| [Objective ](1) to establish mice model with obesity and periodontitis.(2) to detect the expression of serum inflammatory cytokines such as TNF-?, IL-1?, IL-6 in the model mice.(3) to analyze the effects of periodontitis on obese mice kidney tissue structure and the expression of renal tissue inflammation factor. [Methods]Chapter one(1)There were 44 C57 BL / 6J mice were randomly divided into two groups, one group was feed with high fat food(HF), another group with low fat food(LF).(2) When raised to 16 weeks, the two groups were divided into ligation and sham-ligation groups.the time of ligation was 10 days, and then evolved into four groups: high fat with sham ligation group(HFC), high fat with ligation group(HFP), low fat with ligation group(LFP), low fat with sham ligation group(LFC) named control group.(3)The mice's weight were measured every week. To compare the difference of the body weight of mice in high fat diet group(HF) and low fat diet group(LF) with one-way analysis of variance.(4)After the 10 days of ligation, dealed with all the mice with euthanization and took the maxilla to compare the difference of the ligation and sham ligation.Chapter two Took the sample of all the mice blood, to detect the expression of serum inflammation factors such as TNF-?, IL-1?, IL-6 within the groups of mice with protein chip method.Chapter three(1) Got the mice left and right sides of the kidney tissues and weighed.(2) Detected the m RNA expression of inflammation factor TNF-?, IL-1?, IL-6 in the kidney tissues.(3) Compared the histopathological change of kidney of all the groups with HE and Masson staining. The inflammatory factor IL-1? immunohistochemical staining method, to observe the kidney structure of obesity and periodontitis model mice and the influence of IL-1? expression level of the renal tissue. [Results]Chapter one(1) The weight of mice in high-fat group(HF) and low fat group(LF) had no statistical differences initially(F=0.864, P > 0.864); At the 16 th week, the weight of high-fat group(HF) statistically increased about 13.4 g(43.5%) compared with low fat group(LF)(P< 0.01).(2) After 10 days of ligation, the weight of low-fat group(LF) dropped by about 9.7%(3 g), high fat group(HF) was 16.1%(7.1 g).But the weight of high fat group were still statistically higher than the low fat group(33.5%, 9.3g).(3)The mice of ligation showed gingival papilla atrophy, gingival epithelial root to shift, alveolar crest of absorption, distribution of osteoclasts, but the sham ligation did not.Chapter two(1) Obesity and periodontitis have no interaction on the expression of TNF-?(F=0.061, P=0.061). It was not obesity but periodontitis have the influence of statistical significance on the expression of TNF-? in serum(P<0.001). The level of serum concentrations of TNF-? of ligation group was significantly lower than sham ligation(F=13.6, P<0.001). One-way ANOVA showed high fat with sham ligation group(HFC) was significantly higher than high fat with ligation group(HFP)(P<0.05).(2) For the IL-1?, obesity and periodontitis has no statistical significance interaction(F=1.259, P=1.259). Both obesity and periodontitis had statistical significance effect on the expression of IL-1?(P < 0.05). The IL-1? of ligation group were significantly lower than sham ligation group(C)(F = 5.056, P = 5.056), so did the low fat group(LF) significantly than that the high fat group(HF)(F=15.375, P<0.001). One-way ANOVA showed that high fat with sham ligation group(HFC) were significantly higher than that of low fat with sham ligation group(LFC)(F=11.045, P=0.002), and so it was to high fat with ligation group(HFP)(F=19.948, P<0.001).(3) For the expression of IL-6 in serum, two-way ANOVA showed showed obesity and periodontitis had statistical significance interaction effect(F=19.316,P<0.001)?Obesity and periodontitis both had significant effect on the expression of IL-6 in serum(P<0.05).The ligation group was significantly lower than that of sham ligation group(C)(F=12.4, P=0.001).High fat group(HF)was significantly higher than that of the low fat group(LF)(F=26.837, P < 0.001). One-Way ANOVA showed that the expression of IL-6 high fat with sham ligation group(HFC) was significantly higher than high fat with ligation group(HFP) and low fat with sham ligation group(LFC)(F=16.715,P<0.001.F=22.841,P<0.001.)Chapter three(1) After the mice induced with high fat for16 weeks and ligature for 10 days, Factorial analysis showed obesity and periodontitis had no statistical significant interaction. The mice kidney weight of high fat group(HF)were significantly heavier than of low fat group(F=18.2,P<0.001). T tset showed that the mice kidney weight in high fat without ligation group(HFC) was significantly heavier than of control group(LFC)(F=20.188,P=0.001).(2) HE staining: mice kidney tissues in high-fat group were characterized by void lesion which located in the renal tubular wall. Renal tubular epithelial cell swelling, even serious was the visible renal tubular shape disappeared. mice kidney tissue structure in low fat group(LFP,LFC) were clear, had no obvious structural change.(3)Masson staining: Four groups of mice kidney tissues were not seen obvious collagen deposition.(4)PCR: for the inflammatory markers TNF-?,IL-?, IL-6, obesity and periodontitis had no statistical significant interaction(F=0.659,P=0.427;F=0.494,P=0.491;F=0.434,P=0.519). Factorial analysis showed obesity was not statistically significant on the expression of TNF-?, IL-1?, IL-6. So did the periodontitis except IL-1?.The ligation group was statistically significant higher than the sham ligation. T tset showed that there was no statistical significance between the four groups about the expression of TNF-?, IL-1?, IL-6.(5)IL-1? immunohistochemical staining: low fat group(LFC) was the least on the expression quantity of IL-1? compared with the others. Among them, low fat with ligation group(LFP) renal tissue in the amount of IL-1? expression was thsignificantly higher than the control group(LFC), high fat without ligation group(HFC), high fat with ligation group(HFP) in the inflammatory cytokines IL-1? expression quantity is similar, had no obvious difference. [Conclusion]Chapter one The mouse model of obesity and periodontitis were established successfully.Chapter two Periodontitis may have reduced serum TNF-?, IL-1, IL-6. Obesity may have the effect of stimulation increase serum IL-1?,IL-6.Chapter three(1) Diet-induced obesity might cause hypertrophy in kidney tissue, cavity lesions.(2) Periodontitis may stimulate increase renal tissue inflammation factor IL-1?. |
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