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Objective@#To establish a new model of hepatic steatosis cells by optimizing the original ethanol or high fat, the present study proposed an in vitro hepatocyte steatosis model for the study of fatty liver.@*Methods@#Oil red O staining was used to observe the effects of fetal bovine serum, oleic acid and ethanol on lipid accumulation in human liver cell line L02 in a concentration- and time-dependent manner. RT-PCR was used to detect the mRNA expression levels of PPAR-γ and AP-2, and the suitable conditions for the establishment of hepatocyte steatosis model were screened out. A t-test was used for comparison between the two groups, and one-way Analysis of Variance (ANOVA) was used in more than three groups.@*Results@#Oil red O staining showed the number of reddish-orange lipid droplets in L02 cells gradually increased with the increase of fetal bovine serum, oleic acid and ethanol in a concentration - and time-dependent manner. Compared with 0.00% oleic acid and 2% ethanol, the count value of red particle was 100.00% ± 17.63% at the beginning and after 24 h, 0.003% oleic acid and 2% ethanol jointly acted in L02 cells. After incubation for 48 hours with 2% ethanol and serum-free DMEM medium, the accumulation of lipid droplets was the highest with a count value of 802.38%+71.06%(t = 42.36, P < 0.001). RT-PCR analysis showed the lipid accumulation induced by this method was positively correlated with the mRNA expression of PPAR-γ and AP-2.@*Conclusion@#L02 cells were successfully exposed to high fat and ethanol, and the hepatocyte steatosis model was established and optimized, suggesting that the occurrence of hepatic cell steatosis was related to the up-regulation of PPAR-γ and AP-2.
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AIM: To investigate the effect of injection stauntoniae ( IS) on inflammatory pain responses in mice.METHODS:The carrageenan test was used to determine the anti-inflammatory and analgesic effects of IS .Except for control group, the mice in other groups received an injection of λ-carrageenan solution (1%, 50 μL) into the plantar region of the left hind paws , followed by a subcutaneous injection of IS at doses of 12.5%, 50%and 100%or equal vol-ume of 0.9%NaCl.Both paw edema and hyperalgesia to thermal stimulation were measured 4 h, 12 h, 24 h and 48 h after the injection of λ-carrageenan solution.The lumbar-5 (L5) dorsal root ganglions (DRGs) of the mice were taken to inves-tigate the cyclooxygenase 2 ( COX-2) expression by immunohistochemical staining .RESULTS:Subcutaneous injection of IS potently inhibited paw edema and hyperalgesia induced by λ-carrageenan in the mice accompanied with the inhibition of COX-2 protein expression in L 5 DRGs.CONCLUSION:IS exerts the anti-inflammatory and analgesic effects on the in-flammatory responses by inhibiting the protein expression of COX-2 in DRGs.