| Objective: We developed the CIA model rats and gave different doses of ethanol extract from pterocarya hupehensis skan.Finally we would through the detection of IL-1?,TNF-? and NF-?B phosphorylation p65(P-p65),Cleaved Caspase-3 and other indicators to explore the effect of the drug on the CIA rats and possible mechanisms.Methods:(1)60 male SD rats(weight 200 ± 20g)were randomly divided into negative control group,model group,Pterocarya hupehensis Skan low,medium,high dose group and positive control group.Each group has 10 rats.In addition to the negative control group,the rest shall be built the CIA model.After it was established,each group would give corresponding drug dose,negative and model groups were given equal volume saline,rats executed after 28 days of continuous administration.(2)We will observe and record the general conditions of rats,including rats 'weight,arthritis index,joint swelling.We 'll have a pathological examination of the synovial tissue of the left knee.Tunel method was used to detect apoptosis of synovial cells in rats.ELISA method was used to detect the levels of il-1? and TNF-? in rat serum.Western Blot method was used to detect the protein content of P-p65 and Cleaved Caspase-3 in right knee synovial tissues of rats.The levels of ALT,AST,SCr and BUN were detected in serum of rats.Results:(1)General observations: The results showed that compared with the negative control group,the weight of rats in the model group decreased,but the joint swelling degree and arthritis index reduced.The results of the high dose group were slightly better than the positive group(p < 0.05).(2)Observation on the pathological morphology of the synovial tissue: The synovial cells in the synovial tissue of the model group were increased,and the synovial lining layer was proliferated,thickened,disordered and evacuated.A large number of inflammatory cell infiltration,visible angiogenesis,even vascular pannus formation.After treatment,the synovial tissue was found to improve in synovial cell proliferation,inflammatory cell infiltration,andvascular pannus,and the high dose group was more effective than the positive control group.(3)Tunel detection: From the optical microscope,we would find compared with the model group,the number of apoptosis was increased in the drug groups,and the high-dose group was significantly higher than that in the positive control group(P<0.05).(4)ELISA detection: compared with the model group,the contents of IL-1?and TNF-? in the serum of each treatment group were decreased obviously,high dose group was better than positive group(P<0.05).The content of ALT,AST,Cr and BUN in the serum was no difference compared with the normal control group.(5)Western Blot method: Compared with the negative control group,the expression of P-p65 in synovial membrane increased.Compared with the model group,the content of P-p65 protein decreased and that of Cleaved Caspase-3 protein increased obviously.The high dose group was slightly better than the positive group(P < 0.05).(6)Biochemical index: The content of ALT,AST,SCr,BUN in serum of model group increased,but the content of drug in pterocarya hupehensis skan group decreased significantly(P < 0.05).But the positive control group did not improve significantly.Conclusion:(1)The ethanol extract from pterocarya hupehensis skan has a good therapeutic effect on CIA model of SD rats.(2)The pterocarya hupehensis skan can play a good role in anti-inflammatory and induction of apoptosis of synovial membrane by inhibiting il-1?,TNF-?,P-p65 and the expression of apoptosis protein Cleaved Caspase 3.(3)The pterocarya hupehensis skan has some protective effect on liver and kidney function. |
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