Modulation of GdCl3 and Angelica sinensis polysaccharides on differentially expressed genes in liver of hepatic immunological injury mice by cDNA microarray.
OBJECTIVE
To study the modulating effect of GdCl(3) and Angelica Sinensis polysaccharides (ASP) on differentially expressed genes in liver of hepatic immunological mice by cDNA microarray.


METHODS
Hepatic immunological injury was induced by lipopolysaccharide (LPS ip, 0.2 mg/kg(-1)) in bacillus calmetteguerin (BCG ip, 1 mg/kg(-1)) primed mice; A single dose of 20 mg/kg(-1) GdCl(3) was simultaneously pretreated and 30 mg/kg(-1) ASP (ig, qdX7 d) was administrated when the BCG+LPS was primed. The mice were sacrificed at the end of the 7(th) day after ip LPS for 6 h and the liver was removed quickly. The PCR products of 512 genes were spotted onto a chemical material-coated glass plate in array. The DNAs were fixed to the glass plate after series of treatments. The total RNAs were isolated from the liver tissue, and were purified to mRNAs by Oligotex. Both mRNAs from the normal liver tissue and the liver tissue from the mice with hepatic immunological injury or that pretreated with GdCl(3) or ASP were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for fluorescent signals and showed differences between the two tissues.


RESULTS
Among the 512 target genes, 18 differed in liver tissue of hepatic immunological injury mice, and 6 differed in those pretreated by ASP, 7 differed in those pretreated by GdCl(3).


CONCLUSIONS
cDNA microarray technique is effective in screening the differentially expressed genes between two different kinds of tissue. Further analysis of those obtained genes will be helpful to understand the molecular mechanism of hepatic immunological injury and to study the intervention of drug. Both ASP and GdCl(3) can decrease the number of the differentially expressed genes in liver tissue of mice with hepatic immunological injury.
Ding
Hong
H
Medical College, Shantou University, Guangdong Province, China. dinghong2000@263.net
Shi
Gang-Gang
GG
Yu
Xin
X
Yu
Jie-Ping
JP
Huang
Jie-An
JA
D029971:Angelica sinensis; D000818:Animals; D005682:Gadolinium / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D008070:Lipopolysaccharides / Q000633:toxicity; D008099:Liver / Q000187:drug effects* / Q000293:injuries / Q000378:metabolism*; D051379:Mice; D009163:Mycobacterium bovis / Q000276:immunology; D020411:Oligonucleotide Array Sequence Analysis; D011134:Polysaccharides / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D008070:Lipopolysaccharides; D011134:Polysaccharides; D012333:RNA, Messenger; D005682:Gadolinium; C038958:gadolinium chloride