传统中药王氏保赤丸的临床前毒理研究

doi:10.5246/jcps.2022.01.004

中国药学(英文版) ›› 2022, Vol. 31 ›› Issue (1): 31-46.DOI: 10.5246/jcps.2022.01.004

传统中药王氏保赤丸的临床前毒理研究

高涛¹, 刘蕾²^,^*(), 吴燕³^,^*(), 杜权¹^,^*()

1. 北京大学药学院天然及仿生药物国家重点实验室, 北京 100191
2. 北京医院药学部国家老年医学中心, 中国医学科学院老年医学研究院, 北京 100730
3. 中国人民解放军空军特色医学中心药剂科, 北京 100142

收稿日期:2021-09-20 修回日期:2021-10-21 接受日期:2021-11-05 出版日期:2022-01-12 发布日期:2022-01-13
通讯作者: 刘蕾, 吴燕, 杜权
作者简介:
+ Tel.: +86-10-82805780, E-mail: quan.du@pku.edu.cn
+ Tel.: +86-10-85133630, E-mail: liuleibjyy@126.com
+ Tel.: +86-10-66928796, E-mail: wuyan2001@163.com
基金资助:
Beijing Municipal Science & Technology Commission (Grant No. Z161100001816008), the National Natural Science Foundation of China (Grant No. 31571403), and Beijing Natural Science Foundation (Grant No. 2171001).

Preclinical safety evaluation of WangShiBaoChiWan

Tao Gao¹, Lei Liu²^,^*(), Yan Wu³^,^*(), Quan Du¹^,^*()

1 State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
2 Department of Pharmacy, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing 100730, China
3 Department of Pharmacy, Air Force Medical Center of PLA, Beijing 100142, China

Received:2021-09-20 Revised:2021-10-21 Accepted:2021-11-05 Online:2022-01-12 Published:2022-01-13
Contact: Lei Liu, Yan Wu, Quan Du

摘要/Abstract

摘要：

王氏保赤丸是一个具有180多年应用历史的中药复方。本研究利用大鼠给药模型, 对王氏保赤丸的临床前安全性进行了研究。在28天的给药实验期间, 对SD大鼠进行每日1次的灌胃给药, 对高、中、低剂量组而言, 给药剂量分别为60 mg/kg/day、600 mg/kg/day和1500 mg /kg/day。给药结束后, 进行15天的恢复期观察。以枸橼酸莫沙必利为参照药物, 给药剂量为37.5 mg/kg/day, 与王氏保赤丸的高剂量组保持一致。各剂量组在临床观察、摄食量、体重、脏器系数、血液生化、组织病理学等方面均未见不良反应, 并且也未见肠黑变现象的发生。在血液学实验中, 发现王氏保赤丸高剂量组的某些大鼠的血常规指标出现异常变化, 但在恢复期后基本回归正常, 这种可逆性的影响可能源于受损的大鼠肠道屏障。同时, 我们在枸橼酸莫沙必利给药组中也观察到了这一现象。除常规的组织病理学检测外, 我们还利用高通量基因测序技术进行了研究, 也未发现对细胞通路明显的毒性作用。综上所述, 实验动物在28天的给药期内, 对王氏保赤丸具有良好的耐受性。

关键词: 毒理研究, 中药, 高通量测序

Abstract:

WangShiBaoChiWan (WSBCW) is a traditional Chinese medicine with a recorded administration history of more than 180 years. In the present study, the preclinical safety of WSBCW was evaluated the preclinical safety of WSBCW using a toxicity test, which consisted of an administration period of 28 d and a recovery period of 15 d. During the test, male and female SD rats were administered the medicine once a day by oral gavage, at a dose of 60 mg/kg/day, 600 mg/kg/day, or 1500 mg/kg/day. As a reference medicine, mosapride citrate was administered at a dose of 37.5 mg/kg/day, which was clinically equivalent to the high-dosage treatment of WSBCW. With all the dosage groups, statistically, no adverse effect was observed in terms of clinical observation, food intake, body weights, organ coefficient, blood biochemistry, and histopathology examination. No intestinal melanosis was observed in the rats. When the data were examined animal by animal, test substance-related adverse effects were found with the high-dosage rats in hematology assay. The deranged, however, reversible changes suggested a compromised intestinal barrier, which was also observed with in mosapride citrate-treated rats. In addition to the histopathology assay, molecular toxicology was explored using high-throughput gene sequencing. No evident toxicity was revealed. In summary, administration of WSBCW was well tolerated within a treatment of 28 d.

Key words: Toxicology, Traditional Chinese medicine, Deep sequencing

Supporting:

高涛, 刘蕾, 吴燕, 杜权. 传统中药王氏保赤丸的临床前毒理研究[J]. 中国药学(英文版), 2022, 31(1): 31-46.

