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Toll-like-receptor-4 Signal Pathway in Vascular Damage in Kawasaki Disease

Author: FanXiaoWei
Tutor: HuXiuFen
School: Huazhong University of Science and Technology
Course: Pediatrics
Keywords: TLR4 Kawasaki disease Gamma globulin
CLC: R725.4
Type: Master's thesis
Year: 2010
Downloads: 20
Quote: 0
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Abstract


Objective To observe the Kawasaki disease (KD) and acute phase serum gamma globulin on monocyte / macrophages TLR4 (Toll-like-receptor 4) signal transduction pathway molecule gene expression to explore TLR4 signaling pathway in vascular injury KD Role and gamma globulin therapy KD possible mechanism. Method 1. Experimental groups: monocytes / macrophages (THP1) grouped according to the following design: ⑴ normal serum group: RPMI1640 medium 10% of healthy children serum; ⑵ K D serum group: ① KD-CAL-serum group: RPMI1640 medium 10 % KD-CAL-acute phase serum; ② KD-CAL serogroup: RPMI1640 medium 10% KD-CAL acute phase serum; ⑶ gamma globulin group: ① KD-CAL-gamma globulin group: RPMI1640 medium 10% KD-CAL - Acute phase serum gamma globulin 20mg/ml; ② KD-CAL gamma globulin group: RPMI1640 medium 10% KD-CAL acute phase serum gamma globulin 20mg/ml. 2. THP-1 monocyte-derived macrophages culture: human monocyte-macrophage cell line (THP-1) were grown by the typical center of Wuhan University (CCTCC) provided, containing 10% fetal bovine serum, 0.5% two mercaptoethanol in RPMI1640 culture medium, 37. C, 5% CO2 under static culture conditions. Logarithmic growth phase cells, adjusting a density of 1 × 109 / L were seeded in 6-well plates with 160nmol / L PMA THP-1 cells were incubated for 24 hours, 80% of the cells seen by the suspension becomes adherent state, and cell shape fusiform from round, oval or irregular in shape, has prompted monocytes into macrophages. Into the above culture group (n = 6 wells each) in each cell culture plate medium was added to the above each set of conditions, cultured for 24h, the solution with 0.25% trypsin digested cell equipment seized. 3 using reverse transcription polymerase chain reaction (RT-PCR) method on monocyte / macrophages TLR4, MyD88, TRAF6, CD14 mRNA was amplified agarose gel electrophoresis, gel imaging analysis system electrophoretic bands measured density values ??to G3PDH as an internal reference, calculating the relative content of the target gene mRNA, and results in the target gene and G3PDH said band density ratio. Results 1. KD serum group with normal serum group THP-1 cells TLR4 signaling pathway molecules comparison of gene expression: KD serogroup THP-1 cells TLR4, MyD88, TRAF6 mRNA expression was significantly higher than the normal serum group [(1.27 0.17)% group, the difference was not statistically significant (P gt; 0.05). gamma globulin serum group compared with KD, TLR4, MyD88, TRAF6 mRNA expression was decreased to varying degrees. 2. KD-CAL-serum and serum KD-CAL group, THP-1 cells TLR4, MyD88, TRAF6, CD14 mRNA expression showed no significant difference (P gt; 0.05). Conclusions TLR4 pathway in vascular inflammatory injury Kawasaki disease plays an important role in gamma globulin by blocking TLR4 pathway may play a role in treatment of Kawasaki disease.

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CLC: > Medicine, health > Pediatrics > Children within the science > Pediatric cardiac and vascular disease
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