This study aims to explore the effects and mechanisms of the core formula(TCF) improving glomerulosclerosis(GS) in diabetic kidney disease(DKD) by Jinling medical school based on reticulophagy(ER-phagy)-pyroptosis-apoptosis-necroptosis(PANoptosis) signaling pathway in podocytes. Firstly, the modified DKD rat models were established. After modeling successfully, the DKD rat models received either TCF, empagliflozin(EMPA), or saline by gavage for 8 weeks, respectively. Secondly, MPC5 cells were subjected to treatment under high-glucose(HG) conditions alone, or HG combined individually with drug-containing serum of the low-dose of TCF(L-TCF) and the high-dose of TCF(H-TCF), EMPA, or Torin 1(autophagy agonist). In the in vivo studies, the changes of general conditions and renal injury indicators, pathological characteristics of GS, and the protein expression levels of podocyte marker molecules were observed, respectively. Furthermore, the changes of the level of reactive oxygen species(ROS) in the kidneys, the expression levels of the core receptor and marker molecules of ER-phagy, and the expression degree of Z-DNA binding protein 1(ZBP1) were also observed. In the in vivo and in vitro studies, we analyzed changes in the expression levels of ZBP1 protein in the kidneys and podocytes, as well as the protein expression levels of key signaling molecules in the cellular PANoptosis were investigated, respectively. The in vivo study results showed that, for the DKD model rats, general conditions and renal injury indicators, pathological characteristics of GS, and the protein expression levels of podocyte marker molecules were improved, respectively. Moreover, the level of ROS in the kidneys, the expression level of the core receptor and marker molecules of ER-phagy, and the expression degree and protein expression level of ZBP1, as well as the protein expression levels of key signaling molecules in the cellular PANoptosis were ameliorated, respectively. Notably, the improvement effects of TCF and EMPA were basically similar, but the beneficial effect of the protein expression level of interleukin-18(IL-18) in the kidneys, as an inflammatory factor, was superior to EMPA. The in vitro study results showed that, high and low doses of TCF and EMPA both could ameliorate the protein expression levels of key signaling molecules in the PANoptosis pathway in podocytes treated with HG, and the improvement effect of H-TCF was superior to that of L-TCF or EMPA. In conclusion, the study clarifies that TCF improves GS in DKD by regulating ER-phagy-PANoptosis signaling pathway in podocytes in vivo and in vitro, and has laid the groundwork for exploring mechanisms and scientific implications of treating diabetic kidney disease by Jinling medical school.
