Mechanism of Si Junzitang in Inhibiting Hepatocellular Carcinoma Progression Through Regulating O-GlcNAcylation of NF-κB p65 in Paracancerous Tissues
10.13422/j.cnki.syfjx.20240129
- VernacularTitle:四君子汤调控肝癌癌旁组织NF-κB p65的O-GlcNAc糖基化修饰抑制肝细胞癌进展的机制
- Author:
Mingzhe LI
1
;
Zhaoqin FANG
1
;
Weifei ZHANG
2
;
Peike PENG
1
;
Dongwei JIA
1
Author Information
1. School of Traditional Chinese Medicine (TCM), Shanghai University of TCM, Shanghai 201203, China
2. School of Acupuncture-Moxibustion and Tuina, Shanghai University of TCM, Shanghai 201203, China
- Publication Type:Journal Article
- Keywords:
hepatocellular carcinoma;
Si Junzitang;
O-linked β-N-acetylglucosamine (O-GlcNAc);
nuclear factor kappa-B (NF-κB)
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(15):21-28
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the molecular mechanism by which Si Junzitang in intervening in the development of hepatocellular carcinoma (HCC) by regulating the O-linked β-N-acetylglucosamine modification (O-GlcNAcylation) of nuclear factor kappa-B (NF-κB) p65 in the paracancerous tissues. MethodThe orthotopic liver cancer mouse model was established. Twenty-four C57BL/6 mice were randomly divided into four groups: Normal group, model group, Si Junzitang low-dose group (10 g·kg-1), and Si Junzitang high-dose group (25 g·kg-1), with 6 mice in each group. The O-GlcNAcylation level and phosphorylation modification level of p65 in the paracancerous tissues were detected using Western blot. The O-GlcNAcylation of p65 was assessed using immunoprecipitation (IP). The mRNA expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor A (VEGFA), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) in the paracancerous tissues was detected using real-time quantitative polymerase chain reaction (Real-time PCR). The tumor number, liver weight, locomotor activity, grip strength, and Qi status of the mice were observed and analyzed. ResultCompared with the normal group, the model group showed a significant decrease in O-GlcNAcylation in the paracancerous tissues (P<0.01), a significant decrease in p65 O-GlcNAcylation (P<0.01), a significant increase in p65 phosphorylation (P<0.01), significantly elevated mRNA levels of cytokines IL-6, TNF-α, TGF-β1, VEGFA, MMP-2, and MMP-9 (P<0.01), significantly increased liver weight (P<0.01), significantly declined grip strength, number of grid crossings, and number of vertical stand-ups (P<0.01), and significantly dwindled Qi status (P<0.01). Compared with model group, the Si Junzitang low-dose and high-dose groups showed significantly increased levels of O-GlcNAcylation in the paracancerous tissues (P<0.05, P<0.01), significantly upregulated p65 O-GlcNAcylation levels (P<0.05, P<0.01), and significantly decreased p65 phosphorylation levels (P<0.01). In the Si Junzitang low-dose group, the mRNA levels of IL-6, TGF-β1, and VEGFA significantly decreased (P<0.05, P<0.01). In the Si Junzitang high-dose group, the mRNA levels of IL-6, TNF-α, TGF-β1, VEGFA, MMP-2, and MMP-9 significantly decreased (P<0.01), the number of tumors larger than 3 mm in diameter significantly decreased (P<0.01), and liver weight significantly decreased (P<0.05). Additionally, grip strength, number of grid crossings, and number of vertical stand-ups significantly increased (P<0.05, P<0.01), along with a significant increase in qi status (P<0.01). ConclusionSi Junzitang can inhibit the progression of orthotopic HCC in mice, which may be achieved by increasing the O-GlcNAcylation level in the paracancerous tissues, enhancing the O-GlcNAcylation of p65, inhibiting the phosphorylation modification of p65, and ultimately suppressing the expression of downstream IL-6, TNF-α, TGF-β1, VEGFA, MMP-2, and MMP-9.