomerular basement membrane thickening were found in the renal tissues of rats in model control group.X22 significantly improved glomerular hypertrophy,mesangial matrix hyperplasia,glomerular basement membrane thickening and other pathological changes,significantly increased urinary creatinine clearance,restored the content of urea nitrogen(BUN),significantly upregulated AMPK protein expression level.It inhibited the expression levels of transforming growth factor-β1(TGF-β1),connective tissue growth factor(CTGF)and α smooth muscle actin(α-SMA).(2)The proliferation of mesangial cells is an early pathological change of DKD.The experiment used m ES-13 proliferation induced by high glucose as a model.The results showed that mesangial cells(MS-13)proliferated significantly under high glucose induction.X22 inhibited proliferation at 24,48 and 72 h.AMPK inhibitors reversed the inhibitory effect of X22,suggesting that the inhibitory effect of X22 on the proliferation of mesangial cells induced by high glucose was related to AMPK.Further Western Blot results showed that X22 inhibited the proliferation of mesangial cells(MS-13)induced by high glucose by phosphorylation of AMPK.In this study,we found that X22 can activate AMPK and inhibit TGF-β1/CTGF pathway to play an anti-DKD renal injury effect,which is expected to provide new ideas for drug research and development of DKD patients.