Protective Effects of Two Safflower Derived Compounds, Kaempferol and Hydroxysafflor Yellow A, on Hyperglycaemic Stress-Induced Podocyte Apoptosis via Modulating of Macrophage M1/M2 Polarization.

OBJECTIVE: The primary initiating mechanism in diabetes nephropathy (DN) is hyperglycemia-induced inflammation in which macrophage and podocyte play important roles. The present research is aimed at exploring the effects of kaempferol (Ka) and hydroxysafflor yellow A (HSYA) on classically activated (M1)/alternatively activated (M2) macrophage polarization and podocyte apoptosis under hyperglycaemic conditions in vitro. METHODS: (1) RAW264.7 cells were treated with 11.1 mM glucose (NG), 33.3 mM glucose (HG), Ka 4-8 µM, and HSYA 100-200 µM separately. The expressions of inducible nitric oxide synthase (iNOS), tumor necrosis factor- (TNF-) α, mannose receptor (CD206), and arginase- (Arg-) 1 were quantified by Western blotting and real-time quantitative PCR. The collected supernatants from macrophage were named as (NG) MS, (HG) MS, (Ka) MS, and (HSYA) MS. (2) The podocyte survival rate was assessed by Bromodeoxyuridine assay, while TNF-α and interleukin- (IL-) 1ß levels were evaluated by Elisa. RESULTS: (1) Compared to the HG group, the Ka and HSYA 100 µM groups decreased iNOS and TNF-α levels and increased Arg-1 and CD206 expressions significantly (protein and mRNA: p < 0.05, respectively). (2) The podocyte survival rate of Ka 8 µM was higher than that of HG, and the rates of (Ka) MS and (HSYA 100 µM) MS were higher than that of (HG) MS significantly (all: p < 0.05). (3) TNF-α and IL-1ß levels of Ka and HSYA 100 µM were significantly lower than those of the HG group, and both levels in the (Ka) MS and (HSYA) MS were lower than those in the (HG) MS group significantly (p < 0.05, respectively). CONCLUSION: The protective effects of Ka and HSYA on podocyte apoptosis under hyperglycemic stress are related to their modulation on M1/M2 polarization and the lowering effects on TNF-α and IL-1ß levels.

Comparison of Inhibitory Effects of Safflower Decoction and Safflower Injection on Protein and mRNA Expressions of iNOS and IL-1ß in LPS-Activated RAW264.7 Cells.

OBJECTIVE: Safflower has antioxidant and anti-inflammatory activities. The two forms of preparations for safflower which are widely used in China are injection and decoction. The first step of the process for preparing an injection involves extracting safflower with water, which actually yields a decoction. This study is intended to investigate how the preparation process influences the anti-inflammatory activity of safflower in vitro. METHODS: Five samples, including a decoction (sample 1) and an injection (sample 5) of safflower, were prepared according to the national standard WS3-B-3825-98-2012 and were analyzed by the oxygen radical absorbance capacity (ORAC) method and the 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) method for comparison. Sample 1 and sample 5 were further tested by the Griess assay and ELISA for their effects on nitric oxide (NO) production and interleukin- (IL-) 1ß content in lipopolysaccharide- (LPS-) activated RAW264.7 cells. The protein and mRNA levels of inducible nitric oxide synthase (iNOS) and IL-1ß were measured by Western blotting and real-time quantitative PCR. RESULTS: Sample 5 showed a significantly higher ORAC value and a lower half inhibitory concentration (IC50) for DPPH scavenging activity as compared to the other four samples (p < 0.05). LPS significantly upregulated the mRNA and protein expressions of iNOS and IL-1ß as compared to the solvent control (p < 0.01). As compared to sample 1, sample 5 significantly decreased NO production, iNOS protein expression, and the contents of IL-1ß mRNA and IL-1ß protein at both 100 µg/ml and 200 µg/ml (all: p < 0.05) and significantly downregulated iNOS mRNA expression at 100 µg/ml (p < 0.05). CONCLUSIONS: Results of this study demonstrate that the safflower injection prepared according to the national standard has a significant effect of suppressing protein and mRNA expressions of iNOS and IL-1ß as compared to its traditional decoction.

Effects of dietary administration of Rose hip and Safflower on growth performance, haematological, biochemical parameters and innate immune response of Beluga, Huso huso (Linnaeus, 1758).

The aim of this study was to investigate effects of two dietary medicinal herbs, Rose hip (Rosa canina) and Safflower (Carthamus tinctorius) supplementation on growth performance, haematological, biochemical parameters and innate immune response of in juvenile beluga, Huso huso. Fish (26.3 ± 0.4 g) were allocated into 15 tanks (20 fish per tank) and triplicate groups were fed a control diet or diets containing 1% and 2% of medicinal herbs, respectively. Feed conversion ratio (FCR), specific growth rate (SGR) and condition factor (CF) did not show significant differences (P > 0.05) in fish given herbal diets. Significant differences were observed in number of white blood cells (WBC) and haemoglobin (Hb) values among the dietary treatments. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were significantly lower in supplemented diet groups compared with the control. Innate immune responses (lysozyme activity and ACH50) were significantly higher in 2% Safflower-fed fish compared with other groups (P < 0.05). These results indicate that medicinal herbs in diets can be considered as a beneficial dietary supplement for improving the physiological parameters and enhance the immune response of Persian sturgeon.