Page 7 - 南京医科大学学报自然科学版

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