Interventional effect of total flavonoids of Ginkgo biloba on human embryonic kidney cytotoxicity induced by mercury poisoning and its effect on SIRT1/NLRP3 pathway

Abstract

Abstract: Objective To investigate the intervention effect of total flavonoids of Ginkgo biloba on the cytotoxicity of human embryonic kidney induced by mercury poisoning and its effect on silent information regulator 1（SIRT1）/nod-like receptors-3（NLRP3） pathway. Methods HEK293 cells were cultured in vitro and included the control group,mercury poisoning model group（1 000 μmol/L HgCl₂）,low-dose,middle-dose and high-dose ginkgo total flavonoids groups（1 000 μmol/L HgCl₂+100,200,400 mg/L ginkgo total flavonoids）. After incubation for 24 hours,the cell survival was detected by tetramethylazolyl blue（MTT） method,the level of apoptosis was detected by flow cytometry,the level of autophagy was detected by microtubule-associated protein 1 light chain 3B-Ⅱ（LC3B-Ⅱ） antibody staining,the levels of glutathione,r-glutamyl cysteingl+glycine（GSH） and malondialdehyde（MDA） were measured,and levels of SIRT1 and NLRP3 were detected by real-time fluorescent reverse transcription（RT-PCR） and Western blotting. Results Compared with the control group,the survival rate of cells in the mercury poisoning model group[（93.63±2.51）% vs（48.63±3.25）%],the positive rate of cell LC3B-Ⅱ[（58.25±12.15）% vs（10.19±3.20）%],GSH content[（115.82±15.91）U/mg vs（52.74±6.40）U/mg],SIRT1 mRNA protein expression（2.93±0.80 vs 0.48±0.14,0.30±0.05 vs 0.09±0.03） decreased,and the difference were statistically significant（all P<0.05）. Compared with the mercury poisoning model group,the cell survival rate after ginkgo total flavonoids treatment[（48.63±3.25）% vs（59.28±3.18）%,（74.21±3.63）%,（82.29±3.59）%],the positive rate of cell LC3B-Ⅱ[（10.19±3.20）% vs（15.20±5.33）%,（22.02±8.65）%,（36.22±11.35）%],GSH content[（52.74±6.40）U/mg vs（67.36±9.71）U/mg,（84.52±7.46）U/mg,（92.48±5.29）U/mg],SIRT1 mRNA protein expression（0.48±0.14,0.78±0.23,1.52±0.30 vs 2.20±0.72, 0.09±0.03 vs 0.23±0.06,0.19±0.08,0.14±0.06） increased,and the difference were statistically significant（all P<0.05）. Compared with the control group,the apoptosis rate of mercury poisoning model group[（0.16±0.08）% vs（1.14±0.19）%],MDA content[（5.42±0.68）nmol/mg vs（18.60±3.21）nmol/mg],NLRP3 mRNA protein expression levels（0.35±0.09 vs 3.28±1.02,0.24±0.04 vs 0.73±0.12） significantly increased,and the difference were statistically significant（all P<0.05）. Compared with the mercury poisoning model group,the cell apoptosis rate after ginkgo total flavonoids treatment[（1.14±0.19）% vs（0.83±0.12）%,（0.32±0.07）%,（0.21±0.07）%],MDA content[（18.60±3.21）nmol/mg vs（15.79±4.26）nmol/mg,（10.21±2.47）nmol/mg,（8.16±1.05）nmol/mg],NLRP3 mRNA protein expression levels（3.28±1.02 vs 2.45±0.86,1.53±0.46,0.83±0.32,0.73±0.12 vs 0.60±0.09,0.41±0.06,0.37±0.07） decreased,with the dose-response relationship,and the differences were statistically significant（all P<0.05）. Conclusions The total flavonoids of Ginkgo biloba have a protective effect on the cytotoxicity of human embryonic kidney caused by mercury poisoning,can reduce cell apoptosis and oxidative damage,and enhance cell autophagy. The mechanism may be related to total flavonoids of Ginkgo biloba increasing the expression of SIRT1,reducing the expression of NLRP3,and then inhibiting SIRT1/NLRP3 pathway.

Key words: Mercury poisoning, Total flavonoids of Ginkgo biloba, SIRT1, NLRP3, Human embryonic kidney cells

CLC Number:

R114

ZHANG Jian-xin, HE Qing, ZHANG Jian-bin, LIU Jie. Interventional effect of total flavonoids of Ginkgo biloba on human embryonic kidney cytotoxicity induced by mercury poisoning and its effect on SIRT1/NLRP3 pathway. [J]OCCUPATION AND HEALTH, 2023, 39(9): 1158-1164.

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