Inhibition Of GLS1 Ameliorates Aging-related Diseaseses

Objective:In this study,glutaminase inhibitors（BPTES）were used to treat the aging-related diseases chronic heart failure and liver fibrosis,respectively.To explore whether inhibition of glutaminase 1（GLS1）can improve heart failure induced by pressure overload and reduce carbon tetrachloride induced liver fibrosis in mice.Methods:1.Fibroblasts were treated with DNA-damaging agent doxorubicin（DOX）or transforming growth factor-β（TGF-β）for 48 hours to construct a senescence model.SA-β-Gal staining was used to detect cell senescence.The expression of senescence markers（P16,P21）,senescence-related secretory phenotypes（IL-1b,IL-6,CCL2）and glutaminase 1（GLS1）at m RNA level were detected by q RT-PCR.Cell proliferation was detected by Brdu+staining.Senescent fibroblasts were treated with glutaminase inhibitors（BPTES）.Cell viability assays were used to evaluate the killing ability of BPTES against senescent cells.2.Transverse aortic constriction（TAC）was used to simulate pressure overload-induced heart failure in mice to establish a chronic heart failure model.The blood flow velocity and cardiac function at the aortic ligation site after TAC were detected by small animal ultrasound to determine the success of TAC operation.SA-β-Gal staining was used to observe the senescence of heart tissue.The collagen deposition was observed by Sirius red staining.3.The mouse model of liver fibrosis was established by intraperitoneal injection of olive oil（control group,N=10）,10%CCl4（10μL/g）（model group,N=10）or 10%CCl4（10μL/g）+BPTES（10mg/kg）（treatment group,N=10）for four weeks.Sirius red staining was used to observe the collagen deposition in liver tissue.Flow cytometry was used to detect the infiltration of various inflammatory cells in liver tissue.q RT-PCR and immunohistochemical staining were used to detect the expression of P16,a cellular senescence marker in mouse liver tissue.The m RNA levels of senescence-related secretory phenotype-related proteins（IL-6,CCL2,CXCL1）,fibrosis-related genes（Acta2,Col1a1）and glutaminase 1（GLS1）were detected by q RT-PCR.The expression of GLS1was detected by immunohistochemistry.Results:1.Cell viability assays showed that senescent cell death was significantly induced in the BPTES treatment group compared with the DMSO group.2.Compared with the control group,the heart volume and weight of the model group increased.Small animal ultrasound showed that the blood flow velocity increased and the heart function decreased at the ligation site of the aortic arch in the model group.Sirius red staining showed increased myocardial fibrosis in the model group.Compared with the model group,the heart volume and weight of the BPTES treatment group decreased.Echocardiography showed that the cardiac function of the treatment group was improved.Sirius red staining showed that the degree of cardiac fibrosis was reduced in the BPTES treatment group.3.Compared with the control group,the liver tissue of the model group was generally enlarged,the surface was not smooth and granular,and the ratio of liver weight to tibia length was significantly increased.Sirius red staining showed that collagen deposition in the liver tissue of the model group was significantly increased.q RT-PCR results showed that the expressions of fibrosis-related genes（Acta2 and Col1a1）and GLS1 were significantly up-regulated in liver tissues.The results of immunohistochemical staining showed a significant increase in GLS1 levels.Compared with the model group,the liver weight of the BPTES treatment group was significantly reduced.The area of collagen deposition with Sirius red staining positive was significantly reduced.q RT-PCR results showed that the expression of fibrosis-related genes Acta2 and Col1a1 in the liver tissues was significantly decreased in the BPTES treatment groupConclusion:1.Inhibition of glutaminase 1 activity can improve cardiac function and myocardial fibrosis in TAC-induced heart failure,which provides a new idea for the treatment of chronic heart failure.2.Inhibition of glutaminase 1 activity can significantly improve CCL₄-induced liver fibrosis,providing a new idea for the treatment of liver fibrosis.