Regulatory effect and significance of hydrogen sulfide on low-density lipoprotein receptor / 中华实用儿科临床杂志

ABSTRACT

Objective To observe the regulatory effect and significance of hydrogen sulfide (H2 S)on low -density lipoprotein receptor (LDLR).Methods Mouse primary hepatocytes were divided into control group,low -den-sity lipoprotein (LDL)group and LDL +sodium hydrosulfide (NaHS,the donor of H2 S)group.The cells in LDL group were treated with LDL (50 mg/L)and the cells in LDL +NaHS group were pretreated with NaHS (100 μmol/L)for 0.5 hours and then treated with LDL (50 mg/L).Real -time PCR and Western blot were used to detect the expres-sions of LDLR mRNA and protein,respectively.Mouse primary hepatocytes were divided into control group,1,1′-dioctadecyl -3,3,3′,3′-tetramethyl -indocarbocyanine perchlorate low -density lipoprotein (DiI -LDL)group and DiI -LDL +NaHS group.The cells in DiI -LDL group were incubated with DiI -LDL (10 mg/L)for 3 hours.The cells in DiI -LDL +NaHS group were pretreated with NaHS (100 μmol/L)for 1 hour before DiI -LDL (10 mg/L) was added.Confocal method was used to measure the uptake of DiI -LDL by mouse primary hepatocytes,and fluores-cent quantitative method was performed to detect the content of DiI -LDL in the supernatant of mouse primary hepato-cytes.Results The levels of LDLR mRNA and protein in the mouse primary hepatocytes were significantly downregu-lated compared with those in the control group (t =5.733,P <0.01;t =2.527,P <0.05);after NaHS was adminis-tered,LDLR mRNA and protein level in the mouse primary hepatocytes were significantly upregulated (t =-7.639, P <0.01;t =2.388,P <0.05).In the mouse primary hepatocytes,compared with that in the control group,the uptake of DiI -LDL by cells in DiI -LDL group was increased;the uptake of DiI -LDL by mouse primary hepatocytes in DiI -LDL +NaHS group was significantly increased in comparison with that in the DiI -LDL group.Compared with that in the control group,the DiI -LDL content of culture supernatant in the DiI -LDL group was significantly increased (t =-39.156,P <0.01);after treatment with H2 S donor,the content of DiI -LDL in the culture supernatant was sig-nificantly decreased in comparison with that in the mouse primary hepatocytes without H2 S donor treatment (t =17.202,P <0.01).Conclusion H2 S upregulated the expression of LDLR protein in the mouse primary hepatocytes.