Cholesterol metabolism and expression of its relevant genes in cultured steatotic hepatocytes.
OBJECTIVE
To investigate the cholesterol metabolism and mRNA expression of the relevant genes in cholesterol synthesis of the cultured steatotic hepatocytes model.


METHODS
A steatotic model of hepatocytes was constructed by adding palmitic acid to the growing L-02 cells. These cells were collected at day 3 and 6, respectively. Cells with the culture solution without palmitic acid added served as the control. The contents of intracellular triglyceride (TG) and total cholesterol (TC) were detected by the analysis kit. The expression of sterol-regulatory element binding protein-2 (SREBP-2) and its target gene hydroxymethylglutaryl CoA reductase (HMGCR) and low-density lipoprotein receptor (LDLR) were measured by RT-PCR.


RESULTS
Hepatocyte steatosis was observed at day 3 and became more intense at day 6. The contents of intracellular TG and TC were increased and the expression of the SREBP-2, HMGCR and LDLR mRNA were upregulated in a time-dependent manner in the model group. Compared with the control group, the content of intracellular TG was higher at both day 3 and 6 (P < 0.05), while the content of intracellular TC was significantly increased only at day 6; The expression of HMGCR and LDLR mRNA was upregulated in steatotic hepatocytes at both day 3 and 6 (P < 0.05), whereas the SREBP-2 mRNA was increased only at day 6 (P < 0.05).


CONCLUSIONS
Cholesterol accumulation is probably due to the upregulated expression of the relevant genes in the cholesterol synthesis of the steatotic hepatocytes.
Wang
Xiao Hong
XH
Department of Gastroenterology, The Second Affiliated Hospital of Xuzhou Medical College (The General Hospital of Xuzhou Coal Mineral Group), Xuzhou, Jiangsu, China.
Tian
Yan
Y
Guo
Zhao Jun
ZJ
Fan
Zhu Ping
ZP
Qiu
De Kai
DK
Zeng
Min De
MD
D045744:Cell Line, Tumor; D002784:Cholesterol / Q000096:biosynthesis / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005234:Fatty Liver / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D015870:Gene Expression / Q000502:physiology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics*; D050356:Lipid Metabolism / Q000235:genetics; D019308:Palmitic Acid / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011973:Receptors, LDL / Q000235:genetics*; D051782:Sterol Regulatory Element Binding Protein 2 / Q000235:genetics*
D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D011973:Receptors, LDL; C497390:SREBF2 protein, human; D051782:Sterol Regulatory Element Binding Protein 2; D019308:Palmitic Acid; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases