| Objective:The pathogenesis of chemotherapy or radiotherapy induced oral mucositis has not been fully clarified.The pathogenesis of the two is similar,which is described by a five stage pathogenesis model.And there is no effective conventional treatment based on evidence-based medicine.Previous studies have shown that "Kui-Yang-Gel" can effectively improve chemotherapy or radiotherapy-induced oral mucositis,but the mechanism is not yet clear.From the two aspects of clinical efficacy analysis and mechanism exploration,we studied the effect of "Kui-Yang-Gel" on chemotherapy or radiotherapy-induced oral mucositis,so as to provide reference for its clinical application.Methods:1 In clinical studies,based on published literature,Meta-analysis was used to evaluate the clinical efficacy of "Kui-Yang-Gel".In the database of CNKI,Wan Fang,VIP and PubMed,we searched the literature about "Kui-Yang-Gel" for treating radiotherapy-induced dermatitis and mucositis up to March 2022.After the completion of literature screening,methodological evaluation,meta-analysis and publication bias risk evaluation were carried out for the literature that met the criteria.2 In the comparative study of RIOM model,27 Wistar rats were randomly divided into Control group,18-Gy group and 30-Gy group.After radiation,the body weight,food intake and oral mucositis index(OMI)of rats were recorded and analyzed.The samples were taken in batches on the 7th,10th and 13th days after irradiation,and the tongue tissue was stained with H&E for pathological evaluation.3 In the comparative study of CTOM model,45 Wistar rats were randomly divided into 5 groups,including Control group,IP group,IP +MS group,IP+CS group and Ⅳ group.After the model established,the body weight,food intake and OMI of rats were recorded and analyzed.The oral mucosa was stained by H&E and the expression of TNF-α and IL-6 was detected by immunohistochemical staining.4 According to the RIOM model establishment method determined above,36 Wistar rats were randomly divided into 4 groups,including Control group,model group,KFX group and"Kui-Yang-Gel" group,in the study of "Kui-Yang-Gel" in the treatment of RIOM model rats.After model establishment,rats in each group were given corresponding drug intervention every day,and their body weight,food intake and OMI were recorded and analyzed,and saliva samples were collected.The samples were taken 7 days after irradiation.Tongue mucosa was taken for H&E staining,and oral flora was detected by 16S RNA sequencing technology,transcriptome was detected by transcriptional sequencing analysis,and the expression of related protein of TLR4-MyD88-TRAF6-NF-κB pathway was detected by Western blotting.The expression of IFN-γ,TNF-α,IL-1β,MCP-1,IL-13,IL-10,IL-1α,IL-2,IL-4 and IL-6 was detected by protein quantitative chip.5 According to the CTOM model establishment method determined above,36 Wistar rats were randomly divided into 4 groups,including Control group,model group,KFX group and"Kui-Yang-Gel" group,in the study of "Kui-Yang-Gel" in the treatment of CTOM model rats.After model establishment,rats in each group were given corresponding drug intervention every day,and their body weight,food intake and OMI were recorded and analyzed,and saliva samples were collected.The samples were taken on day 7 of the experiment.Buccal mucosa was taken for H&E staining,and oral flora was detected by 16S RNA sequencing technology,transcriptome was detected by transcriptional sequencing analysis,and the expression of related protein of Wnt2/β-catenin、NF-κB was detected by Western blotting.The expression of IFNγ,TNF-α,IL-1β,MCP-1,IL-13,IL-10,IL-1α,IL-2,IL-4 and IL-6 was detected by protein quantitative chip.Results:1 A total of 11 literatures in Chinese were included in the Meta-analysis.There are heterogeneity and risk of publication bias among the literatures.The results suggest that compared with conventional treatment,"Kui-Yang-Gel" can effectively reduce the incidence of moderate to severe radiodermatitis[OR=0.18,95%CI(0.08,0.38),P<0.00001];reduce the incidence of dermatitis before the cumulative radiation dose up to 40Gy,delay the occurrence of skin injury[OR=0.20,95%CI(0.12,0.33),P<0.00001];shorten the healing