發(fā)布時間：2018-01-09 04:15

  本文關鍵詞：不同牙周狀態(tài)下牙齦組織內(nèi)質(zhì)網(wǎng)應激水平的差異性探究　出處：《第四軍醫(yī)大學》2016年碩士論文　論文類型：學位論文

  更多相關文章： 內(nèi)質(zhì)網(wǎng)應激 慢性牙齦炎 慢性牙周炎 侵襲性牙周炎

【摘要】：大量研究表明細胞凋亡參與了牙周疾病的病理生理過程,而與細胞凋亡相關的通路主要分為線粒體途徑、死亡受體途徑、顆粒酶B途徑以及近年來新發(fā)現(xiàn)的內(nèi)質(zhì)網(wǎng)應激(endoplasmic reticulum stress,ERS)途徑。ERS是組織及細胞在不利的生存環(huán)境下產(chǎn)生的一種應激性反應,其目的在于維持細胞穩(wěn)態(tài),主要途徑為保護性的非折疊蛋白反應(unfolded protein response,UPR)。UPR可以通過蛋白質(zhì)生成減慢、折疊功能加強、未折疊蛋白降解等途徑適應內(nèi)質(zhì)網(wǎng)應激,對細胞起保護作用,但是當刺激持續(xù)存在或強度過大,超過細胞自我調(diào)節(jié)能力時則會誘導細胞發(fā)生凋亡,細胞整體由促生存向促凋亡方向發(fā)生轉(zhuǎn)變。牙齦作為唯一暴露于口腔及各類牙周疾病起始部位的牙周組織,探究牙齦組織從健康到慢性牙齦炎再到慢性牙周炎及侵襲性牙周炎的發(fā)展過程中其內(nèi)質(zhì)網(wǎng)應激水平發(fā)生的改變具有重要的臨床意義。當牙周疾病發(fā)生時,牙周致病菌的致病成分及其產(chǎn)生的毒素等可以使牙齦上皮的多種粘結蛋白如上皮鈣黏素和封閉蛋白-1等的表達水平出現(xiàn)異常,上皮細胞間的相互連接受到破壞發(fā)生,牙齦結合上皮通透性顯著增加,導致菌斑微生物更易于穿過牙齦上皮的物理屏障,到達上皮深層組織,從而促進牙周炎的發(fā)展;此外,牙齦上皮分泌的多種免疫因子如抗菌肽LL-37、人β防御素等在抵御菌斑微生物的入侵,預防牙周病的發(fā)展過程中均起到了重要作用。目前已有文獻報道ers參與了慢性牙周炎的發(fā)病過程,有研究報道指出在慢性牙周炎情況下內(nèi)質(zhì)網(wǎng)應激上游標志物grp78及下游凋亡相關因子chop升高,但是健康與慢性牙齦炎,侵襲性牙周炎情況下ers水平的差異表達情況尚無相關報道。目的初步探討健康、慢性牙齦炎、慢性牙周炎及侵襲性牙周炎牙齦組織中內(nèi)質(zhì)網(wǎng)應激水平是否存在差異。方法探究不同狀態(tài)下牙齦組織中內(nèi)質(zhì)網(wǎng)應激標志物grp78、perk、atf4、chop的表達變化:臨床收集健康、慢性牙齦炎、慢性牙周炎及侵襲性牙周炎患者的牙齦,通過免疫組化染色對牙齦組織中內(nèi)質(zhì)網(wǎng)應激標志物的表達水平進行定性分析;實時定量pcr檢測牙齦組織中grp78、perk、atf4、chop的mrna表達量;蛋白質(zhì)印跡法檢測牙齦組織中grp78、perk、atf4、chop的蛋白表達量。結果1.免疫組化結果顯示:grp78及chop陽性表達細胞率均為侵襲性牙周炎組慢性牙周炎組慢性牙齦炎組牙周健康組,差異具有統(tǒng)計學意義(p0.05);perk及atf4牙齦組織中陽性細胞表達率的統(tǒng)計結果為慢性牙周炎慢性牙齦炎健康及侵襲性牙周炎組,且差異有統(tǒng)計學意義(p0.05),健康組及侵襲性牙周炎組中perk及atf4兩種因子表達量的差異沒有統(tǒng)計學意義(p0.05)。2.real-timepcr結果顯示:grp78、chopmrna表達量呈現(xiàn)一致性,均表現(xiàn)為健康組慢性牙齦炎組慢性牙周炎組侵襲性牙周炎組,且差異均有統(tǒng)計學意義(p0.05)。perkmrna表達量為健康組與侵襲性牙周炎組間無顯著性差異(p0.05),慢性牙齦炎組及慢性牙周炎組表達均有升高,且慢性牙周炎組高于慢性牙齦炎組,差異均有統(tǒng)計學意義(p0.05)。atf-4mrna的表達結果為健康組及侵襲性牙周炎組間表達量未見顯著性差異(p0.05);慢性牙齦炎組與慢性牙周炎組高于健康及侵襲性牙周炎組,差異有統(tǒng)計學意義(P0.05);但慢性牙齦炎組及慢性牙周炎組間表達未見顯著性差異(P0.05)。3.蛋白印跡法結果顯示:GRP78、CHOP蛋白在健康組、慢性牙齦炎組、慢性牙周炎組及侵襲性牙周炎組的表達量為:健康組慢性牙齦炎組慢性牙周炎組侵襲性牙周炎組,且差異有統(tǒng)計學意義(P0.05);PERK、ATF-4蛋白的表達情況為:健康組與侵襲性牙周炎組慢性牙齦炎組慢性牙周炎組,組間差異有統(tǒng)計學意義(P0.05),而健康組及侵襲性牙周炎兩組間PERK、ATF-4蛋白的表達量的差異沒有統(tǒng)計學意義(P0.05)。結論1、不同狀態(tài)下牙齦組織中的內(nèi)質(zhì)網(wǎng)應激水平存在差異,且表現(xiàn)為健康組慢性牙齦炎組慢性牙周炎組侵襲性牙周炎。2、內(nèi)質(zhì)網(wǎng)應激PERK通路的水平為健康組、侵襲性牙周炎組慢性牙齦炎組慢性牙周炎組。3、侵襲性牙周炎患者牙齦的內(nèi)質(zhì)網(wǎng)應激水平高于慢性牙周炎,但兩者之間引起內(nèi)質(zhì)網(wǎng)應激反應的通路可能有所不同。
[Abstract]:A large number of studies show that apoptosis is involved in the pathogenesis of periodontal disease, and apoptosis related pathways are mainly divided into the mitochondrial pathway, death receptor pathway, Granzyme B pathway and recently discovered endoplasmic reticulum stress (endoplasmic reticulum stress, ERS).ERS is a stress reaction of tissue and cell in the adverse environment, its purpose is to maintain the cell homeostasis, the main way for the protection of the unfolded protein response (unfolded protein response, UPR.UPR) can be generated by slow protein folding