Transcriptomics and Proteomics Reveal That TLPW Acupuncture Ameliorates Proteinuria in Diabetic Kidney Disease Model Rats by Suppressing Epithelial-to-Mesenchymal Transition via the DPP4/SDF-1α/TGF-β/Smad Signalling Axis.

Abstract

Ten male Sprague-Dawley rats were randomly assigned to serve as the negative control (NC) group, and 50 others were maintained on a high-fat diet (HFD). After 18 weeks of feeding, the HFD group received streptozotocin (STZ) intraperitoneally to establish a DKD model. HFD group rats (24-h urinary protein excretion rate ≥ 30 mg) were randomly divided into model (DKD), tiaolipiwei acupuncture (DKD + Acu, acupuncture for 30 min), AMD3100 (DKD + AMD3100, AMD3100 intraperitoneal injection) and tiaolipiwei acupuncture + AMD3100 group (DKD + Acu + AMD3100, AMD3100 intraperitoneal injection combined with tiaolipiwei acupuncture for 30 min) groups. The intervention lasted 4 weeks, and body weight and random blood glucose levels were recorded for each group before treatment each week. Postintervention (at 4 weeks), urine was collected to assess urinary protein-creatinine ratios, 24-h urinary protein contents and urinary podocyte injury-related enzyme levels. Renal cortex tissues from three to five rats in the control, DKD and DKD + Acu groups were sent for transcriptomic and proteomic analyses, and renal tissues were collected for analyses of pathological indicators and mechanisms. Twenty-four-hour Upro, 24-h urinary Spondin 2 (SPON2) levels, UACRs and random serum creatinine, urea nitrogen and blood glucose levels in the DKD + Acu group exhibited significantly reduced levels compared to the DKD group (#p < 0.05). According to the transcriptomic 2.2.2s and proteomic results, immunofluorescence or western blotting was used to assess podocyte-specific marker expression levels (nephrin, podocin and CD2AP), epithelial-mesenchymal transition (EMT) markers (desmin, Fsp1 and α-SMA) and DPP-4/SDF-1α/TGF-β/Smad signalling axis components; nephrin, podocin and CD2AP expression significantly elevated (#p < 0.05 or ##p < 0.01) and desmin, Fsp1 and α-SMA expression greatly decreased (#p < 0.05 or ##p < 0.01) in the DKD + Acu group. Tiaolipiwei acupuncture regulated the Dpp-4/SDF-1α/TGF-β/Smad signalling axis (#p < 0.05 or ##p < 0.01), but this effect was reduced by AMD3100 (@p < 0.05 or @@p < 0.01). Tiaolipiwei acupuncture modulates the DPP-4/SDF-1α/TGF-β/Smad signalling axis to inhibit podocyte EMT and alleviate podocyte and renal injury, ultimately ameliorating proteinuria in DKD model rats.