1.Effect and mechanism of Ginkgo biloba extract on acute lung injury induced by sepsis in mice
Xiaoshua WEI ; Yating WEI ; Hua XIAO ; Chaoqian LI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2025;32(4):404-409
Objective To investigate the effect and mechanism of Ginkgo biloba extract(EGB761)on acute lung injury(ALI)induced by sepsis in mice.Methods Fifty male Kunming mice were selected,they were randomly divided into sham operation group(intraperitoneal injection of equal amount of normal saline after operation),model group(intraperitoneal injection of equal amount of normal saline after operation),low-dose EGB761 group(intraperitoneal injection of 20 mg/kg EGB761 immediately after operation)and high-dose EGB761 group(intraperitoneal injection of 50 mg/kg EGB761 immediately after operation)and high-dose EGB761+nuclear factor E2-related factor 2(Nrf2)inhibitor ML385 group(intraperitoneal injection of ML385 30 mg/kg 1 hour before modeling and 50 mg/kg EGB761 immediately after operation),with 10 mice in each group.ALI model of sepsis was established by cecal ligation and puncture(CLP),and the sham operation group only underwent open-closed abdominal surgery.Serum and lung tissue were taken at 12 hours after operation,and the pathological changes of lung tissue were observed under light microscope after hematoxylin-eosin(HE)staining,and the lung injury score was made.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin(IL-6,IL-1β)and tumor necrosis factor-α(TNF-α)in serum of mice in each group.Western blotting test and real-time fluorescence quantitative reverse transcription-polymerase chain reaction(qRT-PCR)were used to detect the protein expression levels of glutathione peroxidase 4(GPX4),Nrf2,member 11 of solute carrier family 7(SLC7A11)and the mRNA expression levels of Nrf2 and GPX4 mRNA the lung tissues of mice in each group.The content of malondialdehyde(MDA)in the lung tissues of mice in each group was detected by the thiobarbituric acid method.The activity of superoxide dismutase(SOD)in the lung tissues of mice in each group was detected by xanthine oxidase method.Results Compared with the sham operation group,in the model group,a large number of inflammatory cells infiltrated in the lung tissue,the alveolar structure was destroyed,the lung injury score was significantly increased(3.71±0.29 vs.0.33±0.10),and serum pro-inflammatory factors IL-1β,IL-6,TNF-α and the content of oxidation index MDA in lung tissue all increased significantly[IL-1β(ng/L):0.410±0.002 vs.0.351±0.007,IL-6(ng/L):0.650±0.010 vs.0.560±0.010,TNF-α(ng/L):0.320±0.010 vs.0.290±0.002,MDA(nmol/mg):2.49±0.31 vs.1.77±0.02,all P<0.05],while SOD activity in lung tissue decreased significantly(U/mg:20.20±4.61 vs.61.47±8.10,P<0.05),the expression levels of Nrf2,GPX4,SLC7A11 protein,GPX4,and Nrf2 mRNA all decreased[NRF2 protein expression(Nrf2/β-actin):0.33±0.11 vs.0.63±0.17,GPX4 protein expression(GPX4/β-actin):1.19±0.08 vs.2.50±0.17,SLC7A11 protein expression(SLC7A11/β-actin):0.41±0.03 vs.0.67±0.08;GPX4 mRNA(2-ΔΔCt):0.32±0.00 vs.1.02±0.21,NRF2 mRNA(2-ΔΔCt):0.44±0.04 vs.1.01±0.20,all P<0.05].Compared with the model group,the degree of lung injury in the low-dose EGB761 group and the high-dose EGB761 group was reduced to varying degrees,and the lung injury score in the high-dose EGB761 group was significantly reduced(0.96±0.40 vs.3.71±0.29,P<0.05).The contents of MDA in lung tissue and serum proinflammatory factors IL-1β,IL-6 and TNF-α were significantly decreased[MDA(nmol/mg):1.53±0.09 vs.2.49±0.31,IL-1β(ng/L):0.350±0.004 vs.0.410±0.002,TNF-α(ng/L):0.280±0.003 vs.0.320±0.010,all P<0.05],SOD activity,proteins of Nrf2,GPX4 and SLC7A11,and expression levels of GPX4 and Nrf2 mRNA all increased significantly[SOD activity(U/mg):63.78±8.74 vs.20.20±4.61,the expression of Nrf2 protein(NRF2/β-actin):0.71±0.06 vs.0.33±0.11,the expression of GPX4 protein(GPX4/β-actin):2.03±0.10 vs.1.19±0.08,and the expression of SLC7A11 protein(SLC7A11/β-actin)0.65±0.07 vs.0.41±0.03;GPX4 mRNA(2-ΔΔCt):0.80±0.10 vs.0.32±0.00,NRF2 mRNA(2-ΔΔCt):0.84±0.07 vs.0.44±0.04,all P<0.05].Compared with high-dose EGB761 group,Nrf2 inhibitor ML385 can obviously reverse the above-mentioned effects of high-dose EGB761 after intervention.Conclusion Ginkgo biloba extract EGB761 can improve the sepsis ALI induced by CLP in mice,and it may play a protective role by activating Nrf2 and up-regulating SLC7A11 to inhibit ferroptosis.
