沉默信息调节因子1对慢性牙周炎模型大鼠肾损伤的影响

doi:10.13481/j.1671-587X.20220513

摘要/Abstract

摘要：

目的探讨沉默信息调节因子1（SIRT1）在结扎诱导的慢性牙周炎模型大鼠肾损伤中的变化，阐明SIRT1对慢性牙周炎模型大鼠肾损伤的影响。方法 20只大鼠随机分为对照组和牙周炎组，每组10只。采用结扎双侧上颌第一磨牙颈部建立大鼠牙周炎模型。肉眼观察2组大鼠牙龈色形质，检测并记录牙龈出血指数（BI）、牙周探诊深度（PD）和牙齿松动度（TM），苏木素-伊红（HE）染色观察2组大鼠牙周组织病理形态表现，微型计算机断层成像（Micro-CT）评价2组大鼠牙槽骨吸收情况，HE和PAS染色观察2组大鼠肾组织病理形态表现，生化试剂盒检测2组大鼠肾脏功能指标和氧化应激水平，Western blotting法检测2组大鼠肾组织中肿瘤坏死因子α（TNF-α）和核因子κB（NF-κB） p65蛋白表达水平，实时荧光定量PCR（RT-qPCR）和免疫组织化学法检测2组大鼠肾组织中SIRT1和过氧化物酶体增殖物激活受体γ共激活因子1α（PGC-1α）mRNA及蛋白表达水平。结果牙周组织临床指标，与对照组比较，牙周炎组大鼠牙龈肿胀，质地松软，可探及深牙周袋，探诊易出血，牙齿出现不同程度松动。牙周组织HE染色和Micro-CT观察，与对照组比较，牙周炎组大鼠出现附着丧失，牙槽骨吸收。肾组织HE和PAS染色观察，与对照组比较，牙周炎组大鼠肾组织中肾小囊腔扩张，肾小球基底膜轻度增厚，肾小管上皮细胞脱落，刷状缘破坏。生化检测，2组大鼠肾脏功能指标比较差异无统计学意义（P>0.05）；与对照组比较，牙周炎组大鼠肾组织中总抗氧化状态（TAS）、锰-超氧化物歧化酶（Mn-SOD）和谷胱甘肽（GSH）水平降低（P<0.01），丙二醛（MDA）水平升高（P<0.01）。Western blotting法检测，与对照组比较，牙周炎组大鼠肾组织中TNF-α和NF-κB p65蛋白表达水平升高（P<0.05或P<0.01）。RT-qPCR和免疫组织化学法检测，与对照组比较，牙周炎组大鼠肾组织中SIRT1和PGC-1α mRNA及蛋白表达水平降低（P<0.05或P<0.01）。结论慢性牙周炎模型大鼠肾组织中SIRT1表达下调，氧化应激水平升高，加重了慢性牙周炎大鼠肾组织损伤。

关键词: 慢性牙周炎, 肾损伤, 沉默信息调节因子1, 大鼠

Abstract:

