Silencing of CD147 inhibits hepatic stellate cells activation related to suppressing aerobic glycolysis via hedgehog signaling

Abstract

Hepatic stellate cells (HSCs) activation is a key step that promotes hepatic fibrosis. Emerging evidence suggests that aerobic glycolysis is one of its important metabolic characteristics. Our previous study has reported that CD147, a glycosylated transmembrane protein, contributes significantly to the activation of HSCs. However, whether and how it is involved in the aerobic glycolysis of HSCs activation is unknown. The objective of the present study was to validate the effect of CD147 in HSCs activation and the underlying molecular mechanism. Our results showed that the silencing of CD147 decreased the expression of α-smooth muscle-actin (α-SMA) and collagen I at both mRNA and protein levels. Furthermore, CD147 silencing decreased the glucose uptake, lactate production in HSCs, and repressed the lactate dehydrogenase (LDH) activity, the expression of hexokinase 2 (HK2), glucose transporter 1 (Glut1). The effect of galloflavin, a well-defined glycolysis inhibitor, was similar to CD147 siRNA. Mechanistically, CD147 silencing suppressed glycolysis-associated HSCs activation through inhibiting the hedgehog signaling. Moreover, the hedgehog signaling agonist SAG could rescue the above effect of CD147 silencing. In conclusion, CD147 silencing blockade of aerobic glycolysis via suppression of hedgehog signaling inhibited HSCs activation, suggesting CD147 as a novel therapeutic target for hepatic fibrosis.

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Acknowledgements

This work was supported by grants from the National Natural Science Fund of China (Grant No. 81700546),Scientific Research Project of Xi’an Medical University (Grant Nos. 2017PT13; 2018PT68), Key Research Project in Shaanxi Province (Grant Nos. 2018SF-168; 2020SF-349), Natural Science Basic Research Program of Shaanxi (Grant No. 2019JQ-886), National Science and Technology Fund research Program of Xi’an Medical University (Grant No. 2017GFY18).

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Correspondence to Haiyan Li.

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Supplemental Fig. 1. CD147 is overexpressed in activated HSCs, and CD147 silencing inhibits the HSCs activation. Measurement of CD147, α-SMA, collagen protein levels (A) in HSC-T6 cells stimulated with various doses of TGF-β1 by western blots. The protein levels of CD147 knocked down by 2 different siRNAs were measured in LX-2 (B) and HSC-T6 (C) cells by western blots. Measurement protein levels of CD147 (D) in si-CD147 treated HSC-T6 cells exposed to 4ng/ml TGF-β1Supplemental Fig. 1. CD147 is overexpressed in activated HSCs, and CD147 silencing inhibits the HSCs activation. Measurement of CD147, α-SMA, collagen protein levels (A) in HSC-T6 cells stimulated with various doses of TGF-β1 by western blots. The protein levels of CD147 knocked down by 2 different siRNAs were measured in LX-2 (B) and HSC-T6 (C) cells by western blots. Measurement protein levels of CD147 (D) in si-CD147 treated HSC-T6 cells exposed to 4ng/ml TGF-β1 (TIF 887 kb)

Supplemental Fig. 2. CD147 silencing inhibits the HSCs activation through suppressing aerobic glycolysis. TGF-β1, si-CD147, and/or galloflavin(a lactate dehydrogenase inhibitor) were applied to HSC-T6 cells at indicated concentrations for 24 h. A. Measurement of α-SMA, collagen I protein levels by western blots. B. Measurement of HK2, Glut1 protein levels by western blots (TIF 258 kb)

Supplemental Fig. 3. Silencing of CD147 inhibits Hedgehog signaling in activated HSCs. TGF-β1,si-CD147, and/or SAG (a Hedgehog signaling activator) were applied to HSC-T6 cells at indicated concentrations for 24 h. Western blots analyses of protein levels of Patched, Smo, and Hhip (TIF 563 kb)

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Li, H., Yang, L., Sun, Y. et al. Silencing of CD147 inhibits hepatic stellate cells activation related to suppressing aerobic glycolysis via hedgehog signaling. Cytotechnology (2021). https://doi.org/10.1007/s10616-021-00460-9

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Keywords

  • CD147
  • Hepatic stellate cells(HSCs)
  • Aerobic glycolysis
  • Hepatic fibrosis