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Objective To investigate the effect of fatty acid binding protein 4(FABP4)DNA methylation on abnormal lipid metabolism in placental trophoblastic dyslipidemia. Methods Human placental trophoblast cell line(HTR-8)was treated with L-NAME of 100 μmol/L for 48 h. The lipid content in placental trophoblasts was detected by chemical enzyme-colorimetry. The FABP4 DNA methylation level in placenta trophoblasts was detected by nested-touch down methylation specific PCR (NT-MSP). the mRNA and protein expression of DNMT1 and FABP4 were detected by qRT-PCR and Western Blot,respectively,in trophoblast cells. Results The lipid content in trophoblasts significantly increased as compared with the control(P < 0.05). Expression of FABP4 mRNA and protein increased(P < 0.05),while FABP4 methylation level and expression of DNMT1 significantly decreased (P<0.05)after treatment with L-NAME. Conclusions FABP4 DNA methylation is involved in the regulation of lipid metabolism in placental trophoblastic cells of hypertensive disorder complicating pregnancy.
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Aim To explore the effect of miRNA-143 ( miR-1 4 3 ) on homocysteine ( Hcy ) induced-vascular smooth muscle cells ( VSMCs ) proliferation and the mechanism .Methods VSMCs were cultured and in-cubated with Hcy by using primary cultured method . Then, cells were treated with different concentrations of Hcy and folate .VSMCs proliferation was determined with MTT assay , miR-143 was measured by qRT-PCR, and methylation of miR-143 was determined with meth-ylated PCR.Results After cells were treated with dif-ferent concentrations of Hcy , the proliferation of VSMCs was significantly increased , mRNA expression of miR-143 was decreased and methylation of miR-143 was increased .The proliferation of VSMCs was signifi-cantly decreased when transfected VSMCs with miR-143 precursor , and cell proliferation was increased by using miR-143 inhibitor transfection .Conclusion Hy-pomethylation of miR-143 may inhibit VSMCs prolifera-tion.
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Objective To investigate the function of CFTR in ApoE-/- mice with HHcy-induced hepato-cellular injury. Methods Thirty six 5-week old ApoE-/- mice were divided into three groups , including the ApoE-/- group, the HHcy group and the intervention group, (n = 12). Twelve normal C57BL/6J mice were fed with regular mouse diet as the normal control (SPF grade). HL-7702 human liver cells were intervened by Hcy (100 μmol/L) and 100 μmol/L Hcy + folic acid (100 μmol/L Hcy + F). The changes of Hcy, ALT and AST in the serum and the expression of CFTR mRNA and protein in liver and liver cells were detected. The concen-trations of ALT and AST in the liver cell intervened by VX-770 agonist and CFTR(inh)-172 inhibitor were mea-sured by ELISA. Results Compared with the control group , the levels of Hcy , ALT and AST were higher and the levels of CFTR mRNA and protein were lower in the Meth group (P < 0. 05 ) , while the reverse result in the Meth + F group (P < 0.05). Compared with the control group, the levels of CFTR mRNA and protein were de-creased and the levels of ALT and AST were increased in the 100 μmol/L Hcy group (P < 0.05). Compared with the 100 μmol/L Hcy group , the levels of CFTR mRNA and protein were increased and the levels of ALT and AST were decreased in the 100 μmol/L Hcy + F group (P < 0.05). Stimulated with VX-770 can reduce the concentrations of ALT and AST and the vice versa in the CFTR (inh)-17 group the concentration was increased in liver cells. Conclusion CFTR plays an important role in the regulation of hepatocellular injury by HHcy.
ABSTRACT
Objective To study the effect of homocysteine(Hcy)on the formation of atherosclerotic and acceleration of ApoE-/-mice liver lipid metabolism disorder .Methods 12 normal 5 weeks old C57BL/6J mice served as control group ,and 36 5 weeks old C57BL/6J A poE-/- mice were randomly divided into 3 groups(n=12 for each group) ,the model control group ,the hyperhomocys-teinemia(HHcy)group and the intervention group(intervened by folate and vitamin B12 ) .18 weeks later ,the blood of the mice was gotten using a Unilateral enucleation method ,and the Serum Hcy and lipid changes were detected by Biochemical analyzer .And the changes of plaque size were measured by HE staining .The liver tissues of the 4 groups mice were taken and the changes in hepato-cyte lipid were detected by oil red O staining ,and the hepatic lipid levels were measured by enzymatic determination(by Semi-quan-titative image analysis) .Results The results showed that ,when compared with the control group ,the serum Hcy ,LDL ,TG and CHOL levels of the HHcy group significantly increased by 2 .3 ,2 .8 ,5 .0 ,10 .7 fold(P<0 .01)and the content of HDL decreased by 64% (P<0 .01) ,and the result showed that ,conpared with the HHcy group ,the seram Hcy ,LDL and CHOL levels of the interven-tion group were significantly decreased by 43% ,34% ,21% (P<0 .05) .Atherosclerotic fatty plaque could be seen in the hyperlipi-demic ,model and intervention group .Meanwhile ,there was a large number of scattered fat in A poE-/-mice liver by oil red O staining in the HHcy group ,and the CHOL and TG levels were 2 .2 fold and 2 .8 fold higher in the HHcy than that in the normal control group respectively(P<0 .01) .And compared with the HHcy group ,the serum CHOL and TG levels of the intervention group sig-nificantly decreased by 34% ,33% (P<0 .01) .Conclusion It is found that Hcy can induce the formation of As and accelerate liver lipid metabolism disorder .