Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (4): 414-422.doi: 10.3736/jcim20110411

• Original Experimental Research • Previous Articles     Next Articles

Inhibitory effects of traditional Chinese herbal medicine Tanreqing injection on proliferation of leukemia cells in vitro and the potential mechanisms

Bo Yang1, Xue-chun Lu1, Feng Zhang2, Yang Liu1, Hong-li Zhu1()   

  1. 1. Department of Geriatric Hematology, Chinese PLA General Hospital, Beijing 100853, China
    2. Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100029, China
  • Received:2010-09-28 Accepted:2010-12-28 Online:2011-04-20 Published:2011-04-15
  • Contact: Zhu Hong-li

Objective: The present study investigates the effects of Tanreqing injection, a compound Chinese herbal medicine, on the proliferation of leukemia cells in vitro and discusses the potential mechanisms.
Methods: Tanreqing injection was diluted to a series of concentrations (1∶2, 1∶4, 1∶8, 1∶16, 1∶32, 1∶64, 1∶128, 1∶256 and 1∶512) by volume and then independently applied to treat chronic myeloid leukemia K562 cells and T cell acute lymphocytic leukemia Molt4 cells at the proliferative stage. Cell growth was observed at different time intervals under a microscope. Cell proliferation was determined by cell counting kit-8 assay and the survival curve was delineated. The inhibitory rate and the half inhibitory concentration (IC50) were calculated. Molt4 cells were stained with propidium iodide (PI) and PI/Annexin V and then the cell cycle and apoptosis were analyzed by using flow cytometry. In addition, a real-time quantitative polymerase chain reaction was subjected to detect the expressions of apoptosis-related genes (bcl-2 and caspase-3) after Tanreqing treatment.
Results: Tanreqing injection had inhibitory effects on the proliferation of K562 cells and Molt4 cells. The most toxic concentrations were observed between 1∶2 and 1∶16 where cells were almost necrotic. The inhibitory effect manifested in a concentration- and time-dependent manner. The IC50 of K562 and Molt4 was 1∶333 and 1∶142, respectively. After 1∶32 Tanreqing injection treatment for 72 h, the number of Molt4 cells in the S phase significantly decreased (P<0.05), and the apoptosis rate markedly increased (P<0.05). In addition, increased caspase-3 expression and decreased bcl-2 expression were also observed (P<0.05).
Conclusion: Tanreqing injection can both inhibit the proliferation and promote the apoptosis of leukemia cells in vitro, whereby the potential mechanism seems to be mediated in part by decreasing S phase ratio, down-regulating bcl-2 expression and up-regulating caspase-3 expression.

Key words: Tanreqing injection, leukemia, K562 cells, Molt4 cells, cell proliferation, apoptosis

Figure 1

Growth curve of K562 cells after treatment with Tanreqing injection of different concentrations"


Figure 2

Growth curve of Molt4 cells after treatment with Tanreqing injection of different concentrations"


Figure 3

Microscopic observation of K562 cell growth after treatment with Tanreqing injection of different concentrations for 24 h (×100) At the concentrations of 1∶2 (B), 1∶4 (C), 1∶8 (D), and 1∶16 (E), cell death was obvious. At the concentrations of 1∶32 (F) and 1∶64 (G), cell growth was unfavorable. At the concentrations of 1∶128 (H), 1∶256 (I) and 1∶512 (J), cell growth was not significantly different from that in the control group (A)."

Figure 4

Microscopic observation of Molt4 cell growth after treatment with Tanreqing injection of different concentrations for 48 h (×100) At the concentrations of 1∶2 (B), 1∶4 (C), 1∶8 (D), and 1∶16 (E), cell death was obvious. At the concentrations of 1∶32 (F) and 1∶64 (G), cell growth was unfavorable. At the concentrations of 1∶128 (H), 1∶256 (I) and 1∶512 (J), cell growth was not significantly different from that in the control group (A)."

Figure 5

Cell cycle distribution of Molt4 cells treated with Tanreqing injection of different concentrations for 72h A: Control; B: 1∶512; C: 1∶256; D: 1∶128; E: 1∶64; F: 1∶32. “A” in the figures D, E and F represents apoptotic cells. The results show that cells in S phase gradually decreased and apoptotic cells appeared along with gradually increased concentration of Tanreqing injection."

Figure 6

Proportion of Molt4 cells in the S phase after treatment with Tanreqing injection of different concentrations for 72 h *P<0.05, vs control group."

