Jinwujiangu Capsule Treats Fibroblast-Like Synoviocytes of Rheumatoid Arthritis by Inhibiting Pyroptosis via the NLRP3/CAPSES/GSDMD Pathway.

Jinwujiangu capsule (JWJGC) is a traditional Chinese medicine formula used to treat rheumatoid arthritis (RA). However, whether its mechanism is associated with pyroptosis remains unclear. In this study, the ability of JWJGC to inhibit the growth of fibroblast-like synoviocytes of RA (RA-FLS) through pyroptosis was evaluated. The cells isolated from patients with RA were identified by hematoxylin and eosin (H&E) staining, immunohistochemistry, and flow cytometry. After RA-FLS were treated with different concentrations of JWJGC-containing serum, the cell proliferation inhibition rate, expression of caspase-1/3/4/5, NOD-like receptor protein 3 (NLRP3), gasdermin-D (GSDMD), and apoptosis-associated speck-like protein containing a CARD (ASC), concentrations of interleukin-1ß (IL-1ß) and interleukin-18 (IL-18), the activity of lactic dehydrogenase (LDH), and pyroptosis were evaluated. The results showed that JWJGC increased the proliferative inhibition rate, decreased the expression of caspase-1/3/4/5, GSDMD, NLRP3, and ASC, suppressed the expression of IL-1ß and IL-18, induced the activity of LDH, and downregulated the number of double-positive FITC anti-caspase-1 and PI. Generally, our findings suggest that JWJGC can regulate NLRP3/CAPSES/GSDMD in treating RA-FLS through pyroptosis.

[Effects of Jinwu Jiangu recipe on IL-17/STAT3 signals in rheumatoid arthritis synoviocytes].

This paper aimed to investigate the effects of Jinwu Jiangu recipe total extract on the IL-17/STAT3 signals in rheumatoid arthritis synovial fibroblasts(RASF). The primary RASFs were cultured by tissue piece method in vitro， and divided into blank control group， Jinwu Jiangu recipe low dose group， Jinwu Jiangu recipe middle dose group， Jinwu Jiangu recipe high dose group， and tripterygium glycosides control group. They were then treated with corresponding serum free medium， different doses of Jinwu Jiangu recipe total extract(0.06， 0.6， 6.0 g·L⁻¹)， and tripterygium glycosides(0.03 g·L⁻¹) respectively for 24 hours. The gene expression levels of RORα， RORγt， and STAT3 mRNA were detected by polymerase chain reaction(PCR)， and the protein activity of IL-17R and pSTAT3 were measured by Western blot assay. The results showed that as compared with blank control group， the expression levels of RORα， RORγt， IL-17R and STAT3 mRNA in RASF were significantly declined(P<0.01). As compared with tripterygium glycosides control group， Jinwu Jiangu recipe total extract middle dose group and high dose group can down-regulate the expression levels of RORα， RORγt， IL-17R and STAT3 mRNA(P<0.05)， and the effect was more obvious in high dose group(P<0.01). As compared with blank control group， the protein expression levels of IL-17R and pSTAT3 in each treatment group were obviously decreased(P<0.01). As compared with tripterygium glycosides control group， Jinwu Jiangu recipe high dose group had more obvious effect in down-regulating the protein expression of pSTAT3(P<0.01). Therefore， Miao medicine Jinwu Jiangu recipe total extract can down-regulate the expressions of RORα， RORγt， and STAT3 mRNA， and inhibit the protein activity of IL-17R and pSTAT3 in RASF.

[Effect of Jinwu Jiangu Recipe on Expressions of NF-kappaB and IL-17 in Collagen Induced Arthritis Model Rats].

OBJECTIVE: To explore the effect of Jinwu Jiangu Recipe (JJR) on the expression of synovial cells' nuclear factor-kappaB (NF-kappaB) and serum interleukin 17 (IL-17) in collagen induced arthritis (CIA) rats. METHODS: Totally 60 Wistar rats were randomly divided into 6 groups, i.e., the blank control group, the model group, high, middle, and low dose JJR treatment groups, and the tripterygium control group, 10 in each group. Except rats in the blank control group, CIA model was established in rats of the rest 5 groups. Then they were treated from the 7th day of modeling. After 4 weeks of medication they were sacrificed, serum collected, and synovium of joints were isolated. The expression of serum IL-17 was detected in synovium of joints by enzyme linked immunosorbent assay (ELISA). And the expression of NF-kappaB/P65, Ikappabetaalpha and NF-KappaB/P50 were detected by Western blot. RESULTS: Compared with the blank control group, the serum IL-17 level increased in the model group (P <0. 01). Compared with the model group, the serum IL-17 level obviously decreased in high and middle dose JJR groups and the tripterygium control group (P < 0.01). Results of Western blot showed, when compared with the blank control group, protein activities of NF-kappaB/P65 and NF-kappaB/P50 were significantly enhanced in the model group (P < 0.01). Compared with the model group, protein activities of NF-kappaB/P65 and NF-kappaB/P50 significantly decreased in high and middle dose JJR groups and the tripterygium control group (P < 0.05, P < 0.01). All indices mentioned above were higher in the low dose JJR group than in the tripterygium control group (P < 0.05, P < 0.01). CONCLUSION: JJR could lower the expression of serum IL-17 in CIA model rats, and inhibit protein activities of NF-kappaB/P65 and NF-kappaB/P50.

