ABSTRACT
Objective:To investigate the effect of salvianolic acid C (SalC) on fibroblast-like synoviocytes and through the role of nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway.Methods:Rheumatoid arthritis-fibroblast-like synoviocytes (RA-FLSs) were exposed to different concentrations of SalC (0.1 μmol/L, 1 μmol/L, 5 μmol/L, 10 μmol/L, 20 μmol/L) for 24-72 h and measured for viability, proliferation, migration and invasion by Cell counting kit 8 (CCK-8) assay, wound-healing and transwell assay. The levels of Tumor Necrosis Factor-α (TNF-α), Interleukin-1 beta (IL-1β) and IL-6 were measured by enzyme linked immunosorbent assay (ELISA). Western blot was used to detect the expression of matrix metallopeptidase (MMP)-9, MMP-13, apoptosis-related proteins and Nrf2 mediated gene. Then we used ML385 to inhibit Nrf2 signaling pathway. RA-FLSs were measured for migration and invasion, and the expression of proteins related to apoptosis, inflammation and Nrf2 pathway.Results:Compared with the control group, SalC inhibited the cell migration significantly (0.1 μmol/L, 0.75±0.05, t=7.65, P<0.001; 5 μmol/L, 0.50±0.05, t=14.25, P<0.001; 10 μmol/L, 0.26±0.05, t=20.67, P<0.001) and invasion (0.1 μmol/L, 0.75±0.11, t=4.93, P<0.001; 5 μmol/L, 0.49±0.06, t=10.32, P<0.001; 10 μmol/L, 0.26±0.07 , t=14.96, P<0.001) of RA-FLs, reduced the levels of MMP-9 (0.1 μmol/L, 0.72±0.10, t=5.60, P<0.001; 5 μmol/L, 0.48±0.08, t=11.03, P<0.001; 10 μmol/L, 0.27±0.06, t=15.94, P<0.001) and MMP-13 (0.1 μmol/L, 0.77±0.06, t=8.66, P<0.001; 5 μmol/L, 0.58±0.06, t=11.03, P<0.001; 10 μmol/L, 0.32±0.13, t=14.22, P<0.001), and promoted apoptosis. SalC reduced the level of pro-inflammatory cytokines significantly ( P<0.001) and activated the expression of Nrf2 signaling pathway proteins Nrf2, CAT, NQO1, SOD1 and GSS ( P<0.001). After ML385 was used to interfere Nrf2, the levels of SalC on Nrf2 pathway proteins, such as Nrf2 (0.68±0.06, t=5.08, P<0.001), CAT (1.44±0.12, t=4.77, P<0.001), NQO1 (0.65±0.12, t=5.04, P<0.001), SOD1 (1.43±0.10, t=6.36, P<0.001) and GSS (1.42±0.10, t=7.60, P<0.001), as well as the levels of TNF-α [(260±22) pg/ml, t=13.75, P<0.001], IL-1β [(701±30) pg/ml, t=12.98, P<0.001], IL-6 [(180±10) pg/ml, t=16.38, P<0.001) were significantly reduced. In addition, ML385 inhibited the inhibition of SalC on cell migration and invasion (0.70±0.09, t=11.24, P<0.001; 0.64±0.04, t=8.03, P<0.001) and induction of apoptosis (24.4±1.8, t=23.02, P<0.001). Conclusion:SalC may inhibit cell activity and inflammatory response, promote apoptosis via the upregulation of Nrf2 signaling pathway. SalC may have therapeutic potential in RA. However, further investigation are needed in animal models and human.
ABSTRACT
Objective: To investigate the effect of water-soluble components from Salvia miltiorrhiza on liver sinusoid endothelial cell function and angiogenesis. Methods: Different dosages of water-soluble components from S. miltiorrhiza were incubated for 24 h with HHSEC cell line, and the toxicity of HHSEC cells was assayed by CCK-8 method. The proliferation of HHSEC cells was induced by endothelial cell growth supplements (ECGS), with receptor tyrosine kinase inhibitor sorafenib as positive control, cell proliferation was analyzed by EdU DNA cell proliferation kit; Fluorescence probe method was used to assay the intracellular NO level and NOS activity; Immunofluorescence method was performed to observe the expression of CD31; The transgenic zebrafishes were incubated for 24 h with each component. The fluorescence vessels, the number of functional blood vessels, and intersegmental vessel changes were observed; Chicken chorioallantoic membranes were incubated for 24 h with each component, Image Analysis Software was used to analyze the vessel area changes. Results: Compared with control group, the proliferation of HHSEC cells induced by ECGS increased, the level of NO and NOS activity reduced and the expression of CD31 increased; Compared with the model group, salvianolic acid A, salvianolic acid B, salvianolic acid C, and sorafenib inhibited the proliferation of HHSEC cells induced by ECGS, reduced the level of NO, NOS activity, and expression of CD31. The vessel area of chicken chorioallantoic membranes was decreased in sorafenib, salvianolic acid A, salvianolic acid B and caffeic acid. Salvianolic acid C also significantly inhibited the intersegmental vessel changes of transgenic zebrafish. Conclusion: Salvianolic acid A, salvianolic acid B, and salvianolic acid C have the potential effects on function of endothelial cell proliferation and angiogenesis.
ABSTRACT
OBJECTIVE: To study the water-soluble constituents of Clerodendranthus spicatus. METHODS: Various column chromatographic techniques were used to isolate the constituents, and their structures were identified by comparing their spectral data with those in literature. RESULTS: Fifteen compounds were isolated and elucidated as caffeic acid(1), caffeic acid methyl ester(2), caffeic acid ethyl ester(3), danshensu(4), ethyl 3, 4-dihydroxyphenyllactate(5), rosmarinic acid(6), methyl rosmarinate(7), ethyl rosmarinate(8), isorinic acid(9), dihydroconiferyl alcohol(10), dihydrosinapyl alcohol(11), salvianolic acid C(12), salvian-olic acid H(13), 3′-O-(8″-Z-caffeoyl) rosmarinic acid methyl ester(14) and baicalein(15). CONCLUSION: Compounds 2, 4-5 and 9-15 are obtained from this species for the first time.