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. 2013:2013:580673.
doi: 10.1155/2013/580673. Epub 2013 Aug 22.

Matrine Inhibits Infiltration of the Inflammatory Gr1(hi) Monocyte Subset in Injured Mouse Liver through Inhibition of Monocyte Chemoattractant Protein-1

Duo Shi  1 , Jinjin Zhang, Lei Qiu, Jianzhong Li, Zhenlin Hu, Junping Zhang
Affiliations

Matrine Inhibits Infiltration of the Inflammatory Gr1(hi) Monocyte Subset in Injured Mouse Liver through Inhibition of Monocyte Chemoattractant Protein-1

Duo Shi et al. Evid Based Complement Alternat Med. 2013.

Abstract

Matrine (Mat) is a major alkaloid extracted from Sophora flavescens Ait, an herb which is used in the traditional Chinese medicine for treatment of inflammation, cancer, and other diseases. The present study examined the impact of Mat on the CCl4-induced hepatic infiltration of Gr1(hi) monocytes to explore the possible mechanisms underlying its anti-inflammatory and antifibrotic effects. The results indicated that Mat protected mice from acute liver injury induced by single intraperitoneal injection of CCl4 and attenuated liver fibrosis induced by repeated CCl4 injection. Meanwhile, the infiltrations of Gr1(hi) monocytes in both acute and chronic injured livers were all inhibited, and the enhanced hepatic expression of MCP-1 was suppressed. Cellular experiments demonstrated that Mat directly inhibited MCP-1 production in both nonparenchymal cells and hepatic stellate cells derived from CCl4-injured livers. Transwell chemotaxis assays showed that Mat significantly inhibited the chemotactic activity of MCP-1. These results suggest that the anti-inflammatory and antifibrotic effects of Mat could be contributed, at least in part, to its prevention of Gr1(hi) monocyte infiltration into the injured livers and inhibition of MCP-1 production and activity. These findings extend our understanding of the mechanisms underlying the anti-inflammatory and antifibrotic effects of Mat.