2.Effect and mechanism of Ginkgo biloba extract on acute lung injury induced by sepsis in mice
Xiaoshua WEI ; Yating WEI ; Hua XIAO ; Chaoqian LI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2025;32(4):404-409
Objective To investigate the effect and mechanism of Ginkgo biloba extract(EGB761)on acute lung injury(ALI)induced by sepsis in mice.Methods Fifty male Kunming mice were selected,they were randomly divided into sham operation group(intraperitoneal injection of equal amount of normal saline after operation),model group(intraperitoneal injection of equal amount of normal saline after operation),low-dose EGB761 group(intraperitoneal injection of 20 mg/kg EGB761 immediately after operation)and high-dose EGB761 group(intraperitoneal injection of 50 mg/kg EGB761 immediately after operation)and high-dose EGB761+nuclear factor E2-related factor 2(Nrf2)inhibitor ML385 group(intraperitoneal injection of ML385 30 mg/kg 1 hour before modeling and 50 mg/kg EGB761 immediately after operation),with 10 mice in each group.ALI model of sepsis was established by cecal ligation and puncture(CLP),and the sham operation group only underwent open-closed abdominal surgery.Serum and lung tissue were taken at 12 hours after operation,and the pathological changes of lung tissue were observed under light microscope after hematoxylin-eosin(HE)staining,and the lung injury score was made.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin(IL-6,IL-1β)and tumor necrosis factor-α(TNF-α)in serum of mice in each group.Western blotting test and real-time fluorescence quantitative reverse transcription-polymerase chain reaction(qRT-PCR)were used to detect the protein expression levels of glutathione peroxidase 4(GPX4),Nrf2,member 11 of solute carrier family 7(SLC7A11)and the mRNA expression levels of Nrf2 and GPX4 mRNA the lung tissues of mice in each group.The content of malondialdehyde(MDA)in the lung tissues of mice in each group was detected by the thiobarbituric acid method.The activity of superoxide dismutase(SOD)in the lung tissues of mice in each group was detected by xanthine oxidase method.Results Compared with the sham operation group,in the model group,a large number of inflammatory cells infiltrated in the lung tissue,the alveolar structure was destroyed,the lung injury score was significantly increased(3.71±0.29 vs.0.33±0.10),and serum pro-inflammatory factors IL-1β,IL-6,TNF-α and the content of oxidation index MDA in lung tissue all increased significantly[IL-1β(ng/L):0.410±0.002 vs.0.351±0.007,IL-6(ng/L):0.650±0.010 vs.0.560±0.010,TNF-α(ng/L):0.320±0.010 vs.0.290±0.002,MDA(nmol/mg):2.49±0.31 vs.1.77±0.02,all P<0.05],while SOD activity in lung tissue decreased significantly(U/mg:20.20±4.61 vs.61.47±8.10,P<0.05),the expression levels of Nrf2,GPX4,SLC7A11 protein,GPX4,and Nrf2 mRNA all decreased[NRF2 protein expression(Nrf2/β-actin):0.33±0.11 vs.0.63±0.17,GPX4 protein expression(GPX4/β-actin):1.19±0.08 vs.2.50±0.17,SLC7A11 protein expression(SLC7A11/β-actin):0.41±0.03 vs.0.67±0.08;GPX4 mRNA(2-ΔΔCt):0.32±0.00 vs.1.02±0.21,NRF2 mRNA(2-ΔΔCt):0.44±0.04 vs.1.01±0.20,all P<0.05].Compared with the model group,the degree of lung injury in the low-dose EGB761 group and the high-dose EGB761 group was reduced to varying degrees,and the lung injury score in the high-dose EGB761 group was significantly reduced(0.96±0.40 vs.3.71±0.29,P<0.05).The contents of MDA in lung tissue and serum proinflammatory factors IL-1β,IL-6 and TNF-α were significantly decreased[MDA(nmol/mg):1.53±0.09 vs.2.49±0.31,IL-1β(ng/L):0.350±0.004 vs.0.410±0.002,TNF-α(ng/L):0.280±0.003 vs.0.320±0.010,all P<0.05],SOD activity,proteins of Nrf2,GPX4 and SLC7A11,and expression levels of GPX4 and Nrf2 mRNA all increased significantly[SOD activity(U/mg):63.78±8.74 vs.20.20±4.61,the expression of Nrf2 protein(NRF2/β-actin):0.71±0.06 vs.0.33±0.11,the expression of GPX4 protein(GPX4/β-actin):2.03±0.10 vs.1.19±0.08,and the expression of SLC7A11 protein(SLC7A11/β-actin)0.65±0.07 vs.0.41±0.03;GPX4 mRNA(2-ΔΔCt):0.80±0.10 vs.0.32±0.00,NRF2 mRNA(2-ΔΔCt):0.84±0.07 vs.0.44±0.04,all P<0.05].Compared with high-dose EGB761 group,Nrf2 inhibitor ML385 can obviously reverse the above-mentioned effects of high-dose EGB761 after intervention.Conclusion Ginkgo biloba extract EGB761 can improve the sepsis ALI induced by CLP in mice,and it may play a protective role by activating Nrf2 and up-regulating SLC7A11 to inhibit ferroptosis.
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