Limits...
NLRP3 inflammasome: a novel link between lipoproteins and atherosclerosis

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Pattern recognition receptor-mediated signaling pathways have recently been elucidated to bridge the innate immune system and atherosclerosis. NLRP3 is a member of the NLR family. Upon activation, it initiates IL-1β and IL-18 processing, a key step in the inflammatory process of atherosclerosis.

Material and methods: We used three different types of lipoproteins, ox-LDL, ox-HDL, and HDL, in Thp-1 at the concentration of 50 mg/l, 100 mg/l, and 150 mg/l respectively. Using real-time polymerase chain reaction and western blot, ELISA detected the expression of NLRP3 and downstream cytokines. NLRP3 siRNA was constructed to down-regulate expression of the NLRP3 gene via the RNA interference technique. 150 mg/l of ox-LDL, ox-HDL and HDL was added to the Thp-1 cell line respectively. We observed the changes in the expression of caspase-1, IL-1β and IL-18 when the NLRP3 gene was down-regulated.

Results: Ox-LDL and ox-HDL addition not only increases the expression of NLRP3, but also activates the NLRP3 downstream cytokines and caspase-1 and induces IL-1β and IL-18 secretion. Moreover, the effects of activation and induction are shown to have a dose-dependent manner. Expression of NLRP3 and its downstream inflammatory cytokines is reduced in the presence of HDL (p < 0.05). Furthermore, our data demonstrated that NLRP3 siRNA downregulates NLRP3 expression in mononuclear cells, thus leading to a dramatic reduction in the expression of caspase-1, IL-1β and IL-18 (p < 0.05).

Conclusions: The data suggest that activation of the NLRP3 inflammasome is a critical step in caspase-1 activation and IL-1β and IL-18 secretion. Interference with the NLRP3 inflammasome can significantly inhibit the generation of cytokines, thus impeding the pathogenesis of inflammation.

No MeSH data available.


NLRP3 siRNA interferes with expression of NLRP3 and caspase-1*p < 0.05, statistically significantly different compared to the group which did not transfect with NLRP3 siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5016581&req=5

Figure 0004: NLRP3 siRNA interferes with expression of NLRP3 and caspase-1*p < 0.05, statistically significantly different compared to the group which did not transfect with NLRP3 siRNA.

Mentions: We found that ox-LDL, ox-HDL and HDL have biological effects on the expression of NLRP3 and its downstream effectors (caspase-1, IL-1β and IL-18). This suggests that ox-LDL, ox-HDL and HDL are involved in the pathogenesis of atherosclerosis by affecting the function of the NLRP3 inflammasome. In this experiment, NLRP3 siRNA was constructed to down-regulate expression of the NLRP3 gene via the RNA interference technique. The data showed that expression of NLRP3 is significantly reduced by NLRP3 siRNA in the groups of ox-LDL, ox-HDL and HDL. On average, the level of reduction in the expression of NLRP3 is 60–70%. However, the inhibition levels among the three groups are not statistically significantly different (Figure 4 A). We also found that the level of caspase-1 expression is approximately 70% decreased in each group transfected with NLRP3 siRNA, and the inhibition level among groups is not statistically significantly different. The data clearly indicated that interference with NLRP3 can effectively inhibit the expression of caspase-1 (Figure 4 B).


NLRP3 inflammasome: a novel link between lipoproteins and atherosclerosis
NLRP3 siRNA interferes with expression of NLRP3 and caspase-1*p < 0.05, statistically significantly different compared to the group which did not transfect with NLRP3 siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5016581&req=5

Figure 0004: NLRP3 siRNA interferes with expression of NLRP3 and caspase-1*p < 0.05, statistically significantly different compared to the group which did not transfect with NLRP3 siRNA.
Mentions: We found that ox-LDL, ox-HDL and HDL have biological effects on the expression of NLRP3 and its downstream effectors (caspase-1, IL-1β and IL-18). This suggests that ox-LDL, ox-HDL and HDL are involved in the pathogenesis of atherosclerosis by affecting the function of the NLRP3 inflammasome. In this experiment, NLRP3 siRNA was constructed to down-regulate expression of the NLRP3 gene via the RNA interference technique. The data showed that expression of NLRP3 is significantly reduced by NLRP3 siRNA in the groups of ox-LDL, ox-HDL and HDL. On average, the level of reduction in the expression of NLRP3 is 60–70%. However, the inhibition levels among the three groups are not statistically significantly different (Figure 4 A). We also found that the level of caspase-1 expression is approximately 70% decreased in each group transfected with NLRP3 siRNA, and the inhibition level among groups is not statistically significantly different. The data clearly indicated that interference with NLRP3 can effectively inhibit the expression of caspase-1 (Figure 4 B).

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Pattern recognition receptor-mediated signaling pathways have recently been elucidated to bridge the innate immune system and atherosclerosis. NLRP3 is a member of the NLR family. Upon activation, it initiates IL-1&beta; and IL-18 processing, a key step in the inflammatory process of atherosclerosis.

Material and methods: We used three different types of lipoproteins, ox-LDL, ox-HDL, and HDL, in Thp-1 at the concentration of 50 mg/l, 100 mg/l, and 150 mg/l respectively. Using real-time polymerase chain reaction and western blot, ELISA detected the expression of NLRP3 and downstream cytokines. NLRP3 siRNA was constructed to down-regulate expression of the NLRP3 gene via the RNA interference technique. 150 mg/l of ox-LDL, ox-HDL and HDL was added to the Thp-1 cell line respectively. We observed the changes in the expression of caspase-1, IL-1&beta; and IL-18 when the NLRP3 gene was down-regulated.

Results: Ox-LDL and ox-HDL addition not only increases the expression of NLRP3, but also activates the NLRP3 downstream cytokines and caspase-1 and induces IL-1&beta; and IL-18 secretion. Moreover, the effects of activation and induction are shown to have a dose-dependent manner. Expression of NLRP3 and its downstream inflammatory cytokines is reduced in the presence of HDL (p &lt; 0.05). Furthermore, our data demonstrated that NLRP3 siRNA downregulates NLRP3 expression in mononuclear cells, thus leading to a dramatic reduction in the expression of caspase-1, IL-1&beta; and IL-18 (p &lt; 0.05).

Conclusions: The data suggest that activation of the NLRP3 inflammasome is a critical step in caspase-1 activation and IL-1&beta; and IL-18 secretion. Interference with the NLRP3 inflammasome can significantly inhibit the generation of cytokines, thus impeding the pathogenesis of inflammation.

No MeSH data available.