The Research On The Relationship Of Intestinal Flora Disturbance And The Pathogenesis Of Eczema In Mice

Objective: By detecting the skin barrier function, serum levels of IFN-γand IL-4, and blood CD4, CD8levels in mice to explore the intestinal floradisturbance caused mice skin eczema mechanisms and the role of probiotics.Methods:1. Grouping method: choose90SPF kunming mice, male andfemale half, feeding the adaptability in mice after5d randomly，divided into3groups, namely the normal control group (group A), model group (group B),gold dichotomous treatment group (group C),30in each group.2. Tests in miceback skin water loss (TEWL) and percutaneous corneous layer water content bymulti-functional skin tester MPA9made in German Courage Khazakaelectronic Company; Test environment temperature is19.8℃~21.4℃, relativehumidity is45.8%~49%. The basic unit of corneous layer water content and thelipids of skin is au (arbitraryunit), percutaneous moisture loss amount of the unitis g/m2h.3. Use the method of direct smear feces specimen observationdetection of intestinal flora: Fresh manure0.1g after adding0.9ml of sterilesaline and homogenized to break the bead solution, suspension formed stool smears taken by Coriolis improved method for staining Gram staining stainingsuccessfully observed directly using oil immersion stool specimens situation.4.Using SemiBio cell counting slide were detected in mouse blood CD4, CD8levels and CD4/CD8.5. Detection of mice serum IFN-γ, IL-4cytokine levels bythe ELISA method.6. The resulting data using SPSS13.0statistical software forstatistical analysis, and the resulting measurement data for mean±standarddeviation (±s) said, using the t test, analysis of variance for statistical analysisto P<0.05was considered statistically significant.Results: The model group compared with the control group, significantlyreduced the total number of intestinal flora, but each of the three groups of miceintestinal flora bacteria count of the total number of rate change, the resultsshow: Group B compared with group A and group C, G+b and G-b number oftotal bacteria in the intestine was significantly down, G+c and G-c accounts forthe ratio of the total number of bacteria increased significantly, the differenceswere statistically significant (P <0.01); with group A compared to group C, G+bratio of the total number of bacteria increased, the difference was statisticallysignificant (P <0.05), while the rest of the flora of the total number of bacteriawas no significant difference in the number of ratios.TEWL and water content of the stratum corneum of skin test results in mice:Compared with group A TEWL value in mice, B and C groups was statisticallysignificant (P <0.05); compared with group B TEWL values, no statisticaldifference in group C significance (P>0.05); group B corneous layer watercontent compared with group A and C, significant difference was statisticallysignificant (P <0.01); group A compared with group C, the difference was not statistically significant (P>0.05).Mouse blood CD4, CD8expression level results show: Group B comparedwith group A and group C, degree of CD4, CD8expression is suppressed, have astatistically significant difference (P<0.01); compared with group A, C groupsCD4, CD8expression was reduced, the difference was statistically significant.(P <0.05).Mouse serum IFN-γ, IL-4cytokine levels showed that: Compared withgroup A,IL-4elevated levels of group B, lower IFN-γ levels, the differenceswere statistically significant (P<0.05); compared with group B, group C IFN-γlevels increased, the difference was statistically significant (P<0.05), whereasIL-4levels showed no significant difference (P>0.05); group A compared withgroup C, the difference was not statistically significant (P>0.05).HE staining light microscopy showed that after2weeks with antibioticsorally, part B and C group mice back and rashes on the anus week samplechange, light microscope epidermal parakeratosis, epidermal local necrosis,exudation and accompanied inflammatory cell infiltration, infiltration oflymphocytes and a few cells were mainly neutrophils, in line with thepathological changes of acute eczema. Group C mice fed with Jinshuangqi10dafter the rash has improved, optical microscope, subacute eczema andconvalescent changes to the transformation of chronic eczema, showedhyperkeratosis, parakeratosis, granular layer thickening, mild skin hypertrophy,superficial dermal edema with fibroblasts, basal cell liquefaction has beenobvious. Conclusion:(1) The intestinal flora disturbance induced by antibiotics can make skinrashes in mice skin rash.(2) Intestinal flora disorder can cause IFN-γ, IL-4in mice serum cytokineimbalance and the decrease in the number of CD4, CD8cells, which may berelated to mouse skin eczema have a certain relationship.(3) Intestinal disorders cause skin eczema increased with the percutaneousmoisture loss and corneous layer water content reduce also to have certainrelations.(4) Probiotics to the antibiotic cause the regulation of intestinal floradisorder have obvious effect, and skin eczema has certain therapeutic effect inmice.The conclusion of this experiment can be used probiotic preparations forthe prevention and treatment of allergic skin diseases in the future to provide atheoretical basis for clinical treatment.