N-乙酰半胱氨酸对尼古丁诱导的MC3T3-E1细胞凋亡的作用及其机制

doi:10.13481/j.1671-587X.20220104

Abstract

Abstract: Objective

To investigate the effect of N-acetylcysteine （NAC） on the apoptosis of mouse MC3T3-E1 preosteoblast cells induced by nicotine， and to clarify its mechanism.

Methods

The MC3T3-E1 cells were cultured in vitro and treated with different concentrations （0， 1.0， 2.5， 5.0 and 7.5 mmol·L^－1） of nicotine to induce the MC3T3-E1 cell injury； MTT method was used to determine the proliferation activities of cells in various groups and the half inhibitory concentration（IC₅₀）of nicotine was determined.The MC3T3-E1 cells were divided into control group， nicotine group， NAC group and nicotine+different concentrations（2.5，5.0，and 7.5 mmol·L^－1） NAC groups according the IC₅₀values， and MTT method was used to determine the proliferation activities of cells in various groups； the optimun concentration of NAC was confirmed.The cells were divided into control group，nicotine （4.25 mmol·L^－1） group，NAC（5.0 mmol·L^－1） group and nicotine（4.25 mmol·L^－1） +NAC（5.0 mmol·L^－1） group.The levels of mitochondrial ROS （mitoROS） was assessed with MitoSOX fluorescence staining method； Flow cytometry was used to detect the apoptic rates of MC3T3-E1 cells in various groups； Western blotting method was used to detect the expression level of apoptosis-related proteins B-cell lymphoma-2 （Bcl-2） and Bcl-2 associated X proteins （Bax）in the cells in various groups， and the ratios of Bax/Bcl-2 was calculated.

Results

Compared with 0 mmol·L^－1 nicotine group， the proliferation activities of MC3T3-E1 cells in different concentrations of nicotine groups were significantly decreased （P<0.05） in a dose-dependent manner. The IC₅₀ of nicotine was （4.25±0.03） mmol·L^－1.Compared with control group，the proliferation activity of MC3T3-E1 cells in 4.25 mmol·L^－1 nicotine group was significantly decreased （P<0.05）； compared with nicotine group， the proliferation activites of MC3T3-E1 cells in nicotine+different concentrations of NAC groups were increased（P<0.05）；the proliferation activity of cells in nicotine+5.0 mmol·L^－1NAC group was the highest. Compared with control group， the levels of mitoROS in the cells in nicotine group was significantly increased （P<0.05）， the apoptotic rate of cells was significantly increased （P<0.05），and the ratio of Bax/Bcl-2 was increased （P<0.05）；compared with nicotine group， the levels of mitoROS in the cells in NAC and nicotine+NAC group were significantly decreased （P<0.05）， the apoptotic rate was significantly reduced （P<0.05），and the ratio of Bax/Bcl-2 in nicotine+NAC group was reduced （P<0.05）.

Conclusion

NAC can alleviate the apoptosis of MC3T3-E1 cells induced by nicotine， and its mechanism may be related to the mitochondrial oxidative stress.

Key words: MC3T3-E1 cells, Nicotine, N-acetylcysteine, Apoptosis, Oxidative stress

CLC Number:

Leihua CUI,Yubo HOU,Chang SU,Minghe LI,Xin NIE. Effect of N-acetylcysteine on apoptosis of MC3T3-E1 cells induced by nicotine and its mechanism[J].Journal of Jilin University(Medicine Edition), 2022, 48(1): 26-32.

