牙髓炎模型大鼠血清褪黑素水平及其对牙髓炎症反应的抑制作用

doi:10.13481/j.1671-587x.20190624

摘要/Abstract

摘要： 目的：探讨牙髓炎大鼠血清褪黑素水平对核苷酸结合寡聚化结构域样受体蛋白3/含半胱氨酸的天冬氨酸蛋白水解酶1（NLRP3/caspase-1）信号通路的影响，阐明褪黑素对牙髓炎的作用机制。方法：将130只大鼠随机分为对照组（n=40）、牙髓炎组（n=60）和牙髓炎+褪黑素组（n=30），其中30只牙髓炎大鼠随机分为牙髓炎1 d组、牙髓炎3 d组和牙髓炎5 d组，采用开髓暴露法建立牙髓炎大鼠模型，牙髓炎+褪黑素组大鼠开髓术后腹腔注射褪黑素。HE染色确定各组大鼠牙髓炎建模情况，ELISA法检测各组大鼠血清白细胞介素1β（IL-1β）和褪黑素水平以及牙髓组织中IL-1β水平，RT-PCR法检测各组大鼠牙髓组织中NLRP3和caspase-1 mRNA表达水平，Western blotting法检测各组大鼠牙髓组织中NLRP3和caspase-1蛋白表达水平。结果：HE染色，对照组大鼠牙髓组织正常，牙髓炎组大鼠牙髓有脓腔形成，可见大量炎性细胞聚集。牙髓炎1 d和3 d组大鼠血清IL-1β水平明显高于对照组（P<0.05），褪黑素水平明显低于对照组（P<0.05）；牙髓炎3 d组大鼠血清IL-1β水平明显低于牙髓炎1 d组（P<0.05），褪黑素水平明显高于牙髓炎1 d组（P<0.05）；牙髓炎组和牙髓炎+褪黑素组大鼠牙髓组织中IL-1β水平明显高于对照组（P<0.05），且牙髓炎+褪黑素组大鼠牙髓组织中IL-1β水平明显低于牙髓炎组（P<0.05）；牙髓炎组和牙髓炎+褪黑素组大鼠牙髓组织中NLRP3和caspase-1mRNA和蛋白表达水平均明显高于对照组（P<0.05），且牙髓炎+褪黑素组大鼠牙髓组织中NLRP3和caspase-1mRNA和蛋白水平明显低于牙髓炎组（P<0.05）。结论：牙髓炎大鼠血清褪黑素水平降低，褪黑素可通过抑制NLRP3/caspase-1信号通路降低牙髓炎大鼠牙髓组织的炎症反应。

关键词: 牙髓炎, 褪黑素, 核苷酸结合寡聚化结构域样受体蛋白3, 含半胱氨酸的天冬氨酸蛋白水解酶1, 白细胞介素1β

Abstract: Objective: To investigate the effect of serum melatonin level in the rats with pulpitis on the nucleotide-binding oligomerization domain-like receptor protein 3/cysteine-containing aspartate proteolytic enzyme 1 (NLRP3/caspase-1) signaling pathway,and to explore the mechanism of the effect of melatonin on pulpitis. Methods: A total of 130 rats were divided into control group (n=40),pulpitis group (n=60) and pulpitis+melatonin group (n=30).Thirty rats with pulpitis were randomly divided into pulpitis 1 d group,pulpitis 3 d group and pulpitis 5 d group.The pulpitis rat model was established by pulpotomy exposure method, and the rats in pulpitis+melatonin group were intraperitoneally injected with melatonin after intramedullary surgery.HE staining was used to confirm the modeling of pulpitis;the serum levels of interleukin-1β(IL-1β) and melatonin and the levels of IL-1β in dental pulp tissue of the rats in various groups were detected by ELISA method, the expresion levels of NLRP3 and caspase-1 mRNA in dental pulp tissue of the rats in various groups were detected by RT-PCR method,and the expression levels of NLRP3 and caspase-1 proteins in dental pulp tissue of the rats in various groups were detected by Western blotting method. Results: The HE staining results showed that in control group,the dental pulp tissue of the rats was normal;in pulpitis group,the cavities in the dental pulp and a large number of inflammatory cells were found.The serum IL-1β levels of the rats in pulpitis 1 d group and pulpitis 3 d group were obviously higher than those in control group (P<0.05),and the serum melatonin levels were obviously lower than those in control group (P<0.05);the serum IL-1β level of the rats in pulpitis 3 d group was obviously lower than that in pulpitis 1 d group (P<0.05),and the serum melatonin level was obviously higher than that in pulpitis 1 d group (P<0.05).The levels of IL-1β in dental pulp tissue of the rats in pulpitis group and pulpitis+melatonin group were obviously higher than those in control group (P<0.05),the level of IL-1β in dental pulp tissue of the rats in pulpitis+melatonin group was obviously lower than that in pulpitis group (P<0.05). The expression levels of NLRP3 and caspase-1 mRNA and proteins in dental pulp tissue of the rats in pulpitis group and pulpitis+ melatonin group were obviously higher than those in control group (P<0.05),and the levels of NLRP3 and caspase-1 mRNA and proteins in dental pulp tissue of the rats in pulpitis+melatonin group were obviously lower than those in pulpitis group (P<0.05). Conclusion: The serum melatonin level in the rats with pulpitis is reduced,and melatonin can reduce the inflammatory response in dental pulp tissue of the rats with pulpitis by inhibiting NLRP3/caspase-1 signaling pathway.

Key words: pulpitis, melatonin, nucleotide-binding oligomerization domain-like receptor proein 3, cysteine-containing aspartate proteolytic enzyme 1, interleukin-1β

中图分类号:

R781.3

韩耀伦, 王璐, 马欣. 牙髓炎模型大鼠血清褪黑素水平及其对牙髓炎症反应的抑制作用[J]. 吉林大学学报(医学版), 2019, 45(06): 1334-1339.

HAN Yaolun, WANG Lu, MA Xin. Levels of serum melatonin in rats with pulpitis and its inhibitory effect on dental inflammation[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1334-1339.

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