Mechanism of Ganshuang granule extract in alleviating N-acetyl-p-aminophenol-induced hepatocellular injury

Qiao WU; Pengfei YU; Yanzhen BI; Baozeng WANG; Zixuan WANG; Zhijie LI; Yu CHEN; Zhongping DUAN

doi:10.3969/j.issn.1001-5256.2021.01.024

Volume 37 Issue 1

Jan. 2021

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Article Navigation > Journal of Clinical Hepatology > 2021 > 37(1): 120-125

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Mechanism of Ganshuang granule extract in alleviating N-acetyl-p-aminophenol-induced hepatocellular injury

DOI: 10.3969/j.issn.1001-5256.2021.01.024

Qiao WU^{1, 2},
Pengfei YU¹,
Yanzhen BI¹,
Baozeng WANG²,
Zixuan WANG²,
Zhijie LI³,
Yu CHEN^{1
,
,},
Zhongping DUAN^{1
,
,}

1.
a. Intractable Hepatic Diseases and Artificial Liver Treatment & Training Center, b. Beijing Municipal Key Laboratory of Liver Failure and Artificial Liver Treatment Research, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China
2.
Center of Infectious Diseases, Beijing Tiantan Hospital, Capital Medical University, Beijing 100070, China
3.
Center of Hepatobiliary Surgery, The Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039, China

Received Date: 2020-06-05
Accepted Date: 2020-09-16
Published Date: 2021-01-20

Abstract

Abstract

Objective To investigate the ability of Ganshuang granule (a liver-protecting drug widely used in clinical practice) extract to reduce N-acetyl-p-aminophenol (APAP)-induced hepatotoxicity and possible mechanisms. Methods A total of five cell culture groups were set up in this experiment, i.e., normal control group, APAP injury group, and three injury protection groups treated with different concentrations of Ganshuang granule extract. Then 20 mmol/L APAP was added to the cell culture medium and incubated for 24 hours to establish an in vitro model of drug-induced liver injury, and the injury protection groups were treated with different concentrations of Ganshuang granule extract (0.2, 1, and 5 μg/ml) in advance for 8 hours of incubation before APAP were added for 24 hours. Related markers were measured, including the markers for hepatocellular injury [alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH)], the markers for mitochondrial injury [mitochondrial membrane potential, and glutamate dehydrogenase (GDH)], and antioxidant and oxidative stress markers [glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS)]. Related mechanism was discussed based on the experimental results. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Ganshuang granule extract alleviated APAP-induced hepatotoxicity, improved cell viability (P < 0.001), and reduced the levels of AST, ALT, and LDH in supernatant (P < 0.001, P < 0.001, and P < 0.05). Ganshuang granule extract inhibited APAP-induced hepatocellular oxidative stress, and compared with the APAP group, the Ganshuang granule extract groups had significant reductions in the oxidative stress indicators ROS and MDA (both P < 0.01). Ganshuang granule extract alleviated the loss of mitochondrial membrane potential induced by APAP (P < 0.05) and reduced the content of the mitochondrial injury marker GDH in supernatant (P < 0.001) in a dose-dependent manner. Ganshuang granule extract inhibited the expression of CYP2E1/1A2 (both P < 0.05) and increased the expression of phase Ⅱ enzymes in hepatocytes. Ganshuang granule extract induced the expression of Nrf2 and its downstream genes NQO-1 and GCLC (all P < 0.05). Conclusion Ganshuang granule extract can prevent APAP-induced hepatocellular injury through two ways. The first way is that Ganshuang granule extract downregulates the expression of CYP2E1/1A2 and thus reduces the production of NAPQI, a toxic product of APAP; the second way is that Ganshuang granule extract upregulates the expression of the detoxification pathway, which can activate Nrf2 to increase the expression of antioxidant enzymes (SOD and GSH) and phase Ⅱ enzymes and thus accelerate the harmless metabolism of APAP.
- Drug-Induced Liver Injury,
- Acetaminophen,
- Ganshuang Granules

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References

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Mechanism of Ganshuang granule extract in alleviating N-acetyl-p-aminophenol-induced hepatocellular injury

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