Effect of Yiqi Yangyin Zhuyu Recipe in Anti-inflammation Activities in Vitro on LPS-induced RAW 264.7 Cells
10.13422/j.cnki.syfjx.20190129
- VernacularTitle: 益气养阴逐瘀方对LPS诱导的RAW 264.7细胞炎症模型体外抗炎活性的影响
- Author:
Lei TANG
1
;
Jian WU
1
;
Gu-ran YU
1
;
Ji-hong CHU
1
;
Shi-hai YAN
1
;
Hai-dan WANG
1
;
Guo-liang DAI
1
;
Xu HAN
1
Author Information
1. Affiliated Hospital of Nanjing University of Traditional Chinese Medicine, Nanjing 210029, China
- Publication Type:Research Article
- Keywords:
atheroscberosis (AS);
Yiqi Yangyin Zhuyu recipe;
macrophage;
classically activated macrophages (M1);
alternatively activated macrophages (M2);
lipopolysaccharide (LPS);
Gualou Xiebai Baijiu Tang;
Danggui Buxue Tang
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(1):141-148
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of Yiqi Yangyin Zhuyu recipe on expressions of inflammatory factor and transformation of classically activated macrophages(M1)/alternatively activated macrophages (M2) inflammatory phenotype in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Method: Methyl-thiazdyl-tetrazolium(MTT) reduction assay was used to detect the effect of different concentrations of Yiqi Yangyin Zhuyu recipe on the proliferation of the cells. The release of nitric oxide was detected by the Griess method. Enzyme linked immunosorbent assay(ELISA) was used to detect the release of M1/M2 inflammatory cytokines in cell supernatant. The expressions of the pro-inflammatory factor genes of M1-macrophages and the anti-inflammatory factor genes of M2-macrophages were detected by Real-time PCR. The protein expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1,nitric oxide synthase(iNOS) were detected by Western blot. Result: Results of MTT showed that Yiqi Yangyin Zhuyu recipe with the concentration of 2.0 g·L-1 and below had no effect on the cell proliferation. Results of Griess indicated that compared with blank group, the release of nitric oxide of LPS-induced group was increased (P<0.01); compared with LPS-induced group, Yiqi Yangyin Zhuyu recipe group could reduce the release of NO (P<0.01). Results of ELISA showed that compared with blank group, the release of M1/M2 inflammatory cytokines of LPS-induced group were both up-regulated (P<0.01); compared with LPS-induced group, the secretion of M1-macrophages inflammatory cytokines was down-regulated in Yiqi Yangyin Zhuyu recipe group (P<0.01), but with no change in M2-macrophages. Results of Real-time PCR indicated that compared with blank group, the expressions of M1/M2 were both increased (P<0.01,P<0.05); compared with LPS-induced group, the expression levels of M1-macrophages were down-regulated in traditional Chinese medicine group (P<0.01); anti-inflammatory factor genes of M2-macrophages had no significant change. Results of Western blot showed that compared with blank group, expression levels of TNF-α,IL-6,IL-1β,iNOS were up-regulated (P<0.01); compared with LPS-induced group, the expressions of TNF-α,IL-6,IL-1β,iNOS were down-regulated in Yiqi Yangyin Zhuyu recipe group, especially at the concentration at 2.0 g·L-1 (P<0.01). Conclusion: Yiqi Yangyin Zhuyu recipe could effectively inhibit the inflammatory reaction induced by LPS. The anti-inflammatory mechanism of Yiqi Yangyin Zhuyu recipe may be related to inhibition of macrophages to M1 phenotype polarization, so as to play the role of regulating immune and reducing the release of inflammatory cytokines, like NO,TNF-α,IL-6,IL-1β.
