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Journal of Peking University (Health Sciences) ›› 2025, Vol. 57 ›› Issue (5): 926-933. doi: 10.19723/j.issn.1671-167X.2025.05.017

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Effect of the combination of alkaloids from Euodiae Fructus and berberine in Zuojin Pill on cytotoxicity in HepG2 cells

Yadong GAO1,2, An ZHU1,3, Ludi LI1, Yingzi LI1, Qi WANG1,4,5,*()   

  1. 1. Department of Toxicology, Peking University School of Public Health, Beijing 100191, China
    2. Fujian Provincial Key Laboratory of Zoonosis Research, Fujian Provincial Center for Disease Control and Prevention, Fuzhou 350012, China
    3. Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350108, China
    4. Key Laboratory of State Administration of Traditional Chinese Medicine for Compatibility Toxicology, Beijing 100191, China
    5. Beijing Key Laboratory of Toxicological Research and Risk Assessment for Food Safety, Beijing 100191, China
  • Received:2024-02-07 Online:2025-10-18 Published:2025-05-07
  • Contact: Qi WANG
  • Supported by:
    the National Key Research and Development Program of China(2018YFC1704500); the National Key Research and Development Program of China(2018YFC1704506)

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Abstract: Objective: To investigate the hepatotoxicity of alkaloids from Euodiae Fructus combined with berberine (BBR) in Zuojin Pill, and to preliminarily explore the possible detoxification mechanism of the combination components. Methods: The combination ratio of components was determined by the maximum concentration (Cmax) of the chemical components in Zuojin Pill. HepG2 cell model was used to investigate the combined toxicity of the hepatotoxic components from Euodiae Fructus, such as evodiamine (EVO) or dehydroevodiamine (DHED), with BBR for 48 h. The experimental groups were set as follows: the vehicle control group, the EVO group, the DHED group, the BBR group, and the combination group of EVO or DHED with BBR. The cell counting kit-8 (CCK-8) method was used to determine the cell viability, and the combination index (CI) was used to determine the combined toxicity of the components. The alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydroge-nase (LDH), and alkaline phosphatase (ALP) activities as well as total bilirubin (TBIL) content in the cell culture supernatant were detected. The protein expression levels of bile acid transporters, such as bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP2), were detected by Western blot. The intracellular malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in HepG2 cells were detected. Results: Compared with EVO or DHED group, the combination of EVO 1 μmol/L with BBR 10 μmol/L or DHED 50 μmol/L with BBR 35 μmol/L significantly increased cell viability of HepG2 cells (P < 0.01), with CI values of 77.89 or 4.49, respectively, much greater than 1. Significant decreases in the activities of ALT, AST, LDH, ALP, and TBIL content in the cell culture supernatant were found in both combination groups (P < 0.05, P < 0.01). Compared with the EVO group, the combination of EVO with BBR upregulated the protein expression levels of BSEP and MRP2. Compared with the DHED group, the combination of DHED with BBR significantly downregulated the protein expression levels of BSEP and MRP2 (P < 0.01). Compared with EVO or DHED group, the combination of EVO or DHED with BBR significantly reduced the MDA content in HepG2 cells (P < 0.05, P < 0.01). Conclusion: A certain ratio of BBR combined with EVO or DHED had an antagonistic effect on HepG2 cytotoxicity, which might be related to regulating the expression of bile acid transpor-ters, and reducing lipid peroxidation damage.

Key words: Evodiamine, Dehydroevodiamine, Berberine, Toxicity of Chinese medicine, Chemical and drug induced liver injury

CLC Number: 

  • R114

Figure 1

Effects of EVO, DHED and BBR treatment for 48 h on the cell viability of HepG2 cells ($\bar x \pm s$, n=3) A, the cells were treated with EVO 1 μmol/L, BBR 10 μmol/L, or EVO 1 μmol/L+BBR 10 μmol/L; B, the cells were treated with DHED 50 μmol/L, BBR 35 μmol/L, or DHED 50 μmol/L+BBR 35 μmol/L for 48 h, respectively. **P < 0.01, compared with the control group; ##P < 0.01, compared with the EVO or DHED group. EVO, evodiamine; BBR, berberine; DHED, dehydroevodiamine."

Figure 2

Effects of EVO, DHED and BBR treatment for 48 h on the liver function indicators of HepG2 cells ($\bar x \pm s$, n=3) Group A and group B are the same as Figure 1. * P < 0.05, ** P < 0.01, compared with the control group; # P < 0.05, ## P < 0.01, compared with the EVO or DHED group. EVO, evodiamine; BBR, berberine; DHED, dehydroevodiamine; ALT, alanine transaminase; AST, aspartate amino-transferase; LDH, lactate dehydrogenase; ALP, alkaline phosphatase; TBIL, total bilirubin."

Figure 3

Effects of EVO, DHED and BBR treatment for 48 h on the expression levels of BSEP and MRP2 of HepG2 cells ($\bar x \pm s$, n=3) The protein expression levels were detected by Western blot, and the semi-quantitative analysis was performed. Group A and group B are the same as Figure 1. * P < 0.05, ** P < 0.01, compared with the control group; ## P < 0.01, compared with the EVO or DHED group. EVO, evodiamine; BBR, berberine; DHED, dehydroevodiamine; BSEP, bile salt export pump; MRP2, multidrug resistance-associated protein 2."

Figure 4

Effects of EVO, DHED and BBR treatment for 48 h on MDA content and SOD activity of HepG2 cells ($\bar x \pm s$, n=3) Group A and group B are the same as Figure 1. * P < 0.05, ** P < 0.01, compared with the control group; # P < 0.05, ## P < 0.01, compared with the EVO or DHED group. EVO, evodiamine; BBR, berberine; DHED, dehydroevodiamine; MDA, malondialdehyde; SOD, superoxide dismutase."

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[1] GAO Ya-dong,ZHU An,LI Lu-di,ZHANG Tao,WANG Shuo,SHAN Dan-ping,LI Ying-zi,WANG Qi. Cytotoxicity and underlying mechanism of evodiamine in HepG2 cells [J]. Journal of Peking University (Health Sciences), 2021, 53(6): 1107-1114.
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