Levels of Hsp60 in periodontal tissue of rats: influence of injections of hyaluronic acid

O. V. Kopchak, L. Yakovenko, N. Marchenko, I.V. Кovach, E. Pavlenko, O.A. Nimenko, I. Kroupskaya, V. Filonenko
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Abstract

Heat shock protein 60 (Hsp60) is considered as one of the possible autoantigens involved in the pathogenesis of a number of chronic diseases including periodontal diseases. The application of hyaluronic acid or hyaluronan (HA) in the treatment of periodontitis has been evaluated in several clinical trials, however, the effect of hyaluronic acid on heat shock protein 60 level in periodontal soft tissues has not been studied. The aim of this work was to evaluate the effect of HA injections on levels of Hsp60 in periodontal tissue of the rats. Samples of periodontal tissue of mandibular incisors of adult male Wistar rats at 10-12 months of age were investigated. Rats were distributed into the control group and the periodontitis group. Visual manifestations of hyperemia of the gums around the incisors were the criterion for selecting animals into the periodontitis group. There were two subgroups in the control group: intact rats (I); intact rats after HA “hyaDENT BG” 1.0 MDa (BioScience GmbH, Germany) treatment (I+“G-1.0”). There were four subgroups in the periodontitis group: rats with periodontitis (P); rats with periodontitis after HA “hyaDENT BG” 1.0 MDa (BioScience GmbH, Germany) treatment (P+“G-1.0”); rats with periodontitis after HA “SERTOBEC” 2.4 MDa (S.C. Rompharm Company S.R.L., Romania) treatment (P+“S-2.4”); rats with periodontitis after HA “SERTOBEC Tendon” 2.4 МDа (S.C. Rompharm Company S.R.L., Romania) treatment (P+“ST-2.4”). There were three animals in each subgroup. Rats were injected 0.05 ml HA in the area of alveolar processus of central incisors once a week, three times. Levels of Hsp60 in total lysates of periodontal tissue were tested by Western blotting method before and after the treatment with HA (one month after the last injection). There was no significant difference between levels of Hsp60 in total lysates of periodontal tissue of intact rats and rats with periodontitis before treatment (p>0.05). Rats with periodontitis showed decreased inflammation in the periodontal tissue after treatment with HA with different molecular weight. Intact rats and rats with periodontitis which were treated with HA “hyaDENT BG” 1.0 MDa showed reduced levels of Hsp60 in total lysates of periodontal tissue comparatively with levels of Hsp60 before treatment (by 15.4 and 10.7 times respectively, p<0.001). Rats with periodontitis which were treated with HA “SERTOBEC” 2.4 MDa or HA “SERTOBEC Tendon” 2.4 MDa also showed reduced levels of Hsp60 in total lysates of periodontal tissue com­paratively with levels of Hsp60 before treatment (by 21.3 and 16.4 times respectively, p<0.001). Rats with pe­riodontitis showed the decrease in inflammation in periodontal tissue after treatment with HA with different molecular weight. Injections of HA has contributed to reduce levels of Hsp60 in periodontal tissue of intact rats and rats with periodontitis.
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注射透明质酸对大鼠牙周组织Hsp60水平的影响
热休克蛋白60 (Hsp60)被认为是参与包括牙周病在内的许多慢性疾病发病机制的可能的自身抗原之一。透明质酸或透明质酸(HA)在牙周炎治疗中的应用已经在一些临床试验中得到了评价,然而,透明质酸对牙周软组织热休克蛋白60水平的影响还没有研究。本研究的目的是评价透明质酸注射对大鼠牙周组织Hsp60水平的影响。对10 ~ 12月龄成年雄性Wistar大鼠下颚门牙牙周组织进行了研究。将大鼠分为对照组和牙周炎组。门牙周围牙龈充血的视觉表现是选择牙周炎组动物的标准。对照组分为两个亚组:完整大鼠(I);HA“hyaDENT BG”1.0 MDa (BioScience GmbH, Germany)处理后(I+“G-1.0”)。牙周炎组分为4个亚组:牙周炎大鼠(P);HA“hyaDENT BG”1.0 MDa (BioScience GmbH, Germany)治疗后的牙周炎大鼠(P+“G-1.0”);HA“SERTOBEC”2.4 MDa (S.C. Rompharm Company S.R.L, Romania)治疗后的牙周炎大鼠(P+“S-2.4”);HA“SERTOBEC肌腱”2.4 МDа (S.C. Rompharm Company S.R.L, Romania)治疗后的牙周炎大鼠(P+“ST-2.4”)。每个亚组有三只动物。大鼠中切牙牙槽突区注射HA 0.05 ml,每周1次,共3次。采用Western blotting法检测HA治疗前后(末次注射后1个月)牙周组织总裂解液中Hsp60水平。治疗前牙周炎大鼠与完整大鼠牙周组织总裂解物中Hsp60水平差异无统计学意义(p>0.05)。不同分子量透明质酸对牙周炎大鼠牙周组织的炎症反应均有所减轻。与治疗前相比,HA“hyaDENT BG”1.0 MDa治疗的正常大鼠和牙周炎大鼠牙周组织总裂解物中Hsp60水平降低(分别降低15.4倍和10.7倍,p<0.001)。与治疗前相比,HA“SERTOBEC”2.4 MDa或HA“SERTOBEC肌腱”2.4 MDa治疗的牙周炎大鼠牙周组织总溶解物中Hsp60水平也降低(分别降低21.3倍和16.4倍,p<0.001)。牙周炎大鼠经不同分子量透明质酸处理后,牙周组织炎症反应明显减轻。注射透明质酸有助于降低完整大鼠和牙周炎大鼠牙周组织中Hsp60的水平。
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