Figure 7

Apoptotic Molt4 cells after treatment with Tanreqing injection of different concentrations for 72 h detected by flow cytometry after PI/Annexin V staining A: Control; B: 1∶512; C: 1∶256; D: 1∶128; E: 1∶64; F: 1∶32. The upper right quadrant represents cells positive for both Annexin V and PI; the lower right quadrant represents cells positive for Annexin V but negative for PI. PI-A: propidium iodide/Annexin V; FITC-A: fluorescein isothiocyanate-Annexin V."




[1] Wang CQ . Components and effects of Tanreqing injection[J]. Xian Dai Yi Yao Wei Sheng, 2006,22(5):720
王春全 . 痰热清注射液的方解及功效[J]. 现代医药卫生, 2006,22(5):720
[2] Zang YZ . Efficacy of Tanreqing injection in the treatment of fever in patients with myelodysplastic syndrome[J]. Zhong Yuan Yi Kan, 2006,33(7):79
doi: 10.3760/cma.j.issn.1674-4756.2006.07.056
臧玉柱 . 痰热清治疗骨髓增生异常综合征患者发热临床疗效观察[J]. 中原医刊, 2006,33(7):79
doi: 10.3760/cma.j.issn.1674-4756.2006.07.056
[3] Zhou Y, Wei YJ, Liu JF . Efficacy of Tanreqing injection in the treatment of pulmonary infection in patients with hematological malignancies[J]. Ji Ceng Yi Xue Lun Tan, 2006,10(20):882-883
周燕, 魏玉静, 刘建芳 . 痰热清治疗恶性血液病并肺部感染的疗效观察[J]. 基层医学论坛, 2006,10(20):882-883
[4] Zheng JS, Gu LG, Li PT, Zhao H . Effect of Tanreqing Injection on immune function of influenza virus FM1 infected mice[J]. Beijing Zhong Yi Yao Da Xue Xue Bao, 2006,29(11):756-759
doi: 10.3321/j.issn:1006-2157.2006.11.011
郑金粟, 顾立刚, 李澎涛, 赵慧 . 痰热清注射液对流感病毒FM1感染小鼠免疫功能影响的研究[J]. 北京中医药大学学报, 2006,29(11):756-759
doi: 10.3321/j.issn:1006-2157.2006.11.011
[5] Huang ZN, Huang ZY . Observation of Tanreqing injection in the prevention of concomitant respiratory tract infection in patients with hematological malignancies[J]. Youjiang Yi Xue, 2009,37(1):36-37
doi: 10.3969/j.issn.1003-1383.2009.01.018
黄枝宁, 黄枝优 . 痰热清注射液预防血液肿瘤患者并发呼吸道感染效果观察[J]. 右江医学, 2009,37(1):36-37
doi: 10.3969/j.issn.1003-1383.2009.01.018
[6] Ma JQ . Role of Tanreqing injection in the late stage radiotherapy of thoracic tumor[J]. Zhejiang Zhong Yi Za Zhi, 2006,41(6):369
doi: 10.3969/j.issn.0411-8421.2006.06.040
马建青 . 痰热清在胸部肿瘤后程放疗中的应用[J]. 浙江中医杂志, 2006,41(6):369
doi: 10.3969/j.issn.0411-8421.2006.06.040
[7] Zhao HL, Wang PY, Li W, Fan YJ, Wang B . Tanreqing injection in combination with antibiotics in the treatment of pulmonary infection in patients with tumor[J]. Yi Yao Lun Tan Za Zhi, 2005,26(21):60
doi: 10.3969/j.issn.1672-3422.2005.21.036
赵宏亮, 王鹏远, 李伟, 范英杰, 王彬 . 痰热清联合抗生素治疗肿瘤患者肺部感染[J]. 医药论坛杂志, 2005,26(21):60
doi: 10.3969/j.issn.1672-3422.2005.21.036
[8] Shi GJ, Sun HX . Treatment for 42 cases of carcinous fever by Tanreqing injection[J]. Henan Zhong Yi, 2005,25(9):25
史国军, 孙宏新 . 痰热清注射液治疗癌性发热42例[J]. 河南中医, 2005,25(9):25
[9] Meng M, Chen DZ, Wu BY, Xu WJ, Wu SH, Liu CY . Study of the effect of Tan Re Qing Injection on immune function of the mice infected with influenza virus FM1[J]. Hebei Da Xue Xue Bao Zi Ran Ke Xue Ban, 2006,26(5):529-535
doi: 10.3969/j.issn.1000-1565.2006.05.019
孟明, 陈冬志, 武变瑛, 许文杰, 吴素焕, 刘春颖 . 痰热清注射液对流感病毒感染小鼠免疫功能的影响[J]. 河北大学学报(自然科学版), 2006,26(5):529-535
doi: 10.3969/j.issn.1000-1565.2006.05.019
[10] Kuang JR, Yan P, Luo ZG . Application of Tanreqing in malignant tumor chemotherapy[J]. Yi Yao Lun Tan Za Zhi, 2007,28(5):42-43
况建荣, 燕平, 骆志国 . 痰热清在恶性肿瘤化疗中的应用[J]. 医药论坛杂志, 2007,28(5):42-43
[11] Zhang L . Efficacy of Tanreqing injection in combination with reduced glutathione in the treatment of liver dysfunction[J]. Zhongguo Zhong Yi Ji Zheng, 2008,17(10):1370-1372
