Study On The Anti-Inflammation And Antioxidation Mechanism Of Coenzyme Q10 Extracted From Yunnan Tobacco Leaf

Objective:The purpose of the study is to investigate the effect and mechanism of coenzyme Q10 (CoQ10), extracted from Yunnan tobacco leaf,on the periodontitis through inflammation model in vitro and vivo.It would provide theoretical basis for the prevention and treatment of periodontitis.Methods:①126 Specific-pathogen-free male rats were used for experiments at the age of 6-8 weeks and were divided into seven groups of 18 rats each. One group was left untreated and six groups,contained Model group (P), Tinidazole group (T) and Vitamin C group (VC), CoQ10 low dose group (LQ), CoQ10 medium dose group (MQ), CoQ10 high dose group (HQ),had their bilateral maxillary second molar tooth ligated and sticky sugar diet fed. LQ, MQ, HQ intragastric adiministration with 15,30,60 mg/kg of CoQ10 solution respectively; T intragastric adiministration with 200 mg/kg on the first day, the next 100 mg/kg of Tinidazole solutio; VC intragastric adiministration 100 mg/kg Vitamin C solution; N and P intragastric adiministration with the same amount of oil.Then each group were randomly selected 6 rats and sacrificed at three different time points post-intragastric adiministration (2w,4w and 6w).Blood sample was collected through the abdominal aorta and centrifugal to get plasma for detecting three inflammatory cytokines:interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-a), prostaglandin E2(PGE2) and three antioxidant indices:superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA).Maxilla samples were collected to detect alveolar bone loss value and perform hematoylin-eosin(HE) staining and tartrate-resistant acid phosphatase(TRAP) staining to observe the inflammatory response and osteoclast count.②Escherichia coli lipopolysaccharide LPS stimulated macrophage RAW264.7 to construct inflammatory model in vitro.The experimental group was divided into model group;Coenzyme Q10 group with different concentrations(1.875,3.75,7.5,15, 30μmol/L); Vitamin C group(200μmol/L). model group and treatment group were incubated with LPS(1 mg/L) for 24h. After that, the cell culture supernatant was collected, ELISA method for the detection of inflammatory factor IL-1β、TNF-α and PGE2 content; collecting cells for the detection of antioxidant indexes such as SOD, GSH-Px, MDA. All data were statistically analyzed by SPSS 17.0 data statistics software.Results:1. After 4 weeks modeling, modeling rats’gingival Index(GI), alveolar bone loss(ABL) were higher than the N group(P< 0.05); HE staining showed that the separation between the epithelium and the tooth in rat model, inflammatory cell infiltration, elongation of epithelial nail to connective tissue, disorder of collagen fiber arrangement, alveolar crest damage absorption. The results represent the success of the model.2. During the observation period of 6 weeks, there was no significant difference about the ABL value and the number of osteoclasts between CoQ10 each dose group and P group (P>0.05). The ABL value and the number of osteoclasts of T group and VC group were similar to CoQ10 each dose group(P>0.05).3. High, medium and low doses of CoQ10 can significantly inhibit the level of IL-1β, TNF-a, PGE2 in peripheral blood of rats with periodontitis, improve the activity of SOD and GSH-Px and decrease the content of MDA (P<0.05). CoQ10, tinidazole and vitamin C have similar ability to inhibit three kinds of inflammatory factors(P> 0.05). The effect of CoQ10 group was similar to VC group (P>0.05).4. CoQ10 could significantly inhibit the secretion of EL-1β, TNF-α, PGE2 in RAW264.7 cells stimulated by LPS, and increase the activity of SOD and GSH-Px in the cells and decreased MDA content(P<0.05). And the effect is similar to that of vitamin C(P>0.05).Conclusions:1. By means of the ligation of the bilateral maxillary second molar teeth and the high sugar diet,experimental periodontitis model can be successfully established after 4 weeks.2. During the observation period of 6 weeks, the effect of CoQ10 on the loss of alveolar bone and the number of osteoclasts in the alveolar bone of rats was not found, It is still not considered that CoQ10 can improve the bone resorption caused by periodontitis.3. In this study, we found that CoQ10 can significantly reduce the level of IL-1β, TNF-α and PGE2, significantly increased SOD, GSH-Px activity, reduce the amount of MDA in plasma and improve the oxidative stress of the body, which made the inflammatory reaction decrease.4. CoQ 10 can inhibit the release of IL-1β、TNF-α and PGE2, increase the activity of SOD and GSH-Px and decrease the content of MDA in vitro. Therefore CoQ10 has the ability of anti inflammation and anti oxidation, which can inhibit oxidative stress and reduce the inflammatory reaction.