Isolation and purification of polysaccharides from Bupleurum marginatum Wall.ex DC and their anti-liver fibrosis activities

Abstract

Bupleurum marginatum Wall.ex DC [Apiaceae] (BM)is widely grown in southwestern China, and the whole plant is used as Traditional Chinese Medicine (TCM). Polysaccharides are main natural products in lots of TCM and have been studied for their effects of reducing oxidative stress, anti-inflammation and immune regulation. Herein, we investigated the extraction techniques of Bupleurum marginatum Wall.ex DC polysaccharides (BMP), the identification of their key components, and their ability to inhibit liver fibrosis in both cellular and animal models. Component identification indicated that monosaccharides in BMP mainly consisted of glucose, galactose, mannose, rhamnose, arabinose, and xylose. In vivo analysis revealed that BMP provided significant protective effects on N-Nitroso dimethylamine (NDMA)-induced liver fibrosis rats through reducing hepatomegaly, reducing tissue inflammation, and reducing collagen deposition. BMP also improved the hepatobiliary system and liver metabolism in accord to reduce the serum levels of ALT, AST, ALP, r-GT, and TBIL. In addition, BMP could reduce the level of inflammation and fibrosis through inhibition of IL-1β and TGF-β1. Cellular studies showed that the BMP could provide therapeutic effects on lipopolysaccharide (LPS)-induced cellular fibrosis model, and could reduce the level of inflammation and fibrosis by decreasing the level of TGF-β1, IL-1β, and TNF-α. Our study demonstrated that BMP may provide a new therapy strategy of liver injury and liver fibrosis.

Keywords: Bupleurum marginatum Wall.ex DC; activity study; anti-liver fibrosis; polysaccharides; water extraction and alcohol precipitation.

FIGURE 1

(A) DEAE-52 column elution curve. (B) UV scanning spectrum of the BMP-D.

FIGURE 2

(A) Ion chromatogram of mixed standard sample. (B) Ion chromatogram of BMP-D sample. (C) FT-IR scanning spectrum of BMP-D sample.

FIGURE 3

(A) Changes in body weight of rats over time. (B) Sirius red stained section positive area. ^## p < 0.01, versus control group. **p < 0.01, versus model group.

FIGURE 4

Effects of BMP on the histopathological changes of rat liver tissue. H&E staining (20.0×). (A) Control group, (B) Model group, (C) PC group, (D) L-BMP group, (E) M-BMP group, (F) H-BMP group.

FIGURE 5

Effects of BMP on the histopathological changes of rat liver tissue. Sirius red staining (10.0×). (A) Control group, (B) Model group, (C) PC group, (D) L-BMP group, (E) M-BMP group, (F) H-BMP group.

FIGURE 6

Effects of BMP on serum biochemical indexes in rats. (A) AST, (B) ALT, (C) ALP, (D) r-GT, (E) TBIL, (F) GLU. ^# p < 0.05, ^## p < 0.01, versus control group. *p < 0.05, **p < 0.01, versus model group.

FIGURE 7

Effect of BMP on biochemical indexes of rat liver tissue (A) TGF-β1, (B) IL-1β, (C) TNF-α, (D) α-SMA. ^# p < 0.05, ^## p < 0.01, versus control group. *p < 0.05, **p < 0.01, versus model group.

FIGURE 8

(A) Effects of different concentrations of BMP on the survival rate of BRL-3A cells. (B) Inhibitory effect of BMP on LPS-activated HSCs-T6 proliferation. Micrographs of HSCs-T6 (C) static, (D) LPS activation. ^# p < 0.05, ^## p < 0.01, versus control group. *p < 0.05, **p < 0.01, versus model group.

FIGURE 9

Effects of BMP on biochemical indexes of LPS-activated HSCs-T6. (A) TGF-β1, (B) TNF-α, (C) IL-1β, (D) α-SMA. ^## p < 0.01, versus control group. *p < 0.05, **p < 0.01, versus model group.