RESUMEN
OBJECTIVE To evaluate the intervention effect of Caulis sinomenii compatible with prepared Aconiti Lateralis on bone destruction in rheumatoid arthritis （RA）model rats ，and to investigate its mechanism. METHODS Totally 40 SD rats were randomly divided into blank group ，model group ，positive control group （indomethacin 0.013 5 g/kg）and C. sinomenii compatible with prepared Aconiti Lateralis group （C. sinomenii 1.08 g/kg+prepared Aconiti Lateralis 1.35 g/kg）according to body mass ，with 10 rats in each group. Except for the blank group ，all the other groups made RA rat models by injecting type Ⅱ bovine collagen. Rats in each group were given corresponding drugs or distilled water intragastrically. The general information ，body weight ，foot swelling and arthritis index （AI）scores of rats in each group were recorded. After the 30th day of administration ，the changes of ankle bone in rats were detected by small animal CT machine. The levels of inflammatory factors [interleukin- 31（IL-31），IL-25 and IL- 3] and chemokines [receptor activator of nuclear factor κB ligand（RANKL），receptor activator of nuclear factor κB （RANK）and osteoprotegerin （OPG）] in serum were detected by enzyme-linked immunosorbent assay. Pathological indexes of rat ankle joint were observed by HE staining. Immunohistochemical method was used to detect the expression of RANKL ，RANK and OPG in synovial tissue of rat ankle joint. RESULTS Compared with blank group ，the mental state of the model group was weak ， the activity decreased significantly ，the hair lost luster ，and the body weight decreased significantly on the 12th to 30th days （P＜ 0.05 or P＜0.01）；the swelling degree of the foot was significantly increased and the AI score was significantly increased on the 12th to 30th days（P＜0.01）；the ankle joint in model group had rough surface ，obvious tissue damage and serious bone erosion ； serum levels of IL- 31，IL-25，IL-3，RANKL and RANK were increased significantly ，while the level of OPG was decreased significantly （P＜0.01）； the expression of RANKL and RANK in synovium of ankle joint increased significantly ， while the expression of OPG decreased significantly （P＜0.01）. Compared with model group ，the above indexes of administration groups were improved to varying degrees ，and most of the differences were statistically significant （P＜0.05 or P＜0.01）. CONCLUSIONS By inhibiting the RANKL/RANK/OPG signaling pathway ，C. sinomenii compatible with prepared Aconiti Lateralis can inhibit the excessive proliferation of osteoclasts and restore the balance of bone metabolism so as to play a role in protecting bone joints and treating RA.
RESUMEN
Objective To explore the effect of sinomenine on the expression of tumor necrosis factor-α (TNF-α),cathepsin G (Cat-G) and cathepsin S (Cat-S) in rats with collagen induced arthritis (CIA).Methods The 60 SPF male Wistar rats were randomly divided into normal group,model group,high dose group,medium dose group,low dose group,and dexamethasone group (with 1 0 in each group).In the normal control group,the rats were given ordinary feed.For the other five groups,the rats were used to build a CIA model and give pharmacologic intervention in the following 20 days.After 20 days of inflammation,sinomenine would be divided into three different dose groups,with 120 mg/kg,90 mg/kg and 60 mg/kg,respectively,and each group was given a gavage daily.For the dexamethasone group,7.5 mg/kg dexamethasone was given for gavage once a day.In terms of the model group and normal group,the rats were perfused with the same volume of saline once daily.Then,taken the pictures of foot paw X-ray and foot paw pictures of rats in each group,measured the content of TNF-α,Cat-G,Cat-S in blood serum,observed the pathological changes in the synovial tissue of rats in each group by tissue section,measured the content of TNF-α,Cat-G,Cat-S in in spleen of rats by Immunohistochemical staining.Results The results of X-ray showed that there were obvious soft tissue swelling,joint deformity and osteolysis in the paws of the model group,and the above symptoms were alleviated in different degrees in each treatment group compared with the model group.Compared with the model group,the expression of TNF-α (376.48 ± 22.21 pg/ml,369.45 ± 82.68 pg/ml,425.17 ± 153.51 pg/ml vs.457.63 ± 152.67 pg/ml),Cat-G (1 398.05 ± 167.32 pg/ml,1 337.65 ± 209.34 pg/ml,1 412.78 ± 67.65 pg/ml vs.2 283.03 ± 185.21 pg/ml),and Cat-S (662.18 ± 169.66 pg/ml,406.80 ± 41.93 pg/ml,452.76 ± 50.49 pg/ml vs.838.11 ± 141.86 pg/ml) in blood serum of sinomenine high dose group,medium dose group and low dose group significantly decreased (P＜0.05).The expression of TNF-α (0.28 ± 0.05,0.21 ± 0.03,0.34 ± 0.04 vs.0.50 ± 0.04),Cat-G (0.28 ± 0.02,0.18 ± 0.06,0.27 ± 0.02 vs.0.37 ± 0.03),and Cat-S (0.22 ± 0.02,0.18 ± 0.03,0.27 ± 0.02 vs.0.35 ± 0.03) in spleen tissue significantly decreased (P＜0.05).The results of HE staining showed that the synovial tissue of normal rats was normal,the synovial tissue cells of model rats were damaged,the expression of inflammatory cells was significantly increased,and pannus hyperplasia was observed.Inflammatory cell infiltration and pannus hyperplasia were decreased in each group after administration.Conclusions Sinomenine has a sound and comprehensive intervention effect on rheumatoid arthritis,and the mechanism may be related to the inhibition of the expression ofTNF-α,Cat-G and Cat-S.
RESUMEN
This study was aimed to improve the transfer rate of sinomenine in the intermediate product of Caulis Sinomenii in order to optimize the purification process of intermediate product ofCaulis Sinomenii fromTong-An (TA) injection. The transfer rate of sinomenine and the stability of fingerprints in the intermediate product of Caulis Sinomenii were used as indexes for the investigation on the impact from different pH ofCaulis Sinomenii extract before extraction. The transfer rate of sinomenine was used as an index for the investigation on the impact by different pH of hydrochloric acid created to dry extract solution. The transfer rate of sinomenine was used as an index for the investigation of four separation ways, which included the vacuum filtration, plate and frame filters, high-speed tube separator, and flat direct centrifuge, on the liquid separation of sinomenium acutum acid. The results showed that the pH ofCaulis Sinomenii extract before extraction was 10-11; the transfer rate of sinomenine was the highest in the extraction process and the fingerprints of TA injection was stable. The pH of hydrochloric acid was 2.0-2.5; and the highest transfer rate of sinomenine in acid dissolution process was 92.94%. The high-speed tube separator had the best separation to sinomenium acutum acid-dissolving liquid. The highest transfer rate of sinomenine was 93.34%. It was concluded that the optimized process can effectively improve the transfer rate of sinomenine in the intermediate product ofCaulis Sinomenii. Meanwhile, fingerprints of the product were stable. The process was simple with good repeatability.