Figures/Tables 5

References 26

1	HAJISHENGALLIS G， CHAVAKIS T. Local and systemic mechanisms linking periodontal disease and inflammatory comorbidities［J］. Nat Rev Immunol， 2021， 21（7）： 426-440.
2	ALHARTHI S S Y， NATTO Z S， MIDLE J B， et al. Association between time since quitting smoking and periodontitis in former smokers in the National Health and Nutrition Examination Surveys （NHANES） 2009 to 2012［J］. J Periodontol， 2019， 90（1）： 16-25.
3	CHOI H K， ATAUCURI-VARGAS J， LIN C， et al. The current state of tobacco cessation treatment［J］. Cleve Clin J Med， 2021， 88（7）： 393-404.
4	TARANTINO U， CARIATI I， GREGGI C， et al. Skeletal system biology and smoke damage： from basic science to medical clinic［J］.Int J Mol Sci，2021，22（12）： 6629.
5	ZHOU Z， LIU F， WANG L， et al. Inflammation has synergistic effect with nicotine in periodontitis by up-regulating the expression of α7 nAChR via phosphorylated GSK-3β［J］. J Cell Mol Med， 2020， 24（4）： 2663-2676.
6	SILVA H. Tobacco use and periodontal disease—the role of microvascular dysfunction［J］. Biology， 2021， 10（5）： 441.
7	CHEN M， CAI W， ZHAO S， et al. Oxidative stress-related biomarkers in saliva and gingival crevicular fluid associated with chronic periodontitis： a systematic review and meta-analysis［J］. J Clin Periodontol， 2019， 46（6）： 608-622.
8	SHIH Y M， CHANG Y J， COOKE M S， et al. Alkylating and oxidative stresses in smoking and non-smoking patients with COPD： Implications for lung carcinogenesis［J］. Free Radic Biol Med， 2021， 164： 99-106.
9	MURPHY M P， HARTLEY R C. Mitochondria as a therapeutic target for common pathologies［J］. Nat Rev Drug Discov， 2018， 17（12）： 865-886.
10	JIA G， MENG Z， LIU C， et al. Nicotine induces cardiac toxicity through blocking mitophagic clearance in young adult rat［J］. Life Sci， 2020， 257： 118084.
11	GODOY J A， VALDIVIESO A G， INESTROSA N C. Nicotine modulates mitochondrial dynamics in hippocampal neurons［J］. Mol Neurobiol， 2018，55（12）： 8965-8977.
12	HE Y T， GAN X Q， ZHANG L， et al. CoCl₂ induces apoptosis via a ROS-dependent pathway and Drp1-mediated mitochondria fission in periodontal ligament stem cells［J］. Am J Physiol Cell Physiol， 2018， 315（3）： C389-C397.
13	HUANG C， SHI G. Smoking and microbiome in oral， airway， gut and some systemic diseases［J］. J Transl Med， 2019， 17（1）： 225.
14	LEITE F R M， NASCIMENTO G G， SCHEUTZ F， et al. Effect of smoking on periodontitis： a systematic review and meta-regression［J］. Am J Prev Med， 2018， 54（6）： 831-841.
15	KUMAR P S. Interventions to prevent periodontal disease in tobacco-， alcohol-， and drug-dependent individuals［J］. Periodontol 2000， 2020， 84（1）： 84-101.
16	CHAFFEE B W， PERSAI D， VORA M V. Interdental cleaning and oral health status in an adult cohort， 2015 to 2018［J］. J Dent Res， 2020， 99（10）： 1150-1156.
17	LIANG D， WANG K J， TANG Z Q， et al. Effects of nicotine on the metabolism and gene expression profile of Sprague‑Dawley rat primary osteoblasts［J］. Mol Med Rep， 2018， 17（6）： 8269-8281.
18	KUNTIC M， OELZE M， STEVEN S， et al. Short-term e-cigarette vapour exposure causes vascular oxidative stress and dysfunction： evidence for a close connection to brain damage and a key role of the phagocytic NADPH oxidase （NOX-2）［J］. Eur Heart J， 2020， 41（26）： 2472-2483.
19	HOLLIDAY R， CHAFFEE B W， JAKUBOVICS N S， et al. Electronic cigarettes and oral health［J］. J Dent Res， 2021， 100（9）： 906-913.
20	HOLLIDAY R S， CAMPBELL J， PRESHAW P M. Effect of nicotine on human gingival， periodontal ligament and oral epithelial cells. A systematic review of the literature［J］. J Dent， 2019， 86： 81-88.
21	KIM W S， LEE K S， KIM J H， et al. The caspase-8/Bid/cytochrome c axis links signals from death receptors to mitochondrial reactive oxygen species production［J］. Free Radic Biol Med， 2017， 112： 567-577.
22	KHAN N A， LAWYER G， MCDONOUGH S， et al. Systemic biomarkers of inflammation， oxidative stress and tissue injury and repair among waterpipe， cigarette and dual tobacco smokers［J］.Tob Control，2020，29（）： s102-s109.
23	ABATE M， FESTA A， FALCO M， et al. Mitochondria as playmakers of apoptosis， autophagy and senescence［J］. Semin Cell Dev Biol， 2020， 98： 139-153.
24	MARINUCCI L， BALLONI S， FETTUCCIARI K， et al. Nicotine induces apoptosis in human osteoblasts via a novel mechanism driven by H₂O₂ and entailing Glyoxalase 1-dependent MG-H1 accumulation leading to TG2-mediated NF-kB desensitization： Implication for smokers-related osteoporosis［J］. Free Radic Biol Med， 2018， 117： 6-17.
25	LAHMOUZR J， SIMAIN-SATO F， DEFRESNE M P，et al. Effect of nicotine on rat gingival fibroblasts in vitro ［J］. Connect Tissue Res， 2000， 41（1）： 69-80.
26	BAI X， CHEN S， XU K， et al. N-acetylcysteine combined with dexamethasone treatment improves sudden sensorineural hearing loss and attenuates hair cell death caused by ROS stress［J］. Front Cell Dev Biol， 2021， 9： 659486.

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Effect of N-acetylcysteine on apoptosis of MC3T3-E1 cells induced by nicotine and its mechanism

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