| Objective:The RA heat syndrome group was higher in the body surface temperature and inflammatory cytokines than the cold syndrome group,but the blood viscosity and fibrinogen in the blood coagulation factor were lower than the cold syndrome group.e syndrome of cold and heat.Methods:Experiment 1st:19 female Wistar rats were randomly divided into 4 groups:the normal group did not intervene,the model group replicated the collagen-induced arthritis(CIA)rat model;the cold syndrome group and the heat syndrome group applied the animal climate chamber based on the replicating CIA model.and were given the wind-cold-dampness and wind-heat-dampness stimulation starting from the initial immune for 28 d.From the baseline period,rat body weight,foot swelling drainage volume,and arthritis index(AI)were measured once a week.From the second week of modeling,the rat joint pain threshold and the peripheral body surface temperature were measured once a week.After 4 weeks of modeling,peripheral blood,ankle joint and heart,liver,spleen,lung,kidney,and thymus were taken.(1)Blood viscosity and coagulation factors were measured by blood viscometer and automatic blood coagulation instrument.(2)Observing the histopathological features of the ankle joint.(3)Calculate each organ index after weighing.Result:The morbidity rate of the three groups was 100%(5/5),and the CIA group was able to induce CIA in 100%(5/5)rats,the CIA cold syndrome group was able to induce CIA in 77.78%(4/5)rats.the CIA heat syndrome group was able to induce CIA in 77.78%(4/5)rats.(1)The body weight of the normal group showed a gradual upward trend.The body weight of the model group,the cold syndrome group and the heat syndrome group decreased after 21 days of modeling,and the difference between the model group and the cold syndrome group and the normal group was statistically significant(P<0.01).There were significant differences between the model group,the cold syndrome group,the heat syndrome group and the normal group after 28 days(P<0.01).(2)The left foot drainage volume of the CIA model group,the heat syndrome group and the cold syndrome group gradually increased during the modeling period,and the left foot drainage volume was higher than the normal group(P<0.01)At 7,14,21,28 days.the right foot drainage volume was higher than the normal group(P<0.01)At 14,21,28 days.The drainage volume of the left foot in the heat syndrome group was higher than that in the model group and the cold syndrome group(P<0.05),and the other time points were not statistically significant(P>0.05).The AI scores of each group showed a gradual increase during the modeling period,but the difference was not statistically significant.(3)On the 14th day,Double foot skin temperatures of three groups were increased to different degrees compared with the normal group(P<0.01),On the 21st day,the bipedal temperature of the model and heat syndrome group was still higher than that of the normal group(P<0.01).The cold syndrome group was lower than the model group and heat syndrome group(P<0.01).On the 28st day,the bipedal temperature in the heat syndrome group was statistically significant compared with the normal group and the model group(P<0.01).the cold syndrome group was lower than other three groups(P<0.01),model group was higher than the normal group(P<0.01).(4)On the 14th day,the pain threshold of the two groups in the model group and the heat syndrome group was lower than that in the normal group(P<0.01 P<0.05).The pain threshold of the left foot in the cold syndrome group was lower than that in the normal group(P<0.01).The pain thresholds of the three groups of double foot were increased in different degrees compared with the normal group at 21 days and 28 days(P<0.01).(5)There was no significant difference in blood viscosity between the four groups at high shear rate(200/s)(P>0.05),the blood viscosity of CIA cold syndrome group was significantly higher than that of the other three groups(P<0.01).At the low shear rate(1/s),the blood viscosity of the heat syndrome group was lower than that of the model group(P<0.05);Fibrinogen concentration(Fib)in CIA cold syndrome group was significantly higher than that in normal group,model group and heat syndrome group(P<0.01).(6)Histopathology showed that no abnormalities in the normal groupthe.In other three groups,the joint cavity was narrow,the focal area disappeared,the synovial cells were moderately hyperplasia,the articular surface was discontinuous,the articular cartilage was destroyed,and the cartilage and the surrounding fibrous connective tissue were adhered,Inflammatory cell infiltration;(7)The thymus index of the heat syndrome group and the cold syndrome group was lower than that of the normal group and model group(P<0.05),and the spleen index of the normal group was lower than that of other groups(P<0.01),cold and heat groups was higher than that in model group(P<0.01).Experiment 2nd:30 female Wistar rats were divided into normal group,cold syndrome group and heat syndrome group.After 4 weeks of modeling,peripheral blood,synovial tissue,were taken.(1)The expression of Egr2,SOCS3,STAT3 and IL-17A in serum of peripheral blood was determined by ELISA.(2)Immunofluorescence assay for the morphological expression of Egr2,SOCS3,STAT3 and IL-17A in the synovium.(3)RT-qPCR was used to determine the expression of LncRNA ZFAS1 mRNA,Egr2 mRNA,SOCS3 mRNA and STAT3 mRNA in the synovium.Result:(1)Peripheral blood results showed that the cold syndrome group,the heat syndrome group Egr2,Socs3,IL17A were significantly higher than the normal group(P<0.01),the heat syndrome group Egr2,IL17A was higher than the cold syndrome group(P<0.01).STAT3 was lower in the cold and heat syndrome groups than in the normal group(P<0.01).(2)LncRNAZFAS1 mRNA and Egr2 mRNA in the heat syndrome group and the cold syndrome group were higher than the normal group(P<0.05 P<0.01),in which the Egr2 mRNA of the heat syndrome group was higher than that of the cold syndrome group(P<0.01),the SOCS3 mRNA of the heat syndrome group was higher than that of the normal group(P<0.05).the STAT3 mRNA of the cold syndrome group and the heat syndrome group was lower than the normal group,There was a statistically significant difference between the heat syndrome group and the normal group(P<0.05).(3)Egr2 was mainly expressed in the nucleus,SOCS3 was mainly expressed in the cytoplasm,STAT3 was mainly expressed in the cytoplasm and nucleus,and IL17A was mainly expressed in the cytoplasm.There was no significant difference in the average fluorescence intensity of Egr2,SOCS3,STAT3 and IL17A in synovial membrane among the three groups(P>0.05).Conclusion1.RA cold syndrome and heat syndrome have their own biological characteristics:The RA heat syndrome group was higher in the body surface temperature and inflammatory cytokines than the cold syndrome group,but the blood viscosity and fibrinogen were lower than the cold syndrome group.2.LncRNA ZFAS1 is involved in the RA disease process but is not associated with the Egr2-mediated inflammatory pathway. |
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