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Experimental Study on the Effects of Naofucong on Lipopolysaccharide-activated Synthesis of Cyclooxygenase-2 and Secretion of Prostaglandin E2 by Microglia

Author: CuiYaZhong
Tutor: ZhangMengRen
School: Beijing Union Medical College
Course: Traditional Chinese Medicine
Keywords: Lipopolysaccharide Microglia COX-2 PGE2 Naofucong
CLC: R285.5
Type: Master's thesis
Year: 2011
Downloads: 67
Quote: 0
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Abstract


[Objective]To observe the effects of Naofucong serum on morphology of the mouse microglia BV-2, expression of COX-2 protein in BV-2, and the level of PGE2 secreted in the supernatant of the BV-2 culture after lipopolysaccharide activation.[Methods]Mouse microglia BV-2 cells incubated in vitro were divided into three groups:the normal control group, the LPS group (lμg/ml final concentration for activation) and the high-, moderate-and low-dose of Naofucong (NFC) serum groups. After 24h incubation, the morphology of microglia was observed with inverted phase contrast microscope, the expression of the COX-2 protein was detected by Western blot, and the level of PGE2 protein in the supernatant of BV-2 culture was determined by enzyme-linked immunosorbent assay.[Results]1. morphology of microglia BV-2 cellsAs observed in inverted phase contrast microscopy, in the normal control group, about 70-80% of microglia BV-2 cells were well-adherent and in shape of spindle or triangle,20-30% of the cells were suspended; while after treatment with lμg/ml LPS for 24h, about 10-20% of microglia BV-2 cells were adherent, about 80-90% of the cells were suspended, a larger amount of the cells were aggregated into clumps and some of them became round-shaped. In the high-, moderate-and low-dose of NFC serum groups, the amount of adherent cells increased, indicating a certain restoring effect on the LPS-induced morphological changes.2. expression of COX-2 protein in BV-2As shown by Western blot assay, the level of COX-2 protein in BV-2 in the LPS group increased significantly compared with that of the control group (P<0.01). And the levels of COX-2 protein in BV-2 in all the NFC serum groups decreased obviously compared with that of the LPS group (P<0.01).3. Level of PGE2 protein in the supernatant of BV-2 culture As determined by ELISA, the level of PGE2 of BV-2 in the LPS group increased significantly compared with that of the normal control group (P<0.01). And the levels of all the NFC serum groups decreased obviously compared with the LPS group (P< 0.01).[Conclusions]The NFC serum can down regulate the expression of COX-2 protein in BV-2 activated by LPS significantly and can reduce the secretion of PGE2 in the supernatant of BV-2 culture.[Innovation]With the model of the mouse microglia BV-2 cells incubated in vitro, the effects of a traditional Chinese complex preparation serum on the expression of COX-2 protein in BV-2 activated by LPS and the level of PGE2 secreted in the supernatant of BV-2 culture were observed in the cellular level, providing an experimental support for the clinical prophylaxis and treatment of the degenerative disease by Naofucong.

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