time of moderate and severe dermatitis and accelerate the wound healing[MD=-2.56,95%CI(-3.34,-1.79),P<0.00001].The effective rate of mucosal injury treatment of "Kui-Yang-Gel"was significantly higher than that of conventional treatment[OR=3.09,95%CI(1.03,9.27),P<0.05];and the effective time was significantly shorter than that of conventional treatment[MD=-1.86,95%CI(-3.42,-0.30),P<0.05],and the complete pain relief rate was significantly improved[or=4.17,95%CI(1.63,10.67),P<0.01].2 Comparative study of RIOM model:the survival rate of rats in each group was 100%.Compared with the dose of 18Gy,30Gy-dose irradiation can effectively induce severe radiationinduced oral mucositis.The peak of inflammation appears on the 7th day after radiation.At this time,the macroscopic OMI is 4.67±0.50,which is significantly higher than 2.33±0.52 of 18Gy group(P<0.001).Histopathological examination showed extensive mucosal ulcer with a large number of inflammatory cell infiltrating.The body weight of rats decreased by 11.70%±2.00%compared with that before irradiation.3 Comparative study of CTOM model:the survival rate of rats in each group was 100%.Compared with IP group,IP+CS group and Ⅳ group,IP+ MS group can induce severe oral mucositis.The peak of inflammation is on the 4th day.At this time,the macroscopic OMI is 3.50±0.55,which is higher than that of other groups.The microscopic OMI was 3.67±0.58,which was higher than that of other groups.Compared with the control group,immunohistochemical staining showed higher expression of IL-6 and TNF-α,and the difference was statistically significant(P<0.001,P<0.0001).The weight of IP+MS group reached the lowest value on the 5th day,and then gradually recovered.4 The study of "Kui-Yang-Gel" in the treatment of RIOM model rats:Compared with the Control group,the weight of Model group decreased significantly,and the weight of group KFX and NJ was higher than that of Model group.On the 7th day of the experiment,the increasing rate of body weight of Model group was significantly lower than that of control group(P<0.001).Compared with Model group,the increasing rate of body weight of KFX group was higher than that of Model group(P>0.05),and NJ group significantly higher than that of Model group(P<0.01).Compared with the Control group,the Model group suffered from severe oral mucositis.The peak of inflammation in each group occurred on the 7th day of the experiment.At this time,the macroscopic OMI in Model group was 4.67±0.50 and that in KFX group was 4.00±0.71,which was lower than Model group(P>0.05).The NJ group was 3.00±0.71,which was significantly lower than that of the Model group(P<0.001).Histopathological observation showed extensive mucosal loss,massive inflammatory cell infiltration and small abscess formation in Model group.Smaller area of mucosal loss,massive inflammatory cell infiltration was observed in KFX group.And a small area of mucosal loss,less inflammatory cell infiltration,and mild lesion was observed in NJ group.16sRNA sequencing:compared with Control group,the abundance of Gram-positive bacteria decreased and the abundance of Gram-negative bacteria increased in Model group;The species diversity of oral flora decreased significantly.Pathway enrichment suggests a correlation with the colonization,endotoxin synthesis,activation of innate immune system and proinflammatory pathways.And "Kui-Yang-Gel" can inhibit this trend.Transcriptome sequencing:Compared with Control group,the transcription level of TLR4,MyD88 and NF-κB p65 increased in Model group(P>0.05,P<0.05,P<0.05).Compared with Model group,the transcription level of TLR4,MyD88 and NF-κB p65 was lower in NJ group(P<0.05,P<0.05,P>0.05).Western blotting:Compared with Control group,the expression of TLR4,MyD88,TRAF6 and NF-κB p65 in Model group increased significantly(P<0.001,P<0.05,P<0.01,P<0.001).Compared with Model group,the expression TLR4,MyD88,TRAF6,NF-kB p65 of NJ group decreased significantly(P<0.01,P<0.05,P<0.05,P<0.01).Protein quantitative chip:Compared with Control group,IFN-γ(P<0.05)、TNF-α(P<0.05)、IL-1β(P<0.05)、MCP-1(P<0.01)、IL-6(P<0.01)、IL-13、IL-lα、IL-2 and IL-4(P>0.05)was higher in Model group,while the expression of IL-10(P>0.05)was lower.Compared