[DOI] /j.issn China

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1 Med J Chin PL, Vol., No., pril, 8 89 [ ]() PC g/ml PC6 h 6(IL-6) (TNF- ) Toll(TLR) 88(MyD88) p-p65( 6h) () (g/ml ) ( mol/l ) ( g/ml mol/l ) IL-6 TNF-6 h (P<.5) 6h PCTLR/MyD88/NF- B 6hPC IL-6 TNF- TLR MyD88 p-p65 (P<.5) PC [] PCToll [ ] R6 R76.6 [ ] [ ] 577-7(8)-89-5 [DOI].855/j.issn Protective role and mechanism of dexmedetomidine on the -stimulated PC cells ZHOU Jun-jie, XIO Wei, HE Xuan, PENG Na, TONG Hua-sheng *, SU Lei Department of Intensive Care Unit, Heyuan People's Hospital ffiliated to Jinan University, Heyuan, Guangdong 57, China Department of Intensive Care Unit, Guangzhou General Hospital of Guangzhou Military rea Command, Guangzhou 5, China * Corresponding author, fimmuths@6.com This work was supported by the Doctoral Start-up Projects of Natural Science Foundation of Guangdong Province (688) [bstract] Objective To explore the effect of dexmedetomidine on the lipopolysaccharide () stimulated PC cells and its potential mechanism. Methods PC cells were treated by with a concentration of g/ml. The cell viability, the concentrations of interleukin-6 (IL-6) and tumor necrosis factor (TNF- ) in the cell culture supernatant were measured after -, 6-, or -h treatment. The expressions of toll-like receptor (TLR), myeloid differentiation factor 88 (MyD88) and phosphorylated p65 (p-p65) were measured. In the second part, PC cells were cultured under four different treatments, that is, normal culture media in first group, g/ml in second group, mol/l dexmedetomidine in third group, g/ml and mol/l dexmedetomidine in fourth group. The indexes mentioned above were measured 6 hours after and treatments. Results The cell viability was decreased after treatment, and the concentrations of IL-6 and TNF- were increased significantly. Compared with control group, the concentrations in -, 6-, -h groups showed statistically significant differences (P<.5), especially after 6 hours. The TLR/MyD88/NF- B pathway was activated after stimuli and reached the peak value. Compared with treatment group, PC cell apoptosis rate, the concentrations of IL-6 and TNF- and the expressions of TLR, MyD88 and p-p65 were decreased. The differences between group and group was statistically significant (P<.5). Conclusion Dexmedetomidine has a protective effect on stimulated PC cells via the inhibition of inflammatory response. [Key words] lipopolysaccharide; dexmedetomidine; PC cells; Toll-like receptor [ ] (688) [ ] MODS [ ] 57 () 5 () [ ] fimmuths@6.com

2 9 8 [] [] [] [] (lipopolysaccharide ) [5] Tol l (Tolllike receptors TLR) TLR (pathogen-associated molecular patterns PMPs) TLR88 (myeloid differentiation factor 88 MyD88) NF- B [6] (dexmedetomidine ) [7] ICU [8-9] [] PC. MTT Hoechst MPDMEM GibcoTNF- IL-6 ELIS ebiosiencetlr MyD88 Cell Signaling Technology.PC 7 5% CO % %- DMEM h 6 h [] g/ml 6 h g/ml [] mol/l g/ml mol/l.96 MTT 96.5mg/ml MTT 5 l 7 h MTT DMSO 5 l min 9nm(OD) = OD /OD %. Hoechst % 5min PBS g/ml Hoechst 7min 6nm nm.5 TNF- IL-6 5nm OD TNF- IL-6.6TLR MyD88 p-65 p-p65 Western blotting BCSDS-PGE TLR(:) MyD88(:) p-65(:) p-p65(:) (:5) Image J.7SPSS. x±s LSD-tP<.5. PCIL-6 TNF- g/ml PC 6h h 6h (P<.5) IL-6 TNF- 6 h (P<.5) 6h ( ). PC TLR MyD88 p-p65 Western blotting PC TLR MyD88 p-p65 ( ) 6h IL-6 TNF- 6h

3 Med J Chin PL, Vol., No., pril, B C PC () IL-6(B) TNF- (C) Fig. Effects of on PC cell viability (), concentrations of IL-6 (B) and TNF- (C) at different time points Cell viability (%) TLR GPDH IL-6 (pg/ml) P<.5 compared with control group MyD88 GPDH B p65 C PC TLR() MyD88(B) p-p65(c) Fig. Effects of on the expressions of TLR (), MyD88 (B) and p-p65 (C) in PC cells at different time points TNF- (pg/ml) p-p65. PC g/ml mol/l PC (P<.5) Hoechst ( B) PC Cell viability (%) 5 5 B PC 6h () (B, Hoechst staining, ) Fig. Effects of dexmedetomidine on the cell viability () and apoptosis rate (B, Hoechst staining, ) of -treated PC cells P<.5 compared with control group; P<.5 compared with group. PC IL-6 TNF- IL pg/ml 58.7.pg/ml (P>.5)6h pg/ml (P<.5) 6h IL pg/ml ( P <.5) TNF- IL-6 (pg/ml) pg/ml pg/ml ( P >.5)6h pg/ml(P<.5) 6h TNF pg/ml (P<.5 ) PC.5 PC TLR MyD88 p-p65 Western blotting PC 6h IL-6() TNF- (B) Fig. Effects of dexmedetomidine on the concentrations of IL-6 () and TNF- (B) in the supernatants of -treated PC cells P<.5 compared with control group; P<.5 compared with group TNF- (pg/ml) 6 B

4 9 8 PCTLR MyD88 p-p65 Image J TLR/GPDH TLR GPDH 5 MyD88/GPDH MyD88 GPDH (P<.5 5) B C 5 PC 6hTLR() MyD88(B) p-p65(c) Fig.5 Effects of dexmedetomidine on the expressions of TLR (), MyD88 (B) and p-p65 (C) in -treated PC cells p-p65/p65 p-p65 p65 P<.5 compared with control group; P<.5 compared with group 5 [] ICU - [-7] 8d [8] [9] TNF- IL-6 IL-8 [-] Ning [] TNF- IL-6 PC TLR TLR/MyD88/NF- B TNF- IL-6 Toll TLR Toll [-] TLR (PMPs) TLR MyD88 IL-(interleukin- receptor associated kinase IRK) 6(TNF receptor associated factor 6 TRF6) NF- B TLR/MyD88/NF- B TNF- IL-6 Liu [] TLR/MyD88/NF- B [5-6] TNF- IL- [7] TLR/MyD88/NF- B TNF- IL-6 c-jun TNF- IL-6 [8] PC TNF- IL-6

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