Abstract:

Objective To investigate the role of lncRNA SNHG1 in homocysteine-induced pyroptosis of podocyte. Methods Cbs ^+/- mice were randomly divided into two groups： a normal diet group （ND） and a high methionine diet group （HMD）. Western blotting was used to detect the protein expression levels of Caspase-1， Cleaved Caspase-1， and NLRP3. Mouse renal glomerular podocytes were cultured in vitro， and then assigned into a control group （Control， 0 μmol/L Hcy） and a homocysteine intervention group（Hcy， 80 μmol/L Hcy）. Western blotting was used to detect the protein expression levels of Caspase-1， Cleaved Caspase-1， and NLRP3. Mouse renal glomerular podocyion group （OE-NC + Hcy） and the lncSNHG1 overexpression + homocysteine intervention group （OE-SNHG1 + Hcy） were also established.After 48 hours of intervention， Real-time fluorescence quantitative PCR was used to detect the expression of lncSNHG1 in podocytes after Hcy intervention. Western blot was used to detect the expressions of Caspase-1，Cleaved Caspase-3 and NLRP3. Immunofluorescence was used to detect the expression levels of GSDMD and GSDMD-N. ELISA was used to detect the contents of IL-1β and IL-18. Results （1）In the animal experiments， the expression levels of pyroptosis-related proteins Caspase-1， Cleaved Caspase-1，NLRP3， GSDMD， and GSDMD-N were all increased in the HMD group compared with the ND group. （2）In the cellular experiments， the expression levels of Caspase-1，Cleaved Caspase-1，NLRP3， GSDMD， and GSDMD-N were all increased in the Hcy group compared with the Control group， and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased as well. （3） In the cellular experiments， the expression of lncSNHG1 was increased in the Hcy group compared with the control group. After transduction with lncSNHG1 lentivirus， the expression of lncSNHG1 was increased in the OE-SNHG1 group，compared with the control group and the OE-NC group. （4）In the cellular experiments， the expressions of pyroptosis-related proteins Caspase-1，Cleaved Caspase-1， NLRP3， GSDMD，and GSDMD-N were increased compared with the OE-NC+Hcy group， and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased in the OE-SNHG1+Hcy group. Conclusion These results indicate that lncSNHG1 may play a role in promoting Hcy induced podocytepyroptosis.

Key words: homocysteine, podocyte, small nucleolar RNA host gene 1, pyroptosis