with Model group,the expression of IFN-γ(P<0.01)、IL-1β(P<0.05)、IL-2(P<0.01)、IL-4(P<0.05)、IL-6(P<0.05)、TNF-α、MCP-1、IL-13、IL-1α(P>0.05)was lower in NJ group than that in Model group,and IL-10(P>0.05)was higher than that in Model group.5 The study of "Kui-Yang-Gel" in the treatment of CTOM model rats:Compared with the Control group,the weight of Model group decreased significantly,and the weight of group KFX and NJ was higher than that of Model group.On the 7th day of the experiment,the increasing rate of body weight of Model group was significantly lower than that of Control group(P<0.001).Compared with Model group,the increasing rate of body weight of KFX group was higher than that of Model group(P>0.05),and NJ group significantly higher than that of Model group(P<0.01).Compared with the Control group,the Model group suffered from severe oral mucositis.The peak of inflammation in Model group occurred on the 5th day of the experiment.At this time,the macroscopic OMI in Model group was 4.00±0.50 and that in KFX group was 4.00±0.71 on the 4th day,which was lower than Model group(P>0.05).The NJ group was 3.00±0.71 on the 4th day,which was significantly lower than that of the Model group(P<0.05).The macroscopic OMI in Model group on the 7th day was 3.22±0.67 and that in KFX group was 2.44±0.53,which was lower than Model group(P>0.05).The NJ group was 2.11±0.33,which was significantly lower than that of the Model group(P<0.01).As for microscopic observation,the OMI of Model group was 3.56±0.53,and 2.89±0.78 of KFX group and 2.33±0.50 of NJ group.The microscopic OMI of KFX group was lower than Model group and the difference was not statistically significant(P>0.05).The microscopic OMI of NJ group was lower than Model group and the difference was statistically significant(P<0.01).16sRNA sequencing:Compared with Control group,the abundance of Gram-positive bacteria decreased and the abundance of Gram-negative bacteria increased in Model group;The species diversity of oral flora changed significantly.Pathway enrichment suggests a correlation with the colonization,endotoxin synthesis,activation of inflammatory reaction.And "KuiYang-Gel" can inhibit this trend.Transcriptome sequencing:Compared with Control group,the transcription level of NFκB p65 increased in Model group(P>0.05),and the level of Wnt2 decreased significantly(P<0.001).Compared with Model group,the transcription level of NF-κB p65 decreased in NJ group significantly(P<0.05),and the level of Wnt2 increased significantly(P<0.05).Western blotting:Compared with Control group,the expression of phospho-β-catenin、NF-κB p65 in Model group increased significantly(P<0.05,P<0.05),and the expression of Wnt2 decreased significantly(P<0.05).Compared with Model group,the expression of phospho-β-catenin、NF-κB p65 in Model group decreased significantly(P<0.05,P<0.05),and the expression of Wnt2 increased significantly(P<0.05).Protein quantitative chip:Compared with Control group,IFN-γ(P<0.05)、IL-1β(P<0.05)、IL-6(P<0.01)、MCP-1(P<0.01)、TNF-α(P<0.05)、IL-1α、IL-2、IL-4、IL10、IL-13(P>0.05)was higher in Model group.Compared with Model group,the expression of IFN-γ(P<0.05)、TNF-α(P<0.05)、IL-1α(P<0.05)、IL-6(P<0.05)、IL-1β、MCP-1、IL-13、IL-2(P>0.05)was lower than that in Model group,and IL-4、IL-10 was higher than that in Model group.Conclusion:"Kui-Yang-Gel" can effectively improve radiation-induced dermatitis and mucositis in clinic.A single dose of 30-Gy radiation on the rat snout can effectively construct the severe RIOM model.Intraperitoneal injection of fluorouracil combined with mechanical stimulation of buccal mucosa can effectively construct the severe CTOM model.In the rat model,it can effectively improve RIOM and CTOM.In the RIOM rat model,"Kui-Yang-Gel" inhibits the structural and functional changes of oral flora caused by radiation,and inhibits TLR4-MyD88TRAF6-NF-κB pathway,so as to inhibit the increase of proinflammatory factors.In the CTOM model,"Kui-Yang-Gel" can inhibit the change of oral flora caused by chemotherapeutic drugs and up-regulates Wnt2/β-catenin pathway and inhibits NF-κB from being activated and inhibits the increase of multiple proinflammatory factors. |
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