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Figures

Figure 1
Figure 1
Mat protected mice against CCl4-induced acute liver injury and reduced hepatic inflammatory infiltration. (a) C57BL/6 mice were injected i.p. with CCl4 (0.6 mL/kg) diluted in corn oil, and sera were collected and analyzed for ALT levels at indicated time points. Data are expressed as the mean ± (SD) (n = 8 mice/group). *P < 0.05 versus corn oil control. (b) Liver tissues were collected at indicated time points after CCl4 injection, and liver sections were made and stained with hematoxylin-eosin (HE) (original magnification ×200). (c) Mat (10 mg/kg, 30 mg/kg) was administrated orally 3 h before CCl4 injection, and sera were collected and analyzed for ALT levels at 24 h after CCl4 injection. Data are expressed as the mean ± (SD) (n = 8 mice/group). # P < 0.05 versus normal group; *P < 0.05 versus CCl4-alone group. (d) Liver tissues were collected 24 h after CCl4 injection, and liver sections were made and stained with HE (original magnification ×200).
Figure 2
Figure 2
Gr1hi but not Gr1lo monocytes were massively recruited into CCl4-injured livers. (a) Flow cytometric analysis of intrahepatic monocytes. Non-parenchymal cells (NPCs) were isolated from CCl4-injured livers, and leukocytes were firstly gated on CD45+; then two monocyte subsets were distinguished based on CD11b and F4/80 expressions. The CD11b+F4/80+ subset (right upper gate) expresses high Gr1; the CD11b+F4/80− population expresses low Gr1. The percentage of cells in selected gates is indicated. (b) NPCs were isolated from livers of different experimental groups at 24 h and 48 h after CCl4 injection. The total number of NPCs, the percentage and absolute number of CD45+ leukocytes in NPCs, and the percentage and absolute number of different monocyte subsets in NPCs were analyzed by flow cytometry using the strategy depicted in (a). Data are expressed as the mean ± (SD) (n = 8 mice/group). *P < 0.05 versus normal control (0 h).
Figure 3
Figure 3
Mat inhibited the recruitment of Gr1hi monocytes into acute liver injury. C57BL/6 mice were injected i.p. with CCl4 (0.6 mL/kg). Mat (30 mg/kg) was administrated orally 3 h before CCl4 injection. NPCs were isolated from livers of different experimental groups at 24 after CCl4 injection. (a) The total number of NPCs, (b) the percentage of CD45+ leukocytes in NPCs, (c) the absolute number of CD45+ leukocytes in NPCs, (d) the percentage of different monocyte subsets in NPCs, and (e) the absolute number of different monocyte subsets were analyzed by flow cytometry. Data are expressed as the mean ± (SD) (n = 8 mice/group). *P < 0.05 versus CCl4-only group; # P < 0.05 versus normal control group.
Figure 4
Figure 4
Administration of Mat reduced MCP-1 production in CCl4-challenge mice. C57BL/6 mice were injected i.p. with CCl4 (0.6 mL/kg). Mat (30 mg/kg) was administrated orally 3 h before CCl4 injection. (a) Sera were collected and analyzed for MCP-1 levels 24 h after CCl4 injection. (b) Liver tissues were collected 24 h after CCl4 injection and analyzed for MCP-1 mRNA levels with real-time PCR. Data in (a) and (b) are expressed as the mean ± (SD) (n = 8 mice/group). *P < 0.05 versus CCl4-only group; # P < 0.05 versus normal control group. (c) Liver tissues were collected 24 h after CCl4 injection; paraffin sections were made and analyzed for MCP-1 with immunohistochemistry (original magnification ×200).
Figure 5
Figure 5
Mat inhibited MCP-1 production in NPCs and HSCs. (a) NPCs were isolated from CCl4-injured livers and stimulated with LPS (1 μg/mL) for 24 h in the absence or presence of the indicated concentrations of Mat. MCP-1 levels in the supernatants were measured by ELISA. (b) HSCs were isolated from CCl4-injured livers and stimulated with TNF-α (30 ng/mL) for 24 h in the absence or presence of Mat. MCP-1 mRNA levels were measured by real-time PCR. Data were obtained from 3 independent experiments and presented as mean ± (SD). *P < 0.05 versus LPS or TNF-α alone;  # P < 0.05 versus nontreatment control group.
Figure 6
Figure 6
Mat inhibited the chemotactic activity of MCP-1. Chemotaxis assay was carried out with transwell culture chambers. Culture mediums containing 10 ng/mL MCP-1 were added to the lower wells of the chambers, and 1 × 105 murine PBMCs were seeded into the upper wells in the absence or presence of the indicated concentrations of Mat. (a) After 2 h incubation, the cells migrating to the lower surface of the membrane were examined under microscope after being fixed with methanol and stained with crystal violet (original magnification ×100). (b) Four different areas of migrated cells were counted for each data point, and the number of migrated cells per well was calculated. Data were obtained from 3 independent experiments and presented as mean ± (SD).  *P < 0.05 versus MCP-1 alone;  # P < 0.05 versus nontreatment control group.
Figure 7
Figure 7
Mat exhibited antifibrotic effects and inhibited hepatic infiltration of Gr1hi monocytes and production of MCP-1. C57BL/6 mice were injected i.p. with CCl4 (0.6 mL/kg, diluted in corn oil) or the same volume of corn oil twice weekly for 6 weeks. Mat (10 mg/kg) was orally administrated 5 times weekly for the last 3 weeks. Mice were sacrificed for analysis at 48 h after the last CCl4 injection. (a) Liver sections were stained with H&E, Sirius red, and Masson trichrome, respectively (original magnification ×100). (b) Liver hydroxyproline content was measured. (c) NPCs were isolated from livers, and the total number of NPCs, the percentage of CD45+ leukocytes in NPCs, the absolute number of CD45+ leukocytes, the percentage of different monocyte subsets in NPCs, and the absolute number of different monocyte subsets were analyzed with flow cytometry using the strategy as depicted in Figure 2. (d) Hepatic MCP-1 mRNA levels were analyzed with real-time PCR. Data in (b)–(d) are expressed as the mean ± (SD) (n = 8 mice/group).  *P < 0.05 versus CCl4-only group; # P < 0.05 versus normal control group.
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References

    1. Friedman SL. Liver fibrosis—from bench to bedside. Journal of Hepatology, Supplement. 2003;38(1):S38–S53. - PubMed
    1. Friedman SL. Mechanisms of hepatic fibrogenesis. Gastroenterology. 2008;134(6):1655–1669. - PMC - PubMed
    1. Bataller R, Brenner DA. Liver fibrosis. Journal of Clinical Investigation. 2005;115(2):209–218. - PMC - PubMed
    1. Winwood PJ, Arthur MJP. Kupffer cells: Their activation and role in animal models of liver injury and human liver disease. Seminars in Liver Disease. 1993;13(1):50–59. - PubMed
    1. Shiratori Y, Geerts A, Ichida T. Kupffer cells from CCl4-induced fibrotic livers stimulate proliferation of fat-storing cells. Journal of Hepatology. 1986;3(3):294–303. - PubMed