肥大细胞对THP-1巨噬细胞源性泡 沫细胞胆固醇分配和流出的影响
赵战芝 杨永宗
南华大学心血管病研究研
肥大细胞(MCs)
一种参与炎症免疫应答反应的炎症细胞。
胞浆内含有大量异染性颗粒（组胺等）。
受刺激时脱颗粒,释放胞浆一系列可溶性介质到细胞外 （compound48/80、oxLDL等可以刺激MCs脱颗粒) 。
研究背景
促动脉粥样硬化(As)形成
MCs加速oxLDL诱导平滑肌细胞泡沫化。
Compound48/80处理的新西兰白兔和apoE-/-鼠主动脉脂纹形成比对照组早，面积大，分布广。
HDL Metabolism and Reverse Cholesterol Transport
胆固醇流出减少
A-I
CE
CE
CE
FC
FC
LCAT
FC
Bile
SR-BI
A-I
ABCA1
Mature HDL
Nascent HDL
MC
?
ABC1 = ATP-binding cassette protein 1; A-I = apolipoprotein A-I; CE = cholesteryl ester; FC = free cholesterol; LCAT = lecithin:cholesterol acyltransferase; SR-BI = scavenger receptor class BI
实验结果
A
B
The effect of compound 48/80 on MCs degranulation
MCs were stained by toluidine blue (×1000)
A，MCs；
B，MCs+C48/80.
MCs histamine release percentage
Formation of THP-1 macrophage-derived foam cells
CE/TC Ratio:68.7%
HPLC analysis of THP-1macrophage -derived foam cells
Oil red O staining of THP-1macrophage -derived foam cells
FC，free cholesterol
CE，cholesterol ester
FC
CE
The effect of MCs on the accumulation of cell cholesterol in THP-1 macrophage-derived foam cells
Values are represented as（ ）mg cholesterol/g protein, ( n=5 ).
a:P<0.05 vs.control,HDL3+MCs group;
b:P<0.05 vs. control group;
c: P<0.05 vs. HDL3 group.
Cholesterol standard
control
HDL3
MCs
HDL3+MCs
C48/80
HDL3+C48/80
The chromatography picture of cholesterol and cholesterol ester in THP-1 macrophage-derived foam cells identified by high performance liquid chromatography.
The effect of MCs on the distribution of cell free and esterified cholesterol in THP-1 macrophage-derived foam cells
control (alone)=69.2%;
with HDL3=47.5%;
with MCs = 27.1%;
with HDL3+ MCs =7.6%;
with compound 48/80=57.5%;
and with compound 48/80+HDL3=19.4%. a:P<0.05 vs. control group;
b: P<0.001 vs. HDL3 group.
n=5.
The effect of MCs on cholesterol efflux from THP-1 macrophage-derived foam cells mediated by HDL3
control=2.89%;
with HDL3=40.98%;
with MCs =2.91% ;
with HDL3+ MCs =26.92%;
with compound 48/80=3.71%;
and with compound 48/80+HDL3=35.23%.
The values are mean±SD (n=5).
** P<0.01 vs. HDL3+MCs group ;
# P<0.05 vs. HDL3 +MCs group ;
* P<0.001 vs. HDL3 group .
GAPDH
(697bp)
ABCA1
(306bp)
Marker control HDL3 MCs HDL3+MCs C48/80 HDL3+C48/80
*
The effect of MCs on ABCA1 mRNA expression in THP-1 macrophage-derived foam cells
After the treatment of MCs, no significant changes were shown
in transcription level of cell ABCA1 compared with control group.
Compared with control group , * P >0.05.
KDa
31
25
19
6
A 1 2
B St 1 2
15% SDS-PAGE[wt/vol]
AGE
Degradation of HDL3 and apoA-I by MCs
1, untreated HDL3; 2, MCs -treated HDL3.
MCs能降解HDL3的apoA-Ⅰ；
MCs能抑制HDL3介导的 THP-1巨噬细胞源性泡沫细胞胆固醇流出，促使细胞内游离胆固醇蓄积。
结论