function strengthen, unfolded protein degradation pathway to endoplasmic reticulum stress, protective effect on cells, but when the stimulus persist or intensity is too large, more than cell self-regulation would induce cell apoptosis, cell apoptosis by promoting the survival to the overall direction of occurrence of gum change. As the only exposure to the oral cavity and all kinds of periodontal disease initiation site of periodontal tissue, explore the gingival tissue from healthy to chronic gingivitis and chronic periodontitis development and aggressive periodontitis in the endoplasmic reticulum stress level changes has important clinical significance. When periodontal disease occurs, the pathogenic composition of periodontal pathogens and the toxin can make a variety of adhesive protein such as gingival epithelial expression of E-cadherin and -1 protein closed abnormal epithelial cells, the inter connection damage occurs, the junctional epithelial permeability increased significantly, resulting in plaque microorganisms more easily through the physical barrier of gingival epithelium, epithelial to deep tissue thus, to promote the development of periodontitis; in addition, a variety of immune factors such as gingival epithelial secretion of antimicrobial peptide LL-37 of human beta defensin in resisting microbial plaque The invasion, development process of prevention of periodontal disease has played an important role. At present, it has been reported that ers is involved in the pathogenesis of chronic periodontitis, studies have reported that in the case of chronic periodontitis upstream of endoplasmic reticulum stress related factors of GRP78 signs and downstream apoptosis of chop increased, but the health and chronic gingivitis, there is no related situation reported the differential expression of ers level of aggressive periodontitis cases. Objective to explore the health, chronic gingivitis, the existence of ER stress levels in chronic periodontitis and aggressive periodontitis gingival tissue. Methods on gingival tissue differences under different states of endoplasmic reticulum stress markers GRP78, perk, ATF4, chop expression changes: collection of clinical health, chronic gingivitis and chronic periodontitis and aggressive periodontitis gingival, by immunohistochemical staining of the endoplasmic reticulum stress markers in gingival tissue The qualitative analysis of the expression level; GRP78, real-time quantitative PCR detection of perk ATF4 in gingival tissue, the expression of chop, mRNA; GRP78, Western blotting detection of gingival tissue in perk, ATF4, chop protein expression was 1.. The results of immunohistochemistry showed that GRP78 and chop positive cells were invasive periodontitis group of chronic periodontitis group of chronic gingivitis group periodontal health group, the difference was statistically significant (P0.05); perk and ATF4 