Polysaccharides of Scrophularia ningpoensis Hemsl.: Extraction, Antioxidant, and Anti-Inflammatory Evaluation

Jian'an Wang, Lufen Huang, Qiang Ren, Yanjun Wang, Lirun Zhou, Yingjie Fu, Chunmei Sai, Shafii Shaibu Pella, Yingying Guo, Li-Na Gao, Jian'an Wang, Lufen Huang, Qiang Ren, Yanjun Wang, Lirun Zhou, Yingjie Fu, Chunmei Sai, Shafii Shaibu Pella, Yingying Guo, Li-Na Gao

Abstract

The roots of Scrophularia ningpoensis Hemsl. are a famous traditional Chinese medicinal herb and are also used as health food. However, information about polysaccharides from S. ningpoensis (SNPS) is very limited. We applied the ultrasonic-assisted extraction (UAE) process to extract SNPS. The UAE conditions were optimized using single-factor experiments and response surface analysis. Under the optimized conditions of ultrasonic power of 550 W, extraction time of 26 min, and extraction temperature at 50°C, the highest yield of 13.47% ± 1.63% was obtained, which was in accordance with the predicted value of 13.71%. In comparison with traditional hot water extraction, the optimized UAE method significantly increased the extraction yield with lower extraction temperature and shorter extraction time. Furthermore, the in vitro antioxidant evaluation showed that EC50 values of SNPS were 2.43 ± 0.21, 4.40 ± 0.35, and 0.56 ± 0.062 mg/mL for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) radical, hydroxyl free radical, and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay, respectively. The anti-inflammatory potential of SNPS was detected in lipopolysaccharide (LPS) induced ICR mice. Real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay showed that SNPS significantly improved LPS-stimulated inflammatory response by decreasing mRNA and protein expression of interleukin (IL)-6 and tumour necrosis factor (TNF)-α in a dose-dependent manner. In conclusion, the extraction process of SNPS established in this study is reliable, and SNPS possesses potential antioxidant and anti-inflammatory activities, which will provide a theoretical basis for guiding the clinical application of S. ningpoensis.

Conflict of interest statement

The authors declare that they have no competing interests.

Copyright © 2020 Jian'an Wang et al.

Figures

Figure 1
Figure 1
Effect of different extraction parameters on the yield of SNPS. (a) Ultrasonic power (W). (b) Extraction temperature (°C). (c) Extraction time (min).
Figure 2
Figure 2
Diagnostic plots for the model adequacy. (a) The plot of internally studentized residuals vs. the actual run. (b) Normal probability plot of the studentized residuals. (c) Residuals vs. predicted plot graph.
Figure 3
Figure 3
Interactive effects of ultrasonic power, ultrasonic temperature, and ultrasonic time on the yield of SNPS. (a and b) Response surface and contour plots of the effect of ultrasonic power and temperature on the extraction rate of SNPS; (c and d) Response surface and contour plots of the effect of ultrasonic time and temperature on the extraction rate of SNPS; (e and f) Response surface and contour plots of the effect of ultrasonic power and time on the extraction rate of SNPS.
Figure 4
Figure 4
Characterization of SNPS. (a) HPLC analysis of monosaccharide components of SNPS (1. Fucose; 2. Rhamnose; 3. Arabinose; 4. Galactosamine; 5. Galactose; 6. Glucose; 7. Xylose; 8. Mannose; 9. Galacturonic acid; 10. Glucuronic acid). (b) FT-IR analysis of SNPS.
Figure 5
Figure 5
Antioxidant activities of SNPS. (a) Scavenging of DPPH radical. (b) Scavenging of OH free radical. (c) Scavenging of ABTS radical. Data are presented as the mean means ± S.D., n = 4. Vc was used as a positive control.
Figure 6
Figure 6
Anti-inflammatory activities of SNPS on LPS-induced mice. Mice were treated with SNPS (100, 200, and 400 mg/kg), aspirin (10 mg/kg), or saline for 4 days (once per day) and stimulated with LPS (1 mg/kg) for 90 min. The protein expression of (a) IL-6 and (b) in serum were determined by ELISA. Values are means ± S.E. (n = 6). The mRNA expression of IL-6 (c) and TNF-α (d) in the liver was measured using real-time RT-PCR. Values are means ± S.E. ((n) = 4). The β-actin was used as an internal control for real-time RT-PCR. Aspirin was used as a positive control. The significant difference compared with the normal control group, ####P < 0.0001. The significant difference compared with LPS-treated alone, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001.

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