| Fusobacterium nucleatum that has been found in the oral cavity,spleen,brain,and lungs of body abscesses,is a common Gram-negative anaerobic bacterium.It has a strong film-forming ability to form biofilm through copolymerization,which aggravates the difficulty of disease cure.F.nucleatum has been found to damage periodontal and gingival tissues,causing oral diseases including bad breath and periodontitis.In recent years,it has been found that F.nucleatum is closely related to the occurrence and development of colorectal cancer and can also affect the intestinal flora.Egg yolk immunoglobulin(Ig Y)is isolated from egg yolk that hens lay.Hens are injected a specific antigen or specific pathogens to stimulate the production of antibodies that transfer to egg yolk.Ig Y has structural similarity with mammalian Ig G and highly efficient immune properties.Due to its stability,cheap and accessibility,Ig Y have been applied in various fields.Based on its immunological characteristics,this study used anti-F.nucleatum Ig Y to intervene related halitosis,periodontitis and colorectal cancer induced by F.nucleatum to verify whether there is a good therapeutic effect and to explain its brief related mechanisms.Ig Y inhibits the proliferation of F.nucleatum.F.nucleatum is inactivated and is added auxiliary reagents to make a vaccine for injection into hens,collecting egg yolks,centrifuging to remove fat,removing residual fat according to 1: 6 water,precipitating with 50% saturated ammonium sulfate,and then precipitating with 14% sodium sulfate.Finally,Ig Y was obtained by dialysis through a 100 k Da ultrafiltration tube and verified by SDS-PAGE and native-PAGE.The molecular weight of natural Ig Y was about 180 k Da,and the heavy chain was about 70 k Da and the light chain was about 25 k Da.F.nucleatum was cultured and identified by Gram staining.The growth curve was measured,which ensured the follow-up experiments.Ig Y titer was as high as 25600 by ELISA,which had a good inhibitory effect on the growth of F.nucleatum.Ig Y treated the halitosis induced by F.nucleatum.Based on in vitro and animal models,halitosis models were induced using F.nucleatum,the major responsibility for halitosis.In the in vitro simulation system,the artificial saliva system that was verified the stability and effectiveness was used.After induction with F.nucleatum,Ig Y can effetely inhibit the growth of F.nucleatum in artificial saliva and reduce VSCs produced by F.nucleatum in artificial saliva.The changes of fatty acid in the bacterial fluid were detected by GC / MS,it is speculated that Ig Y affects the metabolism of F.nucleatum fatty acids and can treat bad breath.Using F.nucleatum to coat periodontal gums and adding physical methods for one month in the halitosis rat model.A good bad breath model was obtained through sensory evaluation.After Ig Y treatment,the halitosis index significantly decreased.Subsequently,SPME-GC/MS was used to detect the changes of volatile organic compounds in the saliva of animal models.The peak areas of were Qualitative and semi-quantitative calculations based on the NIST database.4 alcohols,4 carboxylic acids,3 ketones,and 2 Aldehydes,2 esters,and 1 nitrogen-containing compound are basically the same as described in the previous literature.After the Ig Y intervention,although the ketones and aldehydes did not change,the content of acetic acid,propionic acid and hexanoic acid was significantly reduced.Another major odorant(ammonia)was significant remission after treatment with high doses(40 mg / m L)and low doses.(10mg / m L).Ig Y remedied the periodontitis induced by F.nucleatum.After the rat periodontitis model was treated with Ig Y,X-rays showed that the gaps between the teeth could be significantly reduced;the HE staining shows that alveolar bone resorption was greatly improved,periodontal ligament tissue inflammation decreased and collagen fibers became orderly,which indicated that Ig Y has a better treatment for F.nucleatuminduced periodontitis.Ig Y acts on F.nucleatum to reduce the invasion and stimulation of the body and reduces the release of inflammatory factors like IL-6 and TNF-?.Ig Y relieves colorectal cancer with F.nucleatum intervention.To study the effects of Ig Y on mouse CRC,we used intraperitoneal injection of azomethane,drinking dextran sodium sulfate solution to promote enteritis,and induction of F.nucleatum to construct a mouse colorectal cancer model.The number of colorectal tumors and tumor cell proliferation of mice were significantly increased than control group,and the close relationship between inflammation and colorectal cancer was also demonstrated.In this experiment,it was found that the relative expression levels of TNF-?,COX-2 and IL-8 in colorectal tissue and the blood content in colorectal cancer serum induced by F.nucleatum were significantly higher than those control group.Rat weight,tumor number,hair color,and other symptoms were significantly different from the control group.But after Ig Y intervention,the body weight recovered better than before,the number of tumors decreased,and the TNF-?,COX-2,and IL-8were decreased.The decrease in levels indicates that F.nucleatum has an important effect on inflammation,and Ig Y can alleviate the inflammation in colorectal tissues through the NF-?B pathway to reduce the risk of colorectal tumors. |
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