基于Caspase-1研究NLRP3/Caspase-1通路对成骨细胞焦亡作用机制

doi:10.3969/j.issn.2095-378X.2023.03.002

外科研究与新技术 ›› 2023, Vol. 12 ›› Issue (3): 163-167.doi: 10.3969/j.issn.2095-378X.2023.03.002

基于Caspase-1研究NLRP3/Caspase-1通路对成骨细胞焦亡作用机制

于涛, 王海

牡丹江医学院附属第二医院骨外科, 黑龙江牡丹江 157000

收稿日期:2023-02-07 出版日期:2023-09-28 发布日期:2023-10-24
通讯作者: 王海,电子信箱：mswzhw@126.com
作者简介:于涛（1984—）,男,硕士研究生,主治医师,从事临床骨外科工作
基金资助:
黑龙江省卫生健康委科研课题（2020-397）

Study on the mechanism of NLRP3/Caspase-1 pathway on osteoblast pyroptosis based on Caspase-1

YU Tao, WANG Hai

Department of Orthopedic Surgery, The Second Hospital of Mudanjiang Medical College,Mudanjiang 157000, Heilongjiang, China

Received:2023-02-07 Online:2023-09-28 Published:2023-10-24

摘要/Abstract

摘要： 目的通过调控半胱氨酸蛋白酶-1（Caspase-1）水平探讨核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）/Caspase-1通路对成骨细胞焦亡的影响机制。方法培养小鼠胚胎MC3T3-E1成骨细胞,分为A组（高糖处理）、B组（高糖+Caspase-1抑制剂处理）、C组（高糖+Caspase-1 siRNA处理）和对照组。采用吸光法检测成骨细胞乳酸脱氢酶（LDH）释放量,采用Hoechst33342/PI染色法检测成骨细胞焦亡情况,分别采用RT-PCR技术和Western blot技术检测成骨细胞中NLRP3、Caspase-1、白细胞介素-1β（IL-1β）和IL-18相对表达水平。结果 A组、B组和C组MC3T3-E1成骨细胞焦亡比例和LDH活性均显著高于对照组,B组、C组焦亡比例和LDH活性显著低于A组（P<0.05）。A组、B组和C组NLRP3、Caspase-1、IL-1β、IL-18 mRNA和蛋白相对表达水平均显著高于对照组,B组、C组Caspase-1、IL-1β、IL-18 mRNA和蛋白相对表达水平显著低于A组（P<0.05）。A组、B组和C组NLRP3 mRNA和蛋白相对表达水平比较差异无统计学意义（P>0.05）。结论通过抑制Caspase-1能够抑制高糖引起的成骨细胞NLRP3升高导致的IL-1β、IL-18升高,进而抑制成骨细胞焦亡作用。

关键词: 成骨细胞, 细胞焦亡, Caspase-1, NLRP3/Caspase-1通路, Caspase-1抑制剂, NLRP3炎性小体

Abstract: Objective To explore the mechanism of nucleotide binding oligomerization domain-like receptor protein 3 （NLRP3）/cysteine aspartate specific proteinase-1（Caspase-1） pathway on osteoblast pyroptosis by regulating the level of Caspase-1. Methods Mouse embryonic MC3T3-E1 osteoblasts were cultured and divided into group A with high sugar treatment, group B with high sugar + Caspase-1 inhibitor treatment, group C with high sugar + Caspase-1 siRNA treatment, and control group. The level of lactate dehydrogenase （LDH） released from osteoblasts was detected by light absorption method, the osteoblast pyroptosis was detected by Hoechst33342/PI staining, and the NLRP3, Caspase-1, interleukin-1β （IL-1β）, and IL-18 of osteoblasts were detected by RT-PCR and Western blot respectively. Results The pyroptosis ratio and LDH activity of MC3T3-E1 osteoblasts in group A, B, and C were significantly higher than those in the control group, while those in group B and C were significantly lower than those in group A （P<0.05）. The relative expression levels of NLRP3, Caspase-1, IL-1 β, and IL-18 mRNA and protein in group A, B, and C were higher than those in the control group, while the relative expression levels of Caspase-1, IL-1 β, and IL-18 mRNA and protein in group B and C were significantly lower than those in group A （P<0.05）. There was no significant difference in the relative expression levels of NLRP3 mRNA and protein among group A, B, and C （P>0.05）. Conclusion Inhibition of Caspase-1 inhibits the elevation of IL-1β and IL-18 caused by the elevation of NLRP3 in osteoblasts due to high glucose, thus inhibiting osteoblast pyroptosis.

Key words: Osteoblast, Pyroptosis, Caspase-1, NLRP3/Caspase-1 pathway, Caspase-1 inhibitor, NLRP3 inflammasome

中图分类号:

R329.25

于涛, 王海. 基于Caspase-1研究NLRP3/Caspase-1通路对成骨细胞焦亡作用机制[J]. 外科研究与新技术, 2023, 12(3): 163-167.

YU Tao, WANG Hai. Study on the mechanism of NLRP3/Caspase-1 pathway on osteoblast pyroptosis based on Caspase-1[J]. Surgical Research and New Technique, 2023, 12(3): 163-167.

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基于Caspase-1研究NLRP3/Caspase-1通路对成骨细胞焦亡作用机制

Study on the mechanism of NLRP3/Caspase-1 pathway on osteoblast pyroptosis based on Caspase-1

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