| Objective: This experiment by observing the phlegm dampnessunder the action of CIA mice arthritis index,ankle local tissue of leptin,NLRP3,cleaved-caspase-1,IL-1β,IL-18 pathological and histological differences in changes of protein expression and ankle explore phlegm dampness factors influence on degree of CIA mice ankle joint disease,detection of local tissue leptin levels and NLRP3 inflammatory corpuscle level,To explore whether leptin affects the joint severity of CIA mice through the NLRP3/caspase-1 signaling pathway,and to provide a new theoretical reference for the clinical treatment and pathogenesis of RA.Methods: 32 4-week-old male SPF grade DBA / 1 mice were randomly divided into two groups according to body weight,16 mice in each group.After one week of adaptive feeding,one group continued to be fed with normal diet and the other group was fed with high-fat diet.After 5 weeks of continuous feeding,8 mice were randomly selected from the two groups,and the other group was fed with normal diet and highfat diet.Finally,the mice were divided into normal group,CIA model group,phlegm dampness group and phlegm dampness CIA group,with 8 mice in each group.During the experiment,the general situation of mice was observed and recorded,the Lee’s index of mice was calculated,and the joint inflammation was scored.The pathological changes of ankle joint were evaluated by HE staining,safranin o-solid green staining and micro CT imaging.The serum lipid level was detected by enzyme-linked immunosorbent assay.The protein expression levels of leptin,NLRP3 and cleavedcaspase-1 in ankle joint were observed by Western blot.Results: 1.Compared with the normal group,the mice in the phlegm dampness group were obese,less active,like to pile up,squint,prone to lying,greasy hair,dark and lusterless,stool was not formed,and the phlegm dampness model was established at the age of 10 weeks;the mice in the phlegm dampness CIA group showed more phlegm dampness after CIA modeling.2.There was no significant difference in body weight and Lee’s index between CIA group and normal group(P>0.05).Compared with normal group,the body weight and Lee’s index of phlegm dampness group and phlegm dampness CIA group were significantly increased(P<0.01).3.The levels of serum TC and TG in the phlegm dampness group and the phlegm dampness CIA group were significantly higher than those in the normal group(P<0.01),and were also significantly higher than those in the CIA group,and there was no significant difference in the levels of serum TC and TG between the CIA group and the normal group.4.Compared with CIA group,the AI score of phlegm dampness CIA group was significantly increased(P<0.01).5.Micro CT scanning results showed that there was no obvious bone erosion and destruction in the ankle joint of normal group and phlegm dampness group Results:compared with the phlegm dampness group,the bone erosion and bone destruction of the ankle joint in the phlegm dampness CIA group were more significant;the HE staining results of the ankle joint showed that the local histopathological inflammation score of the normal group was 0,compared with the phlegm dampness group,the local histopathological inflammation score of the ankle joint in the CIA group and the phlegm dampness CIA group was significantly higher(P<0.01),and the phlegm dampness CIA group was significantly higher than the CIA group(P<0.01)The results of safranin osolid green staining showed that the score of ankle cartilage erosion in normal group and phlegm dampness group was 0,and that in CIA group and phlegm dampness CIA group was significantly higher than that in CIA group(P<0.01).6.The expression of NLRP3,pro-caspase-1 and cleaved protein in the local tissues of ankle joint in normal group and phlegm dampness group-Compared with the normal group,the relative expression of NLRP3,pro-caspase-1,cleaved-caspase-1,IL-18,pro-il-1 β and IL-1 βin ankle joint tissues of CIA group and phlegm dampness CIA group was significantly higher(P<0.05),espe CIAlly in phlegm dampness CIA group(P<0.01)The relative expression of leptin,OB Rb and IL-6 in ankle joint tissues of CIA group and phlegm dampness CIA group was significantly higher than that of normal group.Compared with phlegm dampness group,the relative expression of leptin,OB Rb and IL-6 in ankle joint tissues of CIA group and phlegm dampness CIA group was significantly higher,and the phlegm dampness CIA group was significantly higher than that of CIA group(P<0.05).Conclusion: 1.Phlegm dampness can aggravate the degree of arthritis in CIA mice.2. Phlegm dampness can promote the secretion of leptin in the ankle joint of CIA mice.3. eptin may aggravate the inflammatory response of CIA mice by regulating the NLRP3/caspase-1 signaling pathway in the local tissues of ankle joint. |
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