Tao Gao, Lei Liu, Yan Wu, Quan Du. Preclinical safety evaluation of WangShiBaoChiWan[J]. Journal of Chinese Pharmaceutical Sciences, 2022, 31(1): 31-46.

图/表 14

Figure 1. Body weights of WSBCW-treated rats in a toxicity test of 28 d. Ctrl, rats of the control group that were treated with saline; Low, rats of the low-dosage group that were treated with WSBCW of 60 mg/kg; Medium, rats of the medium-dosage group that were treated with WSBCW of 600 mg/kg; High, rats of the high-dosage group that were treated with WSBCW of 1500 mg/kg. Data are presented as mean ± SD (Student’s t-test, n = 6).

Figure 2. Food intakes of WSBCW-treated rats in a toxicity test of 28 d. Ctrl, rats of the control group that were treated with saline; Low, rats of the low-dosage group that were treated with WSBCW of 60 mg/kg; Medium, rats of the medium-dosage group that were treated with WSBCW of 600 mg/kg; High, rats of the high-dosage group that were treated with WSBCW of 1500 mg/kg. Data are presented as mean ± SD (Student’s t-test, n = 6).

Figure 3. Relative organ weights of WSBCW-treated rats. Weights of the organs are presented relative to the body weight.

Figure 4. Histopathology of WSBCW-treated rats. Representative photomicrographs of H&E stained sections of the liver and the jejunum of male mice (200× original magnification).

Figure 5. Effects on the length of intestinal villi. Quantitative results of intestinal villi length of rats under indicated treatments. The data are presented as mean ± SD (student’s t-test, n = 3); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns no significant.

Figure 6. Images of the colons of WSBCW-treated rats.

Table 1. Summary of serum biochemistry data of WSBCW-treated rats at the end of the administration period. ALB, albumin (reference ranges: 38-ic); ALP, alkaline phosphatase (40–442); ALT, alanine aminotransferase (22–137); AMY, amylase (502–1113); T-BIL, total bilirubin (4–7); BUN, blood urea nitrogen (2.5–6.6); CA, calcium (2.4–2.7); PHOS, phosphorus (0.74–2.26); CRE, creatinine (ic-53); GLU, glucose (6.6–13.7); NA+, sodium (133–144); K+, potassium (3.6–5.3); TP, total protein (66–88); GLOB, globulin (12–35); HEM, hemolysin. Data are presented as mean ± SD.

Table 2. Summary of serum biochemistry data of WSBCW-treated rats at the end of the recovery period.

Table 3. Summary of hematology data of WSBCW-treated rats at the end of the administration period. WBC, white blood cell count (reference ranges: 2.10–19.50); LYM, lymphocyte (2.00–14.10); MON, monocyte (0.00–0.98); NEU, neutrophil (0.10–5.40); RBC, red blood cell count (5.30–10.00); HGB, hemoglobin (14.0–18.0); HCT, hematocrit value (35.00–52.00); MCV, mean corpuscular volume (50–62); MCH, mean corpuscular hemoglobin (16.0–23.0); MCHC, mean corpuscular hemoglobin concentration (31.0–40.0); RDW, red blood cell distribution width; RDWs, red blood cell distribution width SD; PTL, platelet count; MPV, mean platelet volume; PCT, procalcitonin (500–1000); PDW, platelet distribution width; PDWs, platelet distribution width SD. Data are presented as mean ± SD.

Table 4. Summary of hematology data of WSBCW-treated rats at the end of the recovery period.

Figure 7. Analysis of hematology data of WSBCW-treated rats. Hematology data that deviate from the normal range are highlighted.

Figure 8. Comparative study of mosapride- and WSBCW-treated rats. Ctrl, rats of the control group that were treated with saline; WSBCW, rats of the high-dosage group that were treated with WSBCW of 1500 mg/kg; Mosapride, rats of the reference group that were treated with mosapride citrate of 37.5 mg/kg. Data are presented as mean ± SD (Student’s t-test, n = 6). (A) Body weights of the rats; (B) Food intakes; (C) Organ coefficients; (D) Hematology analysis.

Figure 9. Volcano map of the differentially regulated genes. The horizontal axis, fold-change of gene expression compared with the control; vertical axis, statistically difference of gene expression. Red dots indicate the significantly up-regulated genes, and green dots indicate the significantly down-regulated genes.

Figure 10. Comparative molecular toxicology of mosapride- and WSBCW-treated rats.

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传统中药王氏保赤丸的临床前毒理研究

Preclinical safety evaluation of WangShiBaoChiWan

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