慢性牙周炎通过PERK-CHOP信号通路介导睾周脂肪细胞凋亡的研究

doi:10.13591/j.cnki.kqyx.2022.10.003

摘要/Abstract

摘要： 目的探讨慢性牙周炎通过蛋白激酶R样内质网激酶(protein kinase R-like ER kinase,PERK)-C/EBP同源蛋白(C/EBP homologous protein,CHOP)信号通路对大鼠睾周脂肪细胞凋亡的影响。方法将20只雄性SD大鼠随机分为实验组和对照组。实验组构建大鼠慢性牙周炎模型,对双侧上颌第二磨牙行丝线结扎并龈沟内注射牙龈卟啉单胞菌脂多糖(Porphyromonas gingivalis-lipopolysaccharid,P. gingivalis-LPS);对照组大鼠龈沟内注射等量PBS。建模3个月后处死大鼠,通过测量牙龈出血指数(gingival bleeding index,GBI)及牙周袋探诊深度(probing pocket depth,PPD)评价牙周组织炎症,采用Micro-CT分析大鼠第二磨牙牙槽骨吸收程度。原位细胞凋亡检测法(TUNEL)染色观察睾周脂肪细胞凋亡情况。蛋白质印迹法(Western blot)检测PERK、p-PERK、CHOP、Caspase-12、Cleaved-Caspase-3的蛋白表达水平。结果实验组大鼠GBI、PPD及釉牙骨质界至牙槽嵴顶距离显著高于对照组(P<0.05),出现明显牙龈软组织炎症和牙槽骨破坏吸收,大鼠慢性牙周炎模型建立成功。实验组大鼠睾周脂肪细胞凋亡率显著增加(P<0.05),p-PERK、CHOP、Caspase-12、Cleaved-Caspase-3蛋白表达水平显著升高(P<0.05)。结论慢性牙周炎通过PERK-CHOP信号通路介导大鼠睾周脂肪细胞的凋亡。

关键词: 牙周炎, 蛋白激酶R样内质网激酶, C/EBP同源蛋白, 睾周脂肪细胞, 凋亡

Abstract: Objective To investigate the effect of chronic periodontitis on apoptosis of rat peritesticular adipocytes through protein kinase R-like ER kinase (PERK)-C/EBP homologous protein (CHOP) signal pathway. Methods Twenty male SD rats were randomly divided into experimental group and control group. Experimental group rat model of chronic periodontitis was established. The bilateral maxillary second molars were ligated with silk thread and injected with Porphyromonas gingivalis-lipopolysaccharide (P. gingivalis-LPS) into the gingival sulcus. The control group was injected with the same amount of PBS into the gingival sulcus. After 3 months of modeling, the rats were killed. The periodontal inflammation was evaluated by measuring the gingival bleeding index (GBI) and the probing pocket depth (PPD). The alveolar bone resorption of the second molar was analyzed by Micro-CT. The apoptosis of peritesticular adipocyte was observed by TUNEL staining. Western blot was used to detect the protein expression levels of PERK, p-PERK, CHOP, Caspase-12 and Cleaved-Caspase-3. Results GBI, PPD and CEJ-ABC in the experimental group were significantly higher than those in the control group (P<0.05), and the rat model of chronic periodontitis was successfully established. The apoptosis rate of peritesticular adipocytes in the experimental group increased significantly (P<0.05), and the expression levels of p-PERK, CHOP, Caspase-12 and Cleaved-Caspase-3 protein increased significantly (P<0.05). Conclusion Chronic periodontitis mediates apoptosis of peritesticular adipocyte in rats through the PERK-CHOP signaling pathway.

Key words: periodontitis, protein kinase R-like ER kinase, C/EBP homologous protein, peritesticular adipocytes, apoptosis

中图分类号:

R781.4

陈伊燕, 伍倩琪, 陆佳艺, 郭吕华. 慢性牙周炎通过PERK-CHOP信号通路介导睾周脂肪细胞凋亡的研究[J]. 口腔医学, 2022, 42(10): 878-882.

