Effects of swertiamarin bitter glycoside and Iingxian new glycoside component pairing on serological indices and ankle COX-2 in RA model rats
Objective Investigating the effects of swertiamarin and lingsenoside fractions on serological indices and ankle COX-2 in a rheumatoid arthritis(RA)model rat.Methods Thirty-six SPF-grade SD rats of 5-6 weeks of age,half male and half female,were selected.Except for the blank group,the rat model of Collagen induced arthritis(CIA)was replicated using the type Ⅱ collagen induction method.The 30 rats successfully modeled were divided into the model group,the positive drug(Zhengqing Fengqin)group,the swertia bittersweet group,the lingxian xinsenoside group,and the swertia bittersweet with lingx-ian xinsenoside group(hereinafter referred to as the"swertia-lingxian group")according to the method of randomized numerical tables,and there were 6 rats in each group.The drug groups were given the corresponding liquid by gavage,and the model group and the blank group were given an equal amount of saline by gavage for 10 d.In the experiments,the general state of the rats in each group was observed and recorded,arthritis scores(AI)were recorded,and histopathological changes of the ankle joints of the rats were observed by hematoxylin-hematoxylin(HE)staining method;the serum tumor necrosis factor-α(TNF-α)and the tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA);the serum tumor nec-rosis factor-α(TNF-α)and tumor necrosis factor-α(TNF-α)were measured by ELISA.Tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1β),rheumatoid factor(RF),C-reactive protein(CRP),anti-cyclic citrullinated peptide an-tibody(anti-CCP Ab)and prostaglandin E2(PGE2)in rat serum were detected by enzyme-linked immunosorbent assay(ELISA),and rat ankle joint cyclo-oxidation was detected by immunohistochemistry and protein immunoblot(Western blot).The protein expression of rat ankle cyclooxygenase-2(COX-2)was detected by immunohistochemistry and Western blotting;the gene expression of rat ankle COX-2 was detected by real-time PCR.Results Compared with the blank group,the general state of rats in the model group was worse,and the AI score and serum levels of TNF-α,IL-1 β,RF,CRP,anti-CCP Ab,and PGE2 were significantly higher than those in other groups(P＜0.05 or P＜0.01)in the left hind toe of rats;the ankle joint tissues and structures were severely damaged,and the expression of COX-2 proteins and genes were significantly up-regulated(P＜0.01).Compared with the model group,the general state of rats in each dosing group showed different degrees of improve-ment;Al scores were significantly reduced(P＜0.01),and serum levels of TNF-α,IL-1 β,RF,CRP,anti-CCP,and PGE2 were significantly reduced(P＜0.05 or P＜0.01),among which the swertia-ling dosing group inhibited TNF-α,IL-1β,RF,CRP,anti-CCP and PGE2 were the most prominent;the histopathological status of ankle joints was improved to differ-ent degrees;IHC and WB test results showed that the swertia-spirit combination group reduced the protein expression level of COX-2 in ankle joints most significantly(P＜0.01);and PCR results showed that swertia-spirit ascorbic acid group reduced the gene expression of COX-2 in ankle joints most significantly(P＜0.01).Conclusion Swertia bittersweet,lingxian xinoside and their combination can play a good therapeutic effect on RA model rats,in which the therapeutic effect of the swertia-lingxian combination group was significantly better than that of the monomer administration group;swertia bittersweet and lingxian xinoside have obvious synergistic effect,and their mechanism of action may be to play a role in the reduction of the secretion of inflammato-ry cytokines,and the inhibition of the activity of the COX-2 protein.
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