[Chinese medical syndromes of rheumatoid arthritis: a clinical literature study].

OBJECTIVE: To explore the distribution features of Chinese medical syndromes of rheumatoid arthritis (RA) by literature retrieval. METHODS: Pertinent articles on treating RA by syndrome differentiation were retrieved from China National Knowledge Infrastructure Databases (CNKI), VIP Chinese Biomedical Journal Database, Guizhou Digital Library, and Duxiu Chinese Academic Periodicals full papers database from January 2000 to December 2011. RESULTS: A total of 33 documents were recruited covering 4 233 cases. Damp-heat blocking collaterals syndrome occupied the top in the occurrence frequency (20 times, 60.61%), followed by deficiency of Gan and Shen syndrome (18 times, 54.55%), intermingled phlegm and blood-stasis syndrome (17 times, 51.52%), wind-cold-damp impediment syndrome (15 times, 45.45%), cold-damp blocking collaterals syndrome (14 times, 42.42%), wind-damp-heat impediment and deficiency of qi and blood syndrome (10 times, 30.30%), and intermingled cold and heat syndrome (9 times, 27.27%). According to the case number of patients, it was sequenced as damp-heat blocking collaterals syndrome syndrome (768 cases, 18.14%), wind-damp-heat impediment syndrome(666 cases, 15.73%), wind-cold-damp impediment syndrome(584 cases, 13.80%), cold-damp blocking collaterals syndrome syndrome (517 cases, 12.21%), intermingled cold and heat syndrome (415 cases, 9.80%), intermingled phlegm and blood-stasis syndrome (364 cases, 8.60%), deficiency of Gan and Shen syndrome (235 cases, 5.55%),asthenia of healthy energy due to lingering arthralgia syndrome (223 cases, 5.27%). The case numbers of remaining syndromes did not exceed 5%. CONCLUSION: Damp-heat blocking collaterals syndrome was the main syndrome in RA patients, followed by wind-cold-damp impediment syndrome,wind-damp-heat impediment syndrome,cold-damp blocking collaterals syndrome,intermingled phlegm and blood-stasis syndrome, and deficiency of Gan and Shen syndrome.

[Effect of jiedu huayu recipe containing serum on the expressions of survivin and NF-kappaB of K562/A02 cell apoptosis by].

OBJECTIVE: To study the effect of Jiedu Huayu Recipe containing serum on the expressions of survivin gene of leukemia K562/A02 cell apoptosis and the expression of nuclear factor kappa B (NF-kappaB). METHODS: K562/A02 cells were divided into six groups. An equal volume of calf serum, rabbit serum, high, middle, and low dose Jiedu Huayu Recipe containing serum, and interferon was respectively added. K562 sensitive cell strain was set up as the control. The expressions of survivin in K562/A02 cells after treated with Jiedu Huayu Recipe containing serum were detected using semi-quantitative reverse transcriptase polymerase chain reaction method. The expressions of NF-kappaB/P65, NF-kappaB/P50, and IKBa were detected using Westem blot method. RESULTS: The survivin resistant gene was highly expressed in K562/A02 cells. After treated by Jiedu Huayu Recipe containing serum, the expressions of survivin could be obviously lowered in the high and middle dose Jiedu Huayu Recipe containing serum groups as well as the interferon control group. NF-kappaB was also highly expressed in K562/A02 resistant cells. After treated by Jiedu Huayu Recipe containing serum, the expressions of P65, P50, and IkappaBalpha decreased to some degrees. The decrement was the most obvious in the middle and high dose Jiedu Huayu Recipe containing serum groups. The decreased expressions of P65 and P50 in the high dose Jiedu Huayu Recipe containing serum group were higher than that in the interferon control group (P<0.05). The decrement of IkappaBalpha was equivalent to that of the interferon control group (P>0.05). CONCLUSION: Jiedu Huayu Recipe could block the NF-kappaB signal pathway of K562/A02 cells and reverse drug resistance by influencing the expression of NF-kappaB-survivin genes.