• Archives of Pharmacal Research
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• 제26권7호
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• pp.554-558
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• 2003

Platelet activating factor acetylhydrolase (PAF-AH) activity has been identified in cerebrospinal fluid (CSF) samples taken from children with meningitis. We reported that PAF-AH activity is significantly increased, by about 3 fold, in patients with meningitis compared to control subjects. Because of limited knowledge about this enzyme in CSF, we examined the biochemical properties of CSF PAF-AH. PAF-AH of CSF was calcium independent, showed a broad pH spectrum and was relatively heat stable. In addition, this enzyme activity was strongly inhibited by phenylmethanesulfonyl fluoride (PMSF), partially inhibited by p-bromophenacylbromide (p-BPB), uninhibited by iodoacetamide, and moderately stimulated by dithiothreitol (DTT). PAF-AH of CSF did not degrade phospholipid with a long chain fatty acyl group at sn-2 position. This enzyme hydrolyzed PAF and oxidatively modified phosphatidylcholine. Furthermore, we identified a monomeric polypeptide with a molecular weight of approximately 45 kDa by Western blot using human plasma PAF-AH antibody. These results suggested that plasma type PAF-AH activity exist in CSF taken from children with meningitis.

• Archives of Pharmacal Research
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• 제20권3호
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• pp.218-224
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• 1997

Platelet-activating factor (PAF) acetylhydrolase, which removes the acetyl moiety at the sn-2 position, has been found in human amniotic fluid. We purified this enzyme by ammonium sulfate precipitation, and sequential use of DEAE-Sepharose CL-6B, hydroxyapatite, chelating-Sepharose, and Mono Q column chromatographies. This enzyme exhibited broad pH optima and was unaffected by EDTA. Partially purified enzyme had a molecular weight of approximately 34 kDa on SDS-PAGE. In addition, the enzyme activity was inhibited by either diisopropylfluorophosphate(DFP) or p-bromophenacylbromide (p-BPB), suggesting that this enzyme possesses active serine and histidine residues. The enzyme showed similar activity towards PAF and oxidatively modified phosphatidylcholine, but didn't hydrolyze phosphatidylcholine or phosphatidylethanolamine with a long chain fatty acyl group at sn-2 position.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1292-1300
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• 2006

Aeromonas encheleia, a potential human intestinal pathogen, was shown to infect a human intestinal epithelial cell line (Caco-2) in a noninvasive manner. The transcriptional profile of the Caco-2 cells after infection with the bacteria revealed an upregulated expression of genes involved in chloride secretion, including that of phospholipase A2 (PLA2) and platelet-activating factor (PAF) acetylhydrolase (PAFAH2). This was also confirmed by a real-time RT-PCR analysis. As expected from PLA2 induction, PAF was produced when the Caco-2 cells were infected with the bacteria, and PAF was also produced when the cells were treated with a bacterial culture supernatant including bacterial extracellular proteins, yet lacking lipopolysaccharides. Bacterial aerolysin was shown to induce the production of PAF.

• Journal of Yeungnam Medical Science
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• 제25권1호
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• pp.41-49
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• 2008

한탄바이러스가 혈소판활성인자 활성에 영향을 미치는지 알아보기 위하여 간접적으로 혈소판활성인자 수용체의 발현과 분해효소의 활성을 측정하였다. 혈관내피세포에서 혈소판활성인자 수용체의 유전자를 역전사 중합효소연쇄반응으로, 단백질은 western blot으로 측정하였다. 또한 세포표면에 발현된 혈소판활성인자 수용체의 양은 FACS로 분석하였다. 한탄바이러스에 감염된 혈관내피세포에서 혈소판활성인자 수용체의 유전자, 단백질, 세포 표면의 발현 모두 바이러스에 감염되지 않은 대조 세포보다 감염 후 2, 3일째 증가 하였다. 혈액 내 혈소판활성인자 분해효소의 활성을 비교한 결과 신증후출혈열 환자에서 정상인에 비하여 2.5배 낮았다. 그리고 신증후출혈열 환자가 회복됨에 따라 혈소판활성인자의 활성이 다시 정상 수준으로 회복되는 것으로 나타났다. 따라서 한탄바이러스에 의해 증가된 혈소판활성인자 수용체의 발현이 혈소판활성인자와 혈관내피세포와 반응성을 증가시키고, 신증후출혈열 환자 혈액에서 감소된 혈소판활성인자 분해효소가 혈소판활성인자의 분해를 지연 시켜 작용시간을 증가 시킴으로써 과다한 혈소판활성인자의 생물학적 작용이 신증후출혈열의 병리현상을 초래할 것으로 사료된다.