Objective To investigate the changes of silencing information regulator 1 （SIRT1） in ligation-induced kidney injury in the chronic periodontitis rats， and to clarify the effect of SIRT1 on kidney injury in the chronic periodontitis rats. Methods Twenty rats were randomly divided into control group and periodontitis group，with 10 rats in each group.The periodontitis model was established by ligation of necks of bilateral maxillary first molars. The gingival color and shape of the rats in two groups were observed， and the gingival bleeding index （BI）， periodontal probing depth （PD） and tooth mobility （TM） were measured and recorded. Hematoxylin-eosin（HE） staining was used to observe the pathomorphology of periodontium of the rats in two groups； micro-computed tomography （Micro-CT） was used to evaluate the alveolar bone resorption of the rats in two groups. HE and PAS staining were used to observe the pathomorphology of kidney tissue of the rats in two groups. Biochemical kits were used to assess kidney function indicators and detect the oxidative stress levels of the rats in two groups； Western blotting method was used to detect the expression levels of tumor necrosis factor-α （TNF-α） and nuclear factor kappa B （NF-κB） p65 protein in kidney tissue of the rats in two groups； real-time fluorescence quantitative PCR（RT-qPCR） and immunohistochemical method were used to detect the expression levels of SIRT1 and peroxisome-proliferator-activated receptor γ coactivator 1α （PGC-1α） mRNA and proteins in kidney tissue of the rats in two groups. Results The clinical indicators of peridontium tissue detection results showed that compared with control group， swollen gums， soft texture， deep periodontal pocket， bleeding gums and loosening of tooth of the rats in periodontitis group were found.The HE staining and Micro-CT results of peridontium tissue showed that compared with control group， attachment loss and alveolar bone absorption of the rats occurred in periodontitis group.The HE and PAS staining results of kidney tissue showed that compared with control group， the renal cystic space was dilated， the basement membrane of the glomerular was slightly thickened， the epithelial cells of the renal tubules were exfoliated， and the brush edge was destroyed in periodontitis group.The biochemical detection results showed that there was no significant difference in renal function indexes of the rats in two groups （P>0.05）； compared with control group，the levels of total antioxidative status （TAS），managanse superoxide dismutase （Mn-SOD） and glutathione （GSH） in kidney tissue of the rats in periodontitis group were decreased（P<0.01）， while the level of malondialdehyde （MDA） was increased （P<0.01）.The Western blotting results showed that compared with control group， the expression levels of TNF-α and NF-κB p65 proteins in kidney tissue of the rats in periodontitis group were increased （P<0.05 or P<0.01）.The RT-qPCR and immunohistochemistry results showed that compared with control group， the expression levels of SIRT1 and PGC-1α mRNA and proteins in kidney tissue of the rats in periodontitis group were decreased （P<0.05 or P<0.01）. Conclusion In the rat model of chronic periodontitis， the expression of SIRT1 in kidney tissue is down-regulated and the level of oxidative stress is increased， which aggravats the kidney injury of chronic periodontitis rats.

Key words: Chronic periodontitis, Kidney injury, Sirtuin 1, Rats

中图分类号:

R781.42

李鑫,丁旭,刘笑梦,刘歆婵,武洲,于维先. 沉默信息调节因子1对慢性牙周炎模型大鼠肾损伤的影响[J]. 吉林大学学报(医学版), 2022, 48(5): 1200-1208.

Xin LI,Xu DING,Xiaomeng LIU,Xinchan LIU,Zhou WU,Weixian YU. Effect of silencing information regulator 1 on kidney injury in chronic periodontitis model rats[J]. Journal of Jilin University(Medicine Edition), 2022, 48(5): 1200-1208.

