十枣汤急性毒性及抗肿瘤疗效评价

doi:10.5246/jcps.2024.04.025

中国药学(英文版) ›› 2024, Vol. 33 ›› Issue (4): 329-338.DOI: 10.5246/jcps.2024.04.025

十枣汤急性毒性及抗肿瘤疗效评价

马立威¹^,², 陈哲³, 施国善⁴, 王玉静⁵, 陈颂⁵, 倪世宇⁶, 李京⁷, 葛鹏玲², 刘吉成¹^,^*()

^1. 齐齐哈尔医学院医药科学研究所(博士后工作站), 黑龙江齐齐哈尔 161006
^2. 黑龙江中医药大学博士后流动站, 黑龙江哈尔滨 150040
^3. 齐齐哈尔医学院公共卫生学院, 黑龙江齐齐哈尔 161006
^4. 贵州中医药大学基础医学院, 贵州贵阳 550025
^5. 齐齐哈尔医学院药学院, 黑龙江齐齐哈尔 161006
^6. 齐齐哈尔医学院附属第五医院, 黑龙江大庆 163001
^7. 齐齐哈尔医学院附属第三医院, 黑龙江齐齐哈尔 161099

收稿日期:2023-11-04 修回日期:2023-11-24 接受日期:2024-01-12 出版日期:2024-04-30 发布日期:2024-04-30
通讯作者: 刘吉成

Assessment of acute toxicity and antitumor efficacy of Shizao decoction

Liwei Ma¹^,², Zhe Chen³, Guoshan Shi⁴, Yujing Wang⁵, Song Chen⁵, Shiyu Ni⁶, Jing Li⁷, Pengling Ge², Jicheng Liu¹^,^*()

¹ Qiqihar Medical University, Medical Sciences Institution (Postdoctoral workstation) Qiqihar 161006, Heilongjiang, China
² Heilongjiang University of Chinese Medicine, Postdoctoral mobile station, Harbin 150040, Heilongjiang, China
³ Qiqihar Medical University, School of Public Health, Qiqihar 161006, Heilongjiang, China
⁴ Guizhou University of Traditional Chinese Medicine, School of Basic Medical Sciences, Guiyang 550025, Guizhou, China
⁵ Qiqihar Medical University, School of Pharmacy, Qiqihar 161006, Heilongjiang, China
⁶ The fifth affiliated hospital of Qiqihar Medical University, Daqing 163001, Heilongjiang, China
⁷ The third affiliated hospital of Qiqihaer Medical University, Qiqihar 161099, Heilongjiang, China

Received:2023-11-04 Revised:2023-11-24 Accepted:2024-01-12 Online:2024-04-30 Published:2024-04-30
Contact: Jicheng Liu
Supported by:
National Natural Science Foundation of China (Grant No. 82174207), the Science and Technology Project of Guizhou Province (Qiankehe Foundation-ZK [2021] General 502), the Growth Project of Young Scientific and Technological Talents in General Colleges and Universities in Guizhou Province (Qianjiaohe KY [2021] 208), and the Qiqihar Science and Technology Bureau Joint Guide Project (Grant No. LHYD-202028).

摘要/Abstract

摘要：

本研究旨在评价十枣汤(SZD)对KM小鼠的急性毒性及抗肿瘤活性, 为安全用药提供参考, 并对抗肿瘤效应进行初步评价。试验中, SPF级KM小鼠分别ig给予一定量的生理盐水(空白对照)和SZD, 测定其半数致死量(LD₅₀), 评价其对小鼠的毒性。另选取SPF级SD大鼠, ig给予SZD, 制备SZD含药学清(MS); MTT法、乳酸脱氢酶(LDH)活力检测, 甲基纤维素克隆形成实验, 流式细胞术等检测SZD-MS对肿瘤细胞增殖抑制及诱导凋亡情况。毒性测试结果显示, 小鼠无死亡, 无法得出SZD的LD₅₀。与空白对照组比较, 血液的生化学指标显示, 差异有统计学意义(P < 0.05, P < 0.01); H&E检测结果显示, SZD对小鼠的肝、肾功能有一定的损伤。抗肿瘤作用结果显示: 与溶剂对照组比较, SZD-MS对5种肿瘤细胞的增殖均有一定的抑制作用(P < 0.05, P < 0.01); SZD-MS对H22细胞LDH释放增加(P < 0.05); 细胞克隆的形成力度下降(P < 0.05, P < 0.01); 细胞的凋亡率增加(P < 0.01)。研究表明, SZD经口最大给药量为每只小鼠0.8 mL/d, 约相当于成人日剂量的120倍, 毒性较小, 安全性良好; 且有较好的抗腹水瘤活性。