张丽 . 痰热清联合还原型谷胱甘肽治疗肝功能损害疗效观察[J]. 中国中医急症, 2008,17(10):1370-1372
[12] Li PT, Zhang N, Zhu XL, Xu Y, Xie LJ, Wang B, Pang CH, Jia X, Wang XD . Protective effects of Tanreqing on lipopolysaccharide-induced acute lung injury in rats[J]. Zhongguo Yao Xue Za Zhi, 2005,40(7):518-521
doi: 10.3321/j.issn:1001-2494.2005.07.013
李澎涛, 张娜, 朱晓磊, 徐雅, 谢利军, 王冰, 庞春红, 贾旭, 王旭丹 . 痰热清注射液抗内毒素所致急性肺损伤的实验研究[J]. 中国药学杂志, 2005,40(7):518-521
doi: 10.3321/j.issn:1001-2494.2005.07.013
[13] Zhao XJ . Efficacy of Tanreqing injection in the treatment of cancerous fever[J]. Yi Yao Chan Ye Zi Xun, 2005,2(11):80
doi: 10.3969/j.issn.1673-7210.2005.11.071
赵新菊 . 痰热清注射液治疗癌性发热的疗效观察[J]. 医药产业资讯, 2005,2(11):80
doi: 10.3969/j.issn.1673-7210.2005.11.071
[14] Li W, Mao B, Wang G, Wang L, Chang J, Zhang Y, Wang MH, Guo J, Zheng YQ . Effect of Tanreqing Injection(痰热清注射液) on treatment of acute exacerbation of chronic obstructive pulmonary disease with Chinese medicine syndrome of retention of phlegm and heat in Fei(肺)[J]. Chin J Integr Med, 2010,16(2):131-137
doi: 10.1007/s11655-010-0131-y
[15] Tian YP, Hou MH, Li X . Effects of Tanreqing injection in the treatment of 52 cases of viral pneumonia[J]. Lin Chuang Fei Ke Za Zhi, 2004,9(5):505
田雅萍, 侯明惠, 李祥 . 痰热清注射液治疗病毒性肺炎52例临床观察[J]. 临床肺科杂志, 2004,9(5):505
[16] Costantini P, Jacotot E, Decaudin D, Kroemer G . Mitochondrion as a novel target of anticancer chemotherapy[J]. J Natl Cancer Inst, 2000,92(13):1042-1053
doi: 10.1093/jnci/92.13.1042 pmid: 10880547
[17] Reed JC . Double identity for proteins of the Bcl-2 family[J]. Nature, 1997,387(6635):773-776
doi: 10.1038/42867
[1] Cai-lian Fana, Wan-jun Caib, Meng-nan Ye, Miao Chen, Yi Dai. Qili Qiangxin, a compound herbal medicine formula, alleviates hypoxia-reoxygenation-induced apoptotic and autophagic cell death via suppression of ROS/AMPK/mTOR pathway in vitro. Journal of Integrative Medicine, 2022, 20(4): 365-375.
[2] Hong-xiao Li, Ling Shi, Shang-jie Liang Chen-chen Fang, Qian-qian Xu, Ge Lu, Qian Wang, Jie Cheng, Jie Shen, Mei-hong Shen. Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway. Journal of Integrative Medicine, 2022, 20(2): 163-172.
[3] Mohamed S. Othman, Sofian T. Obeidat, Amal H. Al-Bagawi, Mohamed A. Fareid, Alaa Fehaid, Ahmed E. Abdel Moneim. Green-synthetized selenium nanoparticles using berberine as a promising anticancer agent#br#
. Journal of Integrative Medicine, 2022, 20(1): 65-72.
[4] Hyongjun Jeon, Hee-Young Kim, Chang-Hwan Bae, Yukyung Lee, Sungtae Koo, Seungtae Kim. Korean red ginseng decreases 1-methyl-4-phenylpyridinium-induced mitophagy in SH-SY5Y cells. Journal of Integrative Medicine, 2021, 19(6): 537-544.
[5] Porwornwisit Tritripmongkol, Tullayakorn Plengsuriyakarn, Mayuri Tarasuk, Kesara Na-Bangchang. In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component, ethyl-p-methoxycinnamate, against cholangiocarcinoma. Journal of Integrative Medicine, 2020, 18(4): 326-333.
[6] Varuni Colamba Pathiranage, Ira Thabrew, Sameera R. Samarakoon, Kamani H. Tennekoon, Umapriyatharshini Rajagopalan, Meran K. Ediriweera. Evaluation of anticancer effects of a pharmaceutically viable extract of a traditional polyherbal mixture against non-small-cell lung cancer cells. Journal of Integrative Medicine, 2020, 18(3): 242-252.
