中國實(shí)驗(yàn)方劑學(xué)雜志 第29卷 第11期 2023年6 月
［Abstract］ Objective： To investigate the anti-inflammatory effect of the component compatibility of
Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma on the rat model of rheumatoid arthritis （RA） and the mechanism. Method： Seventy-two SPF-grade SD rats （male and female） aged 5 to 6 weeks were selected. Except the blank group， the rat model of collagen-induced arthritis （CIA） was replicated by the type Ⅱ
collagen induction method. The 64 rats after successfully modeling were randomly divided into model group，
methotrexate group （0.375 mg·kg-1）， gentianoside with magnoflorine group （150.454 1 mg·kg-1+5.061 8 mg·kg-1）， gentianoside with clematichinenoside AR group （150.454 1 mg·kg-1+16.433 1 mg·kg-1）， sweroside with magnoflorine group （3.455 8 mg·kg-1+5.061 8 mg·kg-1）， sweroside with clematichinenoside AR group （3.455 8 mg·kg-1+16.433 1 mg·kg-1）， swertiamarin with magnoflorine group （9.303 2 mg·kg-1+5.061 8 mg·kg-1）， and swertiamarin with clematichinenoside AR group （9.303 2 mg·kg-1+16.433 1 mg·kg-1）， with 8 rats in each
group. Each group was given the corresponding medicinal solution or normal saline by gavage for 15 d. During
the experiment， the general status， of rats in each group were observed and recorded. Tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， rheumatoid factor （RF）， C reactive protein （CRP）， prostaglandin E2 （PGE2）， and anti cyclic peptide containing citrulline antibody （anti-CCP Ab） in the serum of rats were measured by enzyme linked immunosorbent assay （ELISA）. The histopathological changes in rat ankle joints were observed by hematoxylin-eosin （HE） staining. Immunohistochemistry （IHC） and Western blot were used to detect the protein expression of nuclear factor-κB （NF-κB） and vascular endothelial growth factor （VEGF） in rat ankle joints. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA
expression of NF-κB and VEGF in rat ankle joints. Result： Compared with those in the blank group， rats in the model group were in poor general conditions with significant foot-plantar swelling， and the content of CRP， anti-
CCP Ab， and IL-1β in the rat serum was significantly increased （P<0.01）. In the model group， the tissue
structure of the ankle joint was severely damaged， and the protein and mRNA expression of NF-κB and VEGF in
the rat ankle joints were significantly up-regulated （P<0.01）. As compared with the model group， the general
status of rats in each administration group was significantly improved. The levels of serum TNF-α， IL-1β， RF， CRP， PGE2， and anti-CCP Ab were reduced to different degrees in these administration groups， among which the effects of the gentianoside with clematichinenoside AR group on down-regulating serum TNF-α and IL-1β， the gentianoside with magnoflorine group on down-regulating serum RF and CRP， the sweroside with magnoflorine group on down-regulating serum PGE2， and the swertiamarin with clematichinenoside AR group on lowering serum anti-CCP Ab were better than those of administration groups. The histopathological changes in
the ankle joint were improved to different degrees. The protein and mRNA expression of NF-κB and VEGF in rat
ankle joints in the administration groups was significantly down-regulated （P<0.05， P<0.01）， and the
swertiamarin paired with clematichinenoside AR group had the most significant effect. Conclusion： The component compatibility of Gentianae Macrophyllae Radix and Clematidis Radix et Rhizoma exerts a good therapeutic effect on the rat model of RA， and the compatibility of components from the two medicines has a multi-channel， multi-target， and synergistic effect. The five component compatibility patterns， namely gentiobioside with magnoflorine， gentiobioside with clematichinenoside AR， sweroside with clematichinenoside AR， swertiamarin with magnoflorine， and swertiamarin with clematichinenoside AR， all have potential advantages. The mechanism may be related to the reduction of inflammatory factor secretion and the inhibition of
abnormal protein and mRNA expression of NF-κB and VEGF.
本研究中對照品：靈仙新苷AR 購自于四川恒誠致遠(yuǎn)生物科技有限公司（Naturewill biotechnology Co., Ltd.）
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