Reactive oxygen species/c-Jun N-terminal kinase/nuclear factor kappa-B signaling molecules are involved in pe-riodontitis-induced liver injury by regulating apoptosis

doi:10.7518/hxkq.2022.05.005

Abstract

Abstract:

Objective The occurrence and development of periodontitis and nonalcoholic fatty liver disease (NAFLD) are closely related to the accumulation of reactive oxygen species (ROS). ROS are involved in regulating the activation of c-Jun N-terminal kinase (JNK)/nuclear factor kappa-B (NF-κB) signaling molecules. When the signaling molecules are overactivated by ROS, the internal environment of the body can be disturbed. Therefore, this study aimed to explore the mechanism by which ROS/JNK/NF-κB signaling molecules are involved in periodontitis-induced liver injury. Methods Twelve SPF male Wistar rats were randomly divided into control and periodontitis groups. The perio-dontitis model of rats was established by wire ligation in the neck of bilateral maxillary first molars. After 8 weeks, the periodontal clinical indexes of the rats were examined, and the rats were sacrificed. Micro-CT reconstruction of a three-dimensional alveolar bone structure and analysis of alveolar bone absorption were conducted. Pathological changes in the periodontal and liver tissues were analyzed by histopathology. MitoSOX red reagent was used to detect the ROS content in liver tissue. Biochemical kits were used to detect liver function and oxidative stress biomarkers. The mRNA expression levels ofinterleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), NF-κB, BCL2-associated X (Bax), and B-cell lymphoma-2 (Bcl-2) in liver tissue were detected through quantitative real-time polymerase chain reaction (qRT-PCR). The protein expression levels of phosphorylated c-Jun N-terminal kinase (P-JNK), JNK, NF-κB, Caspase-3, Bax, and Bcl-2 in liver tissue were detected by Western blot. Apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Results Micro-CT results showed that the mice in the periodontitis group had obvious alveolar bone resorption and significantly greater distance from the cemento-enamel junction to the alveolar bone crest than those in the control group. Histopathological results showed that a large number of inflammatory cells were infiltrated in the periodontal tissue of the periodontitis group. In addition, the resorption of alveolar ridge bone was obvious and liver tissue structure was destroyed, with balloon-like changes and red lipid droplets. MitoSOX red staining results showed that the ROS level was significantly higher in the liver tissue of the periodontitis group than in that of the control group. Biochemical test results showed that the aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in the serum of the periodontitis group were higher than those in the serum of the control group. The levels of superoxide dismutase (SOD) and glutathione (GSH) in liver tissue decreased, whereas the that of malondialdehyde (MDA) increased. Western blot and qRT-PCR results revealed that the mRNA levels of IL-6, TNF-α, Bax, and NF-κB and the protein levels of P-JNK/JNK, NF-κB, Caspase-3, and Bax were significantly higher in the liver tissue of the perio-dontitis group than in that of the control group. Meanwhile, the mRNA and protein levels of Bcl-2 were lower in the periodontitis group than in the control group. TUNEL staining showed that the number of apoptotic cells was significantly higher in the periodontitis group than in the control group. Conclusion ROS/JNK/NF-κB signaling molecules are involved in periodontitis-induced liver injury by regulating apoptosis.

Key words: periodontitis, liver injury, oxidative stress, c-Jun N-terminal kinases, nuclear factor kappa-B, apoptosis

CLC Number:

Q 257

Cao Niuben, Liu Xiaomeng, Deng Yu, Liu Xinchan, Xin Yu, Yu Weixian. Reactive oxygen species/c-Jun N-terminal kinase/nuclear factor kappa-B signaling molecules are involved in pe-riodontitis-induced liver injury by regulating apoptosis[J]. West China Journal of Stomatology, 2022, 40(5): 532-540.

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基因	引物序列（5’—3’）	碱基数/bp	基因号
IL-6^[14]	F：ATTGTATGAACAGCGATGATGCAC	20	NM_012489.2
	R：CCAGGTAGAAACGGAACTCCAGA	20
TNF-α^[14]	F：GGCGTGTTCATCCGTTCTC	24	NM_012675.3
	R：CTTCAGCGTCTCGTGTGTTTCT	23
NF-κB^[14]	F：CGACGTATTGCTGTGCCTTC	19	NM_199267.2
	R：TTGAGATCTGCCCAGGTGGTA	22
Bax^[15]	F：CCACCAAGAAGCTGAGCGA	19	NM_017059.2
	R：GCTGCCACACGGAAGAAGA	19
Bcl-2^[15]	F：CTCTGTGGATGACTGAGTACCTG	23	NM_016993.2
	R：GAGCAGCGTCTTCAGAGACAG	21
β-actin^[14]	F：GGAGATTACTGCCCTGGCTCCTA	23	NM_031144.3
	R：GACTCATCGTACTCCTGCTTGCTG	24

牙周临床指标	对照组	牙周炎组
PD/mm	0.21±0.03	2.26±0.08***
BI	0.20±0.20	2.60±0.25***
TM	0.00±0.00	1.00±0.32*

生化指标	对照组	牙周炎组
SOD/（U·mgprot^-1）	83.27±3.72	67.10±2.22*
GSH/（μmol·gprot^-1）	63.78±3.22	31.26±5.34**
MDA/（nmol·mgprot^-1）	1.56±0.08	2.14±0.13*
AST/（U·L^-1）	36.00±1.29	63.57±1.94***
ALT/（U·L^-1）	34.16±0.51	50.79±1.38***

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