CHEN Yiyan, WU Qianqi, LU Jiayi, GUO Lyuhua. Study on peritesticular adipocytes apoptosis mediated by PERK-CHOP signaling pathway in chronic periodontitis[J]. Stomatology, 2022, 42(10): 878-882.

参考文献

[1] Pirih FQ, Monajemzadeh S, Singh N, et al. Association between metabolic syndrome and periodontitis:The role of lipids, inflammatory cytokines, altered host response, and the microbiome[J]. Periodontol 2000, 2021, 87(1):50-75.
[2] Jepsen S, Suvan J, Deschner J. The association of periodontal diseases with metabolic syndrome and obesity[J]. Periodontol 2000, 2020, 83(1):125-153.
[3] 郑旭, 甘姗灵, 郭竹玲. 牙周基础治疗调节2型糖尿病患者代谢的研究进展[J]. 口腔医学, 2019, 39(2):183-187.
[4] Li SQ, Gao HX, Hasegawa Y, et al. Fight against fibrosis in adipose tissue remodeling[J]. Am J Physiol Endocrinol Metab, 2021, 321(1):E169-E175.
[5] Reyes M, González L, Ibeas K, et al. White adipose tissue-infiltrated CD11b⁺ myeloid cells are a source of S100A4, a new potential marker of hepatic damage[J]. Eur J Endocrinol, 2021, 184(4):533-541.
[6] 陆佳艺, 伍倩琪, 陈伊燕, 等. 牙龈卟啉单胞菌来源的脂多糖通过X盒结合蛋白1调控脂肪细胞胰岛素信号通路的机制研究[J]. 华西口腔医学杂志, 2022, 40(2):148-154.
[7] Iurlaro R, Muñoz-Pinedo C. Cell death induced by endoplasmic Reticulum stress[J]. FEBS J, 2016, 283(14):2640-2652.
[8] Liu R, Li N, Liu N, et al. Effects of systemic ornidazole, systemic and local compound ornidazole and pefloxacin mesylate on experimental periodontitis in rats[J]. Med Sci Monit, 2012, 18(3):BR95-BR102.
[9] Soliman H, Varela JN, Nyamandi V, et al. Attenuation of obesity-induced insulin resistance in mice with heterozygous deletion of ROCK2[J]. Int J Obes (Lond), 2016, 40(9):1435-1443.
[10] Kitada Y, Kajita K, Taguchi K, et al. Blockade of sphingosine 1-phosphate receptor 2 signaling attenuates high-fat diet-induced adipocyte hypertrophy and systemic glucose intolerance in mice[J]. Endocrinology, 2016, 157(5):1839-1851.
[11] 尹忠浩, 朱载瓯, 杨月美, 等. 右美托咪定对大鼠肋间后动脉穿支皮瓣缺血再灌注损伤保护作用的初步研究[J]. 口腔医学, 2022, 42(5):391-398.
[12] Chang LY, Lai CH, Kuo CH, et al. Recombinant thrombomodulin lectin-like domain attenuates Porphyromonas gingivalis lipopolysaccharide-induced osteoclastogenesis and periodontal bone resorption[J]. J Periodontol, 2021, 92(11):1622-1634.
[13] Kajiwara K, Sawa Y, Fujita T, et al. Immunohistochemical study for the expression of leukocyte adhesion molecules, and FGF23 and ACE2 in P. gingivalis LPS-induced diabetic nephropathy[J]. BMC Nephrol, 2021, 22(1):3.
[14] Bregaint S, Boyer E, Fong SB, et al. Porphyromonas gingivalis outside the oral cavity[J]. Odontology, 2022, 110(1):1-19.
[15] Chen TS, Kuo CH, Battsengel S, et al. Adipose-derived stem cells decrease cardiomyocyte damage induced by Porphyromonas gingivalis endotoxin through suppressing hypertrophy, apoptosis, fibrosis, and MAPK markers[J]. Environ Toxicol, 2018, 33(4):508-513.
[16] Kawamura N, Ohnuki Y, Matsuo I, et al. Effects of chronic Porphyromonas gingivalis lipopolysaccharide infusion on skeletal muscles in mice[J]. J Physiol Sci, 2019, 69(3):503-511.
