| ObjectiveTo investigate the distribution of cold and heat syndromes in the active and remission phases of ulcerative colitis(UC),and the possible relationship between the expression characteristics of temperature receptor TRPs at different stages of UC and the cold and heat syndromes in traditional Chinese medicine.Method1.Use computer to search the literature reports on UC TCM clinical research on China Knowledge Network(CNKI),Wanfang Data Knowledge Platform,VIP Database(VIP),PubMed,Web of Science in the past 10 years,after diagnosis criteria,inclusion and exclusion After standard screening,select the literature data that meets the research,and use mathematical statistics to perform statistics and evaluation on the distribution of syndrome types in the clinical research literature of UC during the active and remission periods of TCM treatment,so as to obtain the TCM syndrome types of UC distribution.2.On the basis of the conclusions of the first part of the literature,carry out the second part of the animal experiment research.The classic rat UC model was prepared by the free drinking method of dextran sodium sulfate(DSS)solution,and the control group was free to drink distilled water instead of DSS solution for modeling.In both groups,rat colon tissues were collected 1 day before modeling and 10,17 and 24 days after modeling.Record the general life conditions of rats,such as appearance,activity ability,appetite,stool,etc.Observe and detect the immunohistochemical staining of various temperature receptors(TRPV1,TRPV2,TRPV3,TRPV4,TRPA1,TRPM8)in colon tissue,and routinely perform intestinal tract Pathological inflammatory activity score(Geboes score),combining the general performance of the rat and the general performance of the colon,and clarifying the nodes of the early inflammation,the late inflammation,the remission phase and the healing phase corresponding to the rat UC model.On this basis,immunohistochemical staining was used to determine the spatial distribution and protein expression characteristics of different types of temperature receptors in colon tissue at different disease stages.ResultPart 1:1.According to statistics,a total of 224 documents have been included,involving 15,304 cases.There are 42 types of syndromes,including combined syndromes and composite syndromes.After splitting the combined syndromes and composite syndromes and uniformly naming them,a total of 25 types have been obtained.2.Among all the 25 types of syndromes included,the frequency of occurrence of each type in 224 documents is sorted from high to low.The top 3 types are large intestine damp-heat,damp-heat internal accumulation and spleen and kidney yang deficiency.Among them,the frequency of large intestine damp-heat and damp-heat internal accumulation during the active period was the highest,and the two accounted for 51.42%;the spleen-kidney-yang deficiency syndrome occurred the highest in the remission period,accounting for 50%of all literatures in the remission period.Among all 25 types of syndromes included,the number of cases in each type is sorted from highest to lowest.The top 5 syndrome types are damp-heat in the large intestine,internal accumulation of damp-heat,spleen and kidney yang deficiency,spleen deficiency damp-heat,and spleen deficiency.Dampness accumulation syndrome;the distribution of active stage syndrome types is ranked first in the large intestine damp-heat type,and the disease location syndrome factors are as follows:spleen>large intestine>kidney>liver>stomach>intestinal collaterals according to the number of cases.The number of cases in descending order is:damp>heat>qi deficiency>yang deficiency>stasis>qi stagnation(stagnation)>cold>toxin>yin deficiency>blood deficiency;the distribution of syndrome types in the remission stage is spleen and kidney yang Deficiency is predominant,disease location syndrome factors are as follows according to the number of cases:spleen>kidney>lung>stomach>liver,disease syndrome factors according to the number of cases are as follows:Yang deficiency>qi deficiency>damp>heat>toxic>stasis>qi depression.Part 2:Rats can successfully model the active stage of UC disease by drinking 4.5%DSS solution freely for 10 days;giving 1%DSS solution to continue to drink freely for 7 days,the pathological morphology of rat colon tissue conforms to the late stage of UC inflammation(though there are typical manifestations of UC in the mucosal layer,but inflammatory cell infiltration is less than before);4.5%DSS solution for 10 days+1%DSS solution for 7 days+distilled water for 7 days to drink freely can successfully model the pathological model of UC in remission.The expression level and spatial distribution of 6 temperature receptors(TRPV1,TRPV2,TRPV3,TRPV4,TRPA1 and TRPM8)in colon tissue of UC rats were determined by immunohistochemical analysis,which confirmed that there are 6 temperature receptors in each layer of colon tissue with brush border colonic epithelial cells,goblet cells,lamina propria cells and