ABSTRACT
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 ， 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(<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(<0.05)， and the effect was more obvious in high dose group(<0.01). As compared with blank control group， the protein expression levels of IL-17R and pSTAT3 in each treatment group were obviously decreased(<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(<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.
Subject(s)
Humans , Arthritis, Rheumatoid , Cells, Cultured , Drugs, Chinese Herbal , Pharmacology , Fibroblasts , Gene Expression Regulation , Nuclear Receptor Subfamily 1, Group F, Member 1 , Metabolism , Nuclear Receptor Subfamily 1, Group F, Member 3 , Metabolism , Receptors, Interleukin-17 , Metabolism , STAT3 Transcription Factor , Metabolism , Synovial Membrane , SynoviocytesABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>
Subject(s)
Animals , Rats , Arthritis, Experimental , Drug Therapy , Metabolism , Drugs, Chinese Herbal , Chemistry , Interleukin-17 , Blood , NF-kappa B , Metabolism , Random Allocation , Rats, Wistar , Synovial Membrane , Metabolism , Tripterygium , ChemistryABSTRACT
<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>