[7] Maged Mohamed Maher Abou-Hashem, Dina Mohamed Abo-elmatty, Noha Mostafa Mesbah, Ahmed Mohamed Abd EL-Mawgoud. Induction of sub-G0 arrest and apoptosis by seed extract of Moringa peregrina (Forssk.) Fiori in cervical and prostate cancer cell lines. Journal of Integrative Medicine, 2019, 17(6): 410-422.
[8] Supakanya Kumkarnjana, Rutt Suttisri, Ubonthip Nimmannit, Apirada Sucontphunt, Mattaka Khongkow, Thongchai Koobkokkruad, Nontima Vardhanabhuti. Flavonoids kaempferide and 4,2′-dihydroxy-4′,5′,6′-trimethoxychalcone inhibit mitotic clonal expansion and induce apoptosis during the early phase of adipogenesis in 3T3-L1 cells. Journal of Integrative Medicine, 2019, 17(4): 288-295.
[9] Shriniwas S. Basaiyye, Sanjay Kashyap, Kannan Krishnamurthi, Saravanadevi Sivanesan. Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress (Lepidium sativum L.). Journal of Integrative Medicine, 2019, 17(3): 221-228.
[10] Carlos R. Oliveira, Daniel G. Spindola, Daniel M. Garcia, Adolfo Erustes, Alexandre Bechara, Caroline Palmeira-dos-Santos, Soraya S. Smaili, Gustavo J.S. Pereira, André Hinsberger, Ezequiel P. Viriato, Maria Cristina Marcucci, Alexandra C.H.F. Sawaya, Samantha L. Tomaz, Elaine G. Rodrigues, Claudia Bincoletto. Medicinal properties of Angelica archangelica root extract: Cytotoxicity in breast cancer cells and its protective effects against in vivo tumor development. Journal of Integrative Medicine, 2019, 17(2): 132-140.
[11] Syed Nasir Abbas Bukhari, Fahad Hussain, Hnin Ei Thu, Zahid Hussain. Synergistic effects of combined therapy of curcumin and Fructus Ligustri Lucidi for treatment of osteoporosis: cellular and molecular evidence of enhanced bone formation. Journal of Integrative Medicine, 2019, 17(1): 38-45.
[12] Bruno José Martins Da Silva, Sandro Wilson Gomes Pereira, Ana Paula Drummond Rodrigues, José Luiz Martins Do Nascimento, Edilene Oliveira Silva. In vitro antileishmanial effects of Physalis angulata root extract on Leishmania infantum. Journal of Integrative Medicine, 2018, 16(6): 404-410.
[13] Eric Wei Chiang Chan, Siu Kuin Wong, Hung Tuck Chan . Casticin from Vitex species: A short review on its anticancer and anti-inflammatory properties. Journal of Integrative Medicine, 2018, 16(3): 147-152.
[14] Jesmin Mondal, Asmita Samadder, Anisur Rahman Khuda-Bukhsh. Psorinum 6× triggers apoptosis signals in human lung cancer cells. Journal of Integrative Medicine, 2016, 14(2): 143-153.
[15] Qing-qing Zang, Lu Zhang, Ning Gao, Cheng Huang. Ophiopogonin D inhibits cell proliferation, causes cell cycle arrest at G2/M, and induces apoptosis in human breast carcinoma MCF-7 cells. Journal of Integrative Medicine, 2016, 14(1): 51-59.
Full text



[1] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[2] Hao Li, Ming-jiang Yao, Wen-ming Zhao, Jie Guan, Lin-lin Cai, Ling Cui. A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment. Journal of Chinese Integrative Medicine, 2008, 6(1): 25-31
[3] Zhi-chun Jin. Problems in establishing clinical guideline for integrated traditional Chinese and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 5-8
[4] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[7] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[8] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[9] Gui Yu, Jie Wang. Thinking on building the network cardiovasology of Chinese medicine. Journal of Chinese Integrative Medicine, 2012, 10(11): 1206-1210
[10] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239