关键词: 十枣汤, 急性毒性, 增殖抑制, 抗肿瘤活性

Abstract:

This study aimed to assess the acute toxicity of Shizao decoction (SZD) in KM mice and its antitumor activity, offering insights into drug safety and antitumor efficacy. In experiments, specific pathogen-free (SPF) KM mice were administered either saline (as a blank control) or SZD, and the half-lethal dose (LD₅₀) was determined. Additionally, SPF-grade SD rats were treated with SZD to produce SZD-medicated serum (SZD-MS). Assays, including the MTT method, lactic dehydrogenase (LDH) release, colony formation, and flow cytometry, were utilized to measure the inhibitory effects on cancer cell proliferation and induction of apoptosis. The toxicity tests revealed that none of the mice died after oral administration of SZD, rendering it impossible to establish an LD₅₀ value. Notably, serum biochemistry results significantly diverged from those of the blank control group (P < 0.05). Histopathology, using hematoxylin and eosin (H&E) staining, unveiled that SZD exerted detectable damage on the liver and kidneys of the mice. In terms of antitumor activity, SZD-MS demonstrated a significant inhibition of proliferation in five tumor cell lines when compared to the vehicle control (P < 0.05, P < 0.01). This finding was further supported by the increased LDH release from H22 cells (P < 0.05), a reduction in colony formation (P < 0.05, P < 0.01), and an elevated apoptosis rate (P < 0.01). In conclusion, the study revealed that the maximum oral dosage of SZD, set at 0.8 mL/d for each mouse (roughly 120 times the standard adult daily dose), presented minimal toxicity. Moreover, it possessed promising anti-ascite tumor activity, suggesting its safety and therapeutic potential.

Key words: Shizao decoction, Acute toxicity, Proliferation inhibition, Antitumor activity

Supporting:

马立威, 陈哲, 施国善, 王玉静, 陈颂, 倪世宇, 李京, 葛鹏玲, 刘吉成. 十枣汤急性毒性及抗肿瘤疗效评价[J]. 中国药学(英文版), 2024, 33(4): 329-338.

Liwei Ma, Zhe Chen, Guoshan Shi, Yujing Wang, Song Chen, Shiyu Ni, Jing Li, Pengling Ge, Jicheng Liu. Assessment of acute toxicity and antitumor efficacy of Shizao decoction[J]. Journal of Chinese Pharmaceutical Sciences, 2024, 33(4): 329-338.

图/表 10

Figure 1. Toxic effects of SZD on KM mice by intragastric administration. (A) The state of mice in the blank control and SZD groups was compared by intragastric administration; (B) The state of mice in the SZD group by intragastric administration; (C) Some mice emerged slight convulsion in the 0.4-mL SZD group.

Table 1. Effects of SZD on body weight in mice (x ± s, n = 8).

Table 2. Effects of SZD on organ coefficients in mice ((x ± s, n = 8).

Table 3. Effects of SZD on ALT, AST, Cr, and BUN contents in mice serum ((x ± s, n = 8).

Figure 2. Effects of SZD on histomorphology in liver and kidney of mice (×100). (A) Hepatic histomorphology of the blank control and 0.1-mL SZD groups; (B) Hepatic histomorphology of the 0.2-mL SZD group; (C) Hepatic histomorphology of the 0.4-mL SZD group; (D) Nephritic histomorphology of the blank control, 0.1-mL, and 0.2-mL SZD groups; (E) Nephritic histomorphology of the 0.4-mL SZD group.

Table 4. Effects of SZD-MS on cellular proliferation in 4T1, A549, HepG2, H22 and MCF-7 cells ((x ± s, n = 6).

Table 5. Effects of SZD-MS on LDH content in H22 cell ((x ± s, n = 3).

Table 6. Effects of SZD-MS on colony-forming efficiency in H22 cells ((x ± s, n = 3).

Figure 3. Effects of SZD-MS on apoptosis in H22 cells.

Table 7. Effects of SZD-MS on apoptosis rate in H22 cells ((x ± s, n = 3).

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十枣汤急性毒性及抗肿瘤疗效评价

Assessment of acute toxicity and antitumor efficacy of Shizao decoction

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