Effect of Cordyceps polysaccharides on hepatocyte apoptosis induced by tumor necrosis factor-α

Jianan ZHAO; Jinghua PENG; Ping LIU; Yiyang HU

doi:10.3969/j.issn.1001-5256.2021.06.029

Volume 37 Issue 6

Jun. 2021

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Article Navigation > Journal of Clinical Hepatology > 2021 > 37(6): 1368-1372

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Effect of Cordyceps polysaccharides on hepatocyte apoptosis induced by tumor necrosis factor-α

DOI: 10.3969/j.issn.1001-5256.2021.06.029

Jianan ZHAO¹,
Jinghua PENG¹,
Ping LIU^{1, 2, 3},
Yiyang HU^{1, 2, 3, 4
,
,}

1.
Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
2.
Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai 201203, China
3.
Key Laboratory of Liver and Kidney Diseases, Ministry of Education, Shanghai 201203, China
4.
Institute of Clinical Pharmacology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

Received Date: 2021-02-02
Accepted Date: 2021-03-10
Published Date: 2021-06-20

Abstract

Abstract

Objective To investigate the effect of Cordyceps polysaccharides on hepatocyte apoptosis and its mechanism of action. Methods Normal human L02 hepatocytes without any drug treatment was established as normal group. The L02 hepatocytes were treated with different concentrations of tumor necrosis factor-α (TNF-α) (5, 10, 20, and 40 ng/ml) for 24 hours to screen out the modeling conditions for the model of hepatocyte apoptosis, which were established as model group. According to the experimental design, L02 hepatocytes pretreated with three different concentrations of Cordyceps polysaccharides (50, 100, and 200 μg/ml) for 12 hours, with or without TNF-α treatment for 24 hours at the selected concentration, were established as experimental group. Related samples were collected for analysis. CCK8 assay was used to measure cell proliferation; Annexin V/PI double staining was used to measure the number of apoptotic hepatocytes; RT-PCR was used to measure the mRNA expression of cell apoptosis genes (Bax, caspase-3, caspase-8, caspase-9, and death receptor Fas); Western blotting was used to measure the protein expression of cleaved-caspase-3 and cleaved-caspase-8. A one-way analysis of variance or the Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and the least significant difference t-test or the Dunnett-t test was used for further comparison between two groups. Results CCK8 assay showed that compared with the normal group, the L02 hepatocytes induced by 40 ng/ml TNF-α for 24 hours had a significant reduction in proliferation (73.54%±14.19% vs 100.00%±23.61%, P < 0.01), and compared with the model group under the premise that Cordyceps polysaccharides had no obvious cytotoxicity to hepatocytes at the three concentrations, the 50, 100, and 200 μg/ml experimental groups had a significant increase in cell proliferation (108.10%±9.05%/114.30%±8.79%/117.70%±9.66% vs 93.02%±7.21%, all P < 0.01). Annexin V/PI double staining showed that compared with the normal group, the model group had a significant increase in the number of apoptotic cells (11.57%±1.41% vs 7.71%±1.20%, P < 0.05), and compared with the model group, the 50, 100, and 200 μg/ml experimental groups had a significant reduction in the number of apoptotic cells (15.16%±0.16%/13.28%±1.57%/16.91%±0.21% vs 18.91%±0.80%, all P < 0.05). RT-PCR and Western blotting showed that compared with the normal group, the L02 hepatocytes induced by 40 ng/ml TNF-α had significant increases in the mRNA expression of the apoptosis-related genes Bax, caspase-9, and death receptor Fas (all P < 0.05), as well as significant increases in the protein expression of the activated forms of cleaved-caspase-3 and cleaved-caspase-8 (P < 0.05). Compared with the model group, the 50 μg/ml experimental group had significant reductions in the mRNA expression of Bax, caspase-3, and caspase-9 and the protein expression of cleaved-caspase-8; the 100 μg/ml experimental group had significant reductions in the mRNA expression of Bax, caspase-3, caspase-8, Fas, and caspase-9 and the protein expression of cleaved-caspase-8; the 200 μg/ml experimental group had significant reductions in the mRNA expression of caspase-3, caspase-8, Fas, and caspase-9 and the protein expression of cleaved-caspase-3 and cleaved-caspase-8. The three concentrations of Cordyceps polysaccharides had a certain dose-effect trend. Conclusion Cordyceps polysaccharides can effectively inhibit the apoptosis of normal L02 hepatocytes induced by TNF-α.
- Liver Cirrhosis,
- Cordyceps Sinensis Mycelia Polysaccharide,
- Apoptosis

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Effect of Cordyceps polysaccharides on hepatocyte apoptosis induced by tumor necrosis factor-α

DOI: 10.3969/j.issn.1001-5256.2021.06.029

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