[Effects of soothing liver and invigorating spleen recipes on LPS-induced hepatocytes injury of rats and TLR4/p38MAPK signal pathway].
OBJECTIVE
To explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.


METHODS
The hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.


RESULTS
The rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.


CONCLUSIONS
The soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.
Xiang-Wen
XW
Gong
Qin-He
QH
Yang
Hai-Zhen
HZ
Yan
Yu-Pei
YP
Zhang
Yin-Ji
YJ
Liang
Yi-Zhen
YZ
Liu
Zhang Jin-Wen
Chun-Mei
CM
Lin
Yuan-Yuan
YY
Li
D000818:Animals; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage*; D005260:Female; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008070:Lipopolysaccharides / Q000009:adverse effects; D008099:Liver / Q000187:drug effects* / Q000293:injuries / Q000378:metabolism; D008297:Male; D065626:Non-alcoholic Fatty Liver Disease / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D013154:Spleen / Q000187:drug effects* / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*
D004365:Drugs, Chinese Herbal; D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4; D048051:p38 Mitogen-Activated Protein Kinases