positive cells in gingival tissue expression rate of statistical results for chronic gingivitis and chronic periodontitis and healthy aggressive periodontitis group, and the difference was statistically significant (P0.05), perk and ATF4 in health group and invasive in periodontitis group two factor expression difference was not statistically significant (P0.05.2.real-timepcr) the results showed that GRP78, chopmrna expression showed consistency, showed a healthy group of chronic gingivitis group of chronic periodontal The inflammation group of aggressive periodontitis group, and the differences were statistically significant (P0.05) the expression level of.Perkmrna was no significant health group and aggressive periodontitis group differences (P0.05), chronic gingivitis and chronic periodontitis group group expression were increased, and the chronic periodontitis group was higher than that of chronic gingivitis group, there were statistically significant differences (P0.05) results.Atf-4mrna the expression is no significant difference between healthy group and aggressive periodontitis group (P0.05 group); chronic gingivitis and chronic periodontitis group was higher than that of healthy and aggressive periodontitis group, the difference was statistically significant (P0.05); but the chronic gingivitis group and chronic periodontitis group was no significant difference between the expression of.3. protein (P0.05) Western blot results showed that GRP78, CHOP protein in healthy group, chronic gingivitis group, chronic periodontitis group and the expression of aggressive periodontitis group were: group of chronic gingivitis group of chronic periodontal health Periodontitis group of aggressive periodontitis group, and the difference was statistically significant (PERK, P0.05); the expression of ATF-4 protein: healthy group and aggressive periodontitis group of chronic gingivitis group of chronic periodontitis group, there was significant difference between the groups (P0.05), and the healthy group and aggressive periodontitis PERK between the two groups, the difference expression the expression of ATF-4 was not statistically significant (P0.05). Conclusion: 1, endoplasmic reticulum stress level in gingival tissue under different conditions in the existence of differences, and performance for the healthy group of chronic gingivitis group of chronic periodontitis group of aggressive periodontitis.2, PERK pathway of endoplasmic reticulum stress levels as healthy group, aggressive periodontitis group of chronic gingivitis and chronic periodontitis group.3, endoplasmic reticulum stress levels in patients with aggressive periodontitis gingival than chronic periodontitis, but between the two pathways caused by endoplasmic reticulum stress response may be different.

【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R781.4

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