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第43卷第12期 南京医科大学学报(自然科学版)
2023年12月 Journal of Nanjing Medical University(Natural Sciences) ·1623 ·
·基础研究·
TLR2/NF⁃κB通路介导MPP 诱导的RSC96细胞凋亡与自噬
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李杨夏 ,佟 晴 ,程 越 ,耿 铫 ,王田田 ,张克忠 1*
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南京医科大学第一附属医院神经内科,江苏 南京 210029;南京医科大学基础医学院药理学系,江苏 南京 211166
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[摘 要] 目的:探讨1⁃甲基⁃4⁃苯基⁃吡啶离子(1⁃methyl⁃4⁃phenyl⁃pyridinium,MPP )诱导的RSC96细胞凋亡与自噬功能障碍
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与TLR2/NF⁃κB信号通路的关系。方法:将RSC96细胞分为PBS组、MPP 组和MPP +CU⁃CPT22组;CCK⁃8检测不同MPP 浓度
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(0.1、0.3、0.5、0.7、0.9 mmol/L)处理后细胞存活率;TUNEL检测细胞凋亡水平;RT⁃qPCR检测Toll样受体2(TLR2)mRNA转录水
平;Western blot检测凋亡相关指标Bcl⁃2/Bax、cleaved caspase⁃3/caspase⁃3,自噬相关指标LC3Ⅱ/LC3Ⅰ和P62,以及TLR2、p⁃NF⁃
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κB/NF⁃κB蛋白表达水平。结果:与PBS组相比,MPP 组细胞存活率下降且呈浓度依赖性,TUNEL染色阳性细胞数增多,Bcl⁃2/
Bax蛋白比值水平下降,cleaved caspase⁃3/caspase⁃3比值增高。同时LC3Ⅱ/LC3Ⅰ比值下降,P62表达增加,p⁃NF⁃κB/NF⁃κB比
值表达增加。RT⁃qPCR和Western blot结果显示,MPP 上调TLR2的表达。此外,与MPP 组相比,MPP +CU⁃CPT22组TUNEL阳性
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细胞数目减少,Bcl⁃2/Bax比值升高,cleaved caspase⁃3/caspase⁃3比值降低;同时,LC3Ⅱ/LC3Ⅰ比值上升,P62水平下降。结论:
MPP 刺激诱导RSC96细胞凋亡和自噬水平失衡,发生机制可能与TLR2/NF⁃κB通路的激活有关。
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[关键词] 帕金森病;雪旺细胞;Toll样受体2;自噬;凋亡
[中图分类号] R285.5 [文献标志码] A [文章编号] 1007⁃4368(2023)12⁃1623⁃07
doi:10.7655/NYDXBNS20231201
TLR2/NF⁃κB pathway mediates MPP ⁃induced apoptosis and autophagy in RSC96 cells
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LI Yangxia ,TONG Qing ,CHENG Yue ,GENG Yao ,WANG Tiantian ,ZHANG Kezhong 1*
Department of Neurology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;Department
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of Pharmacology,School of Basic Medical Sciences,Nanjing Medical University,Nanjing 211166,China
[Abstract] Objective:To investigate the relationship between TLR2/NF⁃κB signaling pathway and the apoptosis and autophagy
dysfunction of RSC96 cells induced by 1⁃methyl⁃4⁃phenyl⁃pyridinium(MPP). Methods:RSC96 cells were divided into the PBS group,
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MPP group,and MPP + CU ⁃ CPT22 group. Cell survival rate was detected using CCK ⁃ 8 after treatment with different MPP +
concentrations(0.1,0.3,0.5,0.7,0.9 mmol/L). Cell apoptosis was detected by TUNEL staining. RT⁃qPCR was performed to detect the
TLR2 mRNA level. Western blot was performed to detect the expression levels of apoptosis related indicators Bcl ⁃ 2/Bax,cleaved
caspase⁃3/caspase⁃3,autophagy⁃related indicators LC3II/LC3I and P62,as well as TLR2 and p⁃NF⁃κB/NF⁃κB. Results:Compared
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with the PBS group,the cell viability of the MPP group decreased in a concentration⁃dependent manner,the number of TUNEL⁃
staining positive cells increased,the ratio of Bcl⁃2/Bax decreased while cleaved caspase⁃3/caspase⁃3 ratio increased,as well as had a
decrease in the ratio of LC3Ⅱ/LC3Ⅰ and an increase in P62 and p⁃NF⁃κB/NF⁃κB ratio elvel. RT⁃qPCR and Western blot results
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showed that MPP upregulated the expression of TLR2. In addition,compared with the MPP group,the MPP +CU⁃CPT22 group
showed a decrease in the number of TUNEL⁃staining positive cells,an increase in Bcl⁃2/Bax level,and a decrease in cleaved caspase⁃3/
caspase ⁃ 3 ratio. Meanwhile,LC3 Ⅱ/LC3 Ⅰ ratio was increased,and the P62 expression level was decreased. Conclusion:MPP +
stimulation induced apoptosis and the imbalance of autophagy in RSC96 cells,and the mechanism may be related to the activation of
the TLR2/NF⁃κB signaling pathway.
[Key words] Parkinson’s disease;Schwann cells;Toll⁃like receptor 2;autophagy;apoptosis
[J Nanjing Med Univ,2023,43(12):1623⁃1629]
[基金项目] 国家自然科学基金(8207051941)
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通信作者(Corresponding author),E⁃mail:kezhong_zhang1969@126.com