[17] Bugueno IM, Batool F, Korah L, et al. Porphyromonas gingivalis differentially modulates apoptosome apoptotic peptidase activating factor 1 in epithelial cells and fibroblasts[J]. Am J Pathol, 2018, 188(2):404-416.
[18] Lv YT, Zeng JJ, Lu JY, et al. Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) influences adipocytes injuries through triggering XBP1 and activating mitochondria-mediated apoptosis[J]. Adipocyte, 2021, 10(1):28-37.
[19] 任庆源, 何武林, 王庆, 等. 持续静压力作用下内质网应激对牙周膜细胞成骨分化的影响[J]. 口腔疾病防治, 2019, 27(8):485-489.
[20] Xu J, Xiong M, Huang B, et al. Advanced glycation end products upregulate the endoplasmic Reticulum stress in human periodontal ligament cells[J]. J Periodontol, 2015, 86(3):440-447.
[21] Tan J, Zhou Y, Luo J, et al. High glucose inhibits the osteogenic differentiation of periodontal ligament stem cells in periodontitis by activating endoplasmic Reticulum stress[J]. Ann Transl Med, 2022, 10(4):204.
[22] 李鑫, 李艳, 丁旭, 等. 内质网应激在牙周炎影响全身疾病过程中的作用[J]. 国际口腔医学杂志, 2021, 48(1):12-17.
[23] Wu YT, Yue Y, Fu S, et al. Icariside Ⅱ prevents hypertensive heart disease by alleviating endoplasmic Reticulum stress via the PERK/ATF-4/CHOP signalling pathway in spontaneously hypertensive rats[J]. J Pharm Pharmacol, 2019, 71(3):400-407.
[24] Bai YD, Wei Y, Wu L, et al. C/EBP β mediates endoplasmic Reticulum stress regulated inflammatory response and extracellular matrix degradation in LPS-stimulated human periodontal ligament cells[J]. Int J Mol Sci, 2016, 17(3):385.
[25] Wang QX, Yuan XQ, Chen Y, et al. Endoplasmic Reticulum stress mediated MDRV p10.8 protein-induced cell cycle arrest and apoptosis through the PERK/eIF2α pathway[J]. Front Microbiol, 2018, 9:1327.
[26] de la Cadena SG, Hernández-Fonseca K, Camacho-Arroyo I, et al. Glucose deprivation induces Reticulum stress by the PERK pathway and caspase-7- and calpain-mediated caspase-12 activation[J]. Apoptosis, 2014, 19(3):414-427.
[27] Kamarehei M, Kabudanian Ardestani S, Firouzi M, et al. Increased expression of endoplasmic Reticulum stress-related caspase-12 and CHOP in the Hippocampus of EAE mice[J]. Brain Res Bull, 2019, 147:174-182.
[28] Huang H, An Y, Jiao WY, et al. CHOP/caspase-3 signal pathway involves in mitigative effect of selenium on lead-induced apoptosis via endoplasmic Reticulum pathway in chicken testes[J]. Environ Sci Pollut Res Int, 2018, 25(19):18838-18845.
[29] He BM, Zhang WX, Qiao JF, et al. Melatonin protects against COPD by attenuating apoptosis and endoplasmic Reticulum stress via upregulating SIRT1 expression in rats[J]. Can J Physiol Pharmacol, 2019, 97(5):386-391.
[30] Wongkrasant P, Pongkorpsakol P, Chitwattananont S, et al. Fructo-oligosaccharides alleviate inflammation-associated apoptosis of GLP-1 secreting L cells via inhibition of iNOS and Cleaved caspase-3 expression[J]. J Pharmacol Sci, 2020, 143(2):65-73.