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参考文献 27

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	LI L， ZHANG Y L， LIU X Y， et al. Periodontitis exacerbates and promotes the progression of chronic kidney disease through oral flora， cytokines， and oxidative stress［J］. Front Microbiol， 2021， 12： 656372.
3	HICKEY N A， SHALAMANOVA L， WHITEHEAD K A， et al. Exploring the putative interactions between chronic kidney disease and chronic periodontitis［J］. Crit Rev Microbiol， 2020， 46（1）： 61-77.
4	SHARMA P， FENTON A， DIAS I H K， et al. Oxidative stress links periodontal inflammation and renal function［J］. J Clin Periodontol， 2021， 48（3）： 357-367.
5	HIRANO K， SHIMBO T， KOMATSU Y， et al. Frequency of tooth brushing as a predictive factor for future kidney function decline［J］. J Nephrol， 2022， 35（1）： 191-199.
6	MIYATA Y， OBATA Y， MOCHIZUKI Y， et al. Periodontal disease in patients receiving dialysis［J］. Int J Mol Sci， 2019， 20（15）： E3805.
7	DESCHAMPS-LENHARDT S， MARTIN-CABEZAS R， HANNEDOUCHE T， et al. Association between periodontitis and chronic kidney disease： systematic review and meta-analysis［J］. Oral Dis， 2019， 25（2）： 385-402.
8	LI L， LIU X X， LI S S， et al. Tetrahydrocurcumin protects against sepsis-induced acute kidney injury via the SIRT1 pathway［J］. Ren Fail， 2021， 43（1）： 1028-1040.
9	周丰，陈野，陈晨，等. 沉默信息调节因子1调控牙周炎发生发展的机制［J］. 国际口腔医学杂志， 2021， 48（3）： 341-346.
10	TAMAKI N， CRISTINA ORIHUELA-CAMPOS R， INAGAKI Y， et al. Resveratrol improves oxidative stress and prevents the progression of periodontitis via the activation of the Sirt1/AMPK and the Nrf2/antioxidant defense pathways in a rat periodontitis model［J］. Free Radic Biol Med， 2014， 75： 222-229.
11	DU L， QIAN X， LI Y， et al. Sirt1 inhibits renal tubular cell epithelial-mesenchymal transition through YY1 deacetylation in diabetic nephropathy［J］. Acta Pharmacol Sin， 2021， 42（2）： 242-251.
12	KÖSE O， ARABACı T， YEMENOGLU H， et al. Influence of experimental periodontitis on cardiac oxidative stress in rats： a biochemical and histomorphometric study［J］. J Periodontal Res， 2017， 52（3）： 603-608.
13	YUE Y Y， LIU X C， LI Y， et al. The role of TLR4/MyD88/NF-κB pathway in periodontitis-induced liver inflammation of rats［J］. Oral Dis， 2021， 27（4）： 1012-1021.
14	HE W H， YOU M L， WAN W T， et al. Point-of-care periodontitis testing： biomarkers， current technologies， and perspectives［J］. Trends Biotechnol， 2018， 36（11）： 1127-1144.
15	向群，古丽努尔·阿吾提，李泽慧. MFG-E8介导的骨免疫在牙周炎中的作用研究进展［J］.解放军医学杂志，2021，46（6）：623-627.
16	CHOPRA A， SIVARAMAN K. An update on possible pathogenic mechanisms of periodontal pathogens on renal dysfunction［J］. Crit Rev Microbiol， 2019， 45（5/6）： 514-538.
17	KOSE O， KURT BAYRAKDAR S， UNVER B， et al. Melatonin improves periodontitis-induced kidney damage by decreasing inflammatory stress and apoptosis in rats［J］. J Periodontol， 2021， 92（6）： 22-34.
18	GALENO J G， FRANÇA L F C， SILVA F R PDA， et al. Renal alterations caused by ligature-induced periodontitis persist after ligature removal in rats［J］. J Periodontal Res， 2021， 56（2）： 306-313.
19	FRANÇA L F C， VASCONCELOS A C C G， SILVA F R PDA， et al. Periodontitis changes renal structures by oxidative stress and lipid peroxidation［J］. J Clin Periodontol， 2017， 44（6）： 568-576.
20	唐宋，张晓南. 牙周组织低氧环境与牙周炎发生发展的研究进展［J］. 同济大学学报（医学版），2021，42（2）：285-290.
21	SCZEPANIK F S C， GROSSI M L， CASATI M，et al. Periodontitis is an inflammatory disease of oxidative stress： we should treat it that way［J］. Periodontol 2000， 2020， 84（1）： 45-68.
22	SILVA J CDA， MUNIZ F W M G， OBALLE H J R， et al. The effect of periodontal therapy on oxidative stress biomarkers： a systematic review［J］. J Clin Periodontol， 2018， 45（10）： 1222-1237.
23	ERDEMLI Z， ERDEMLI M E， GUL M， et al. Ameliorative effects of crocin on the inflammation and oxidative stress-induced kidney damages by experimental periodontitis in rat［J］. Iran J Basic Med Sci， 2021， 24（6）： 825-832.
24	TIRICHEN H， YAIGOUB H， XU W W， et al. Mitochondrial reactive oxygen species and their contribution in chronic kidney disease progression through oxidative stress［J］. Front Physiol， 2021， 12： 627837.
25	程志芬，杨磊，郭留云. 慢性牙周炎患者牙龈组织中Shh的表达及意义［J］.郑州大学学报（医学版），2021，56（5）：614-618.
26	VO T T T， CHU P M， TUAN V P， et al. The promising role of antioxidant phytochemicals in the prevention and treatment of periodontal disease via the inhibition of oxidative stress pathways： updated insights［J］. Antioxidants （Basel）， 2020， 9（12）： E1211.
27	PANES J D， GODOY P A， SILVA-GRECCHI T，et al.Changes in PGC-1α/SIRT1 signaling impact on mitochondrial homeostasis in amyloid-beta peptide toxicity model［J］. Front Pharmacol， 2020， 11： 709.

Group	PD（l/mm）	BI	TM/degree
		0 1 2 3 4 5	<Ⅰ Ⅰ Ⅱ Ⅲ
Control	0.62±0.07	10 0 0 0 0 0	10 0 0 0
Periodontitis	2.13±0.24 ^*	1 2 3 2 1 1	1 5 3 1

Group	SIRT1	PGC-1α
Control	1.01±0.09	1.00±0.06
Periodontitis	0.50±0.02 ^**	0.76±0.03 ^*

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