Effect of JIANPI HUOXUE decoction on inflammatory cytokine secretion pathway in rat liver with lipopolysaccharide challenge.
OBJECTIVE
To evaluate the effect of Chinese traditional medicinal prescription, JIANPI HUOXUE decoction (JHD) on cytokine secretion pathway in rat liver induced by lipopolysaccharide (LPS).


METHODS
Twenty-four male SD rats were divided into normal group (n = 4), model group (n = 10) and JHD group (n = 10) randomly. Rats in model group and JHD group were administrated with normal saline or JHD via gastrogavage respectively twice a day for 3 d. One hour after the last administration, rats were injected with LPS via tail vein, 50 mug/kg. Simultaneously, rats in normal group were injected with equivalent normal saline. After LPS stimulation for 1.5 h, serum and liver tissue were collected. Pathological change of liver tissues was observed through hematoxylin-eosin (H.E.) staining. Tumor necrosis factor alpha (TNF-alpha) in serum were assayed by enzyme linked immunosorbent assay (ELISA). The protein expression of TNF-alpha, phosphorylated inhibit-kappaB (p-IkappaB) and CD68 in liver were assayed by Western blot. The distribution of CD68 protein in liver was observed through immunohistochemical staining. The mRNA expression of TNF-alpha, interleukin-6 (IL-6), CD14, toll-like receptor 2 (TLR2) and TLR4 in liver were assayed by real-time RT-PCR.


RESULTS
Predominant microvesicular change, hepatocyte tumefaction and cytoplasm dilution were observed in liver tissues after LPS administration as well as obvious CD68 positive staining in hepatic sinusoidal. After LPS stimulation, serum TNF-alpha (31.35 +/- 6.06 vs 12225.40 +/- 9007.03, P < 0.05), protein expression of CD68 (1.13 +/- 0.49 vs 3.36 +/- 1.69, P < 0.05), p-IkappaB (0.01 +/- 0.01 vs 2.07 +/- 0.83, P < 0.01) and TNF-alpha (0.27 +/- 0.13 vs 1.29 +/- 0.37, P < 0.01) in liver and mRNA expression of TNF-alpha (1.96 +/- 2.23 vs 21.45 +/- 6.00, P < 0.01), IL-6 (4.80 +/- 6.42 vs 193.50 +/- 36.36, P < 0.01) and TLR2 (1.44 +/- 0.62 vs 4.16 +/- 0.08, P < 0.01) in liver were also increased significantly. These pathological changes were all improved in JHD group. On the other hand, TLR4 mRNA (1.22 +/- 0.30 vs 0.50 +/- 0.15, P < 0.05) was down-regulated and CD14 mRNA increased but not significantly after LPS stimulation.


CONCLUSIONS
JHD can inhibit cytokine secretion pathway induced by LPS in rat liver, which is probably associated with its regulation on CD68, p-IkappaB and endotoxin receptor TLR2.
Peng
Jing-Hua
JH
Institute of Liver Disease, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, No. 528, Zhangheng Road, Shanghai 201203, China.
Hu
Yi-Yang
YY
Cheng
Yang
Y
Han
Chong
C
Xu
Li-Li
LL
Feng
Qin
Q
Chen
Shao-Dong
SD
Tao
Qing
Q
Li
Hong-Shan
HS
Li
Xue-Mei
XM
D000818:Animals; D016207:Cytokines / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000494:pharmacology*; D005260:Female; D008070:Lipopolysaccharides / Q000276:immunology*; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley
D016207:Cytokines; D004365:Drugs, Chinese Herbal; C528003:Jianpi Huoxue decoction; D008070:Lipopolysaccharides; D012333:RNA, Messenger