ganglia expressing more.Changes in the expression of 6 temperature receptors:TRPV1:There is no significant statistical difference between the baseline and the control points of the control group(P>0.05);compared with the corresponding control group,the D10 model group is slightly lower than the control group(P>0.05),the D17 model group was significantly lower than the control group(P<0.05);the D24 model group was slightly higher than the control group(P>0.05);the D10,D17and D24 node model groups had statistical differences(P<0.01),showing a trend of first decline and then rise.TRPV2:There is a statistical difference between the baseline and the control group's detection points(P<0.05);compared with the corresponding control group,the D10 model group is significantly up-regulated than the control group(P<0.05),and the D17 model group was significantly down-regulated than the control group(P<0.01),and the D24 model group was significantly up-regulated compared with the control group(P<0.01);compared with the baseline,the D10 model group was up-regulated(P>0.05),and the D17 model group was significantly down-regulated(P<0.05),the D24 model group was up-regulated(P>0.05);the D10,D17 and D24 nodal model groups were statistically different(P<0.01),showing a trend of first decline and then rise.TRPV3:There is a statistical difference between the baseline and the control points of the control group(P<0.01).Compared with the corresponding control group,the D10 model group was significantly down-regulated compared with the control group(P<0.01),the D17 model group was significantly up-regulated compared with the control group(P<0.01),and the D24 model group was significantly up-regulated compared with the control group(P<0.01);Compared with the baseline,the D10 model group was down-regulated(P<0.05);D17 and D24 model groups were significantly up-regulated(P<0.01);D10,D17 and D24 model groups had statistical differences(P<0.01),the expression value of each node model group shows an upward trend.TRPV4:There is no significant difference between the baseline and the control points of the control group(P>0.05).Compared with the corresponding control group in each node model group,the D10,D17and D24 model groups are all slightly up-regulated than the control group(P>0.05);the expression value of each node model group in D10,D17 and D24 shows an upward trend,but the three are compared There was no statistical difference(P>0.05).TRPA1:There is a statistical difference between the baseline and the control points of the control group(P<0.01).Compared with the corresponding control group,the D10 model group was significantly up-regulated compared with the control group(P<0.01),and the D17 and D24 model groups were slightly up-regulated compared with the control group(P>0.05);each node model group was compared with the baseline,D10 model group was significantly up-regulated(P<0.01),D17 model group was slightly up-regulated(P>0.05);D24 model group was significantly up-regulated(P<0.01);D10,D17,D24 node model groups were statistically different(P<0.01),showing a trend of first decline and then rise.TRPM8:There is a statistical difference between the baseline and the control group's detection points(P<0.05);compared with the corresponding control group,the D10 and D17 model groups are slightly lower than the control group(P>0.05),and the D24 model Compared with the control group,the group was significantly down-regulated(P<0.01);compared with the baseline,the D10 model group was slightly up-regulated(P>0.05);the D17 model group was slightly down-regulated(P>0.05);the D24 model group was significantly down-regulated(P<0.05);each node model group of D10,D17and D24 showed a trend of first decline and then rise,while the D17 and D24 model groups were lower than the D10 model group,and there was no statistical difference between the three(P>0.05).Conclusion1.In the active and remission phases of UC,the main syndrome types are damp-heat of the large intestine and yang deficiency of the spleen and kidney,which have the characteristics of the transformation of cold-heat syndromes at different disease stages.With the transformation of UC syndrome differentiation stage,the expression of temperature receptors changes regularly.In the damp-heat stage of the large intestine(D10-D17),the expressions of TRPV1,TRPV2,TRPA1 and TRPM8 are down-regulated while the expressions of TRPV3 and TRPV4 are up-regulated;In the virtual stage(D17-D24),the expression of temperature receptors showed an upward-regulated trend.The expression characteristics of temperature receptors may have a certain correlation with the conversion of cold-heat syndromes in UC disease.Whether the expression changes of temperature receptors are the result or the cause of cold-heat syndromes remains to be further studied.2.The DSS modeling method has a good dose-time-pathological correlation,and is more suitable for TCM research related to UC staging. |
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