Please wait a minute...
1Traditional Medicine Research  2019, Vol. 4 Issue (2): 99-108    DOI: 10.12032/TMR20190304108
Total saponins in Rubus parvifolius L. induce lymphoma cells apoptosis through upregulated Bax/Fas and downregulated Bcl-2 in vivo and in vitro
Xiao-Feng Xu1,*(), Ru-Bin Cheng2, Xue-Jin Zhang1, Rui-Lan Gao3
1 Department of Hematology, Zhejiang Provincial Integrated Chinese and Western medicine Hospital, Hangzhou, China.
2 College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou, China.
3 The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, China.
Download: HTML     PDF(749KB)
Export: BibTeX | EndNote (RIS)      


Total saponins of Rubus parvifolius L. (TSRP) induces Raji cell apoptosis by inhibiting Bcl-2, increasing Bax expression, and reducing Fas expression in vivo and in vitro. Additionally, TSRP has some effects on the chondriosome pathway of apoptosis in vitro.


Rubus parvifolius L. (RP) belongs to the family Rosaceae and is mainly produced in the temperate zone of the Northern hemisphere. As early as the Xihan Dynasty of China, Erya recorded that RP were edible. Bencao Shiyi recorded the medicinal properties of the root bark and fruit of RP, which include clearing heat, cooling blood, stopping bleeding, dispersing knots, relieving pain, diuresis, and detumescence.


Purpose: To investigate the effect of total saponins of Rubus parvifolius L. (TSRP) on lymphoma Raji cells and further discuss its mechanism. Methods: The model of nude mice bearing Raji cells was established, the volume, weight and inhibition rate of the transplanted tumor were analyzed and compared after different concentrations of TSRP treatment. Cell apoptosis and expression of Bcl-2, Bax, Fas proteins were detected by TUNEL and immunohistochemiscal method respectively. Effects of TSRP on cell proliferation were tested with MTT assay in vitro. Cell apoptosis and expression of Caspase-9, Caspase-3, Bcl-2, Bax and Fas proteins were tested with DAPI staining and Western blot. Results: TSRP significantly reduced the volume and tumor weight of Raji subcutaneous transplanted tumor and induced the apoptosis of Raji cells in vivo. The tumor inhibition rate of high-dose (100 mg/kg) TSRP is 90.84%. The TUNEL test results show that the fluorescence intensity of the tumor issue treated with TSRP is significantly improved. Compared with the control group, the fluorescence intensity of high-concentration TSRP is 82.43 ± 7.81, which is significantly different (P < 0.001). The results of immunohistochemistry test showed that the Bcl-2 expression of Raji cell treated with TSRP is obviously reduced, and Bax expression is obviously increased. Meanwhile, compared with that of control group, Fas expression is obviously reduced. MTT assay showed that TSRP can significantly inhibit proliferation of Raji cells with dose dependence. The inhibition rate of 400 μg/mL TSRP is 53.46 ± 4.90% (P < 0.001). DAPI staining results showed that TSRP can significantly induce cell apoptosis. According to Western blot results, it is found that TSRP can significantly inhibit activity of Bcl-2 and increase Bax expression, and TSRP can also inhibit Fas expression. Meanwhile, expression of Caspase-9 and Caspase-3 is also increased. Conclusion: TSRP could inhibit the proliferation of lymphoma via induction of apoptosis in a time and dose-dependent manner. Apoptotic signaling induced by TSRP was characterized by up-regulating Bax, Fas and Caspase-8 protein expression, and down-regulating of Bcl-2 protein expression.

Key wordsTotal Saponins of Rubus parvifolius L      Bcl-2      Bax      Fas      Apoptosis      Lymphoma     
Received: 28 December 2018      Published: 05 March 2019
Fund:  The study were supported by grants from Zhejiang Provincial Administration of Traditional Chinese Medicine (No. 2011ZA081, 2013ZB095 and 2015ZA147), Hangzhou Medical Science and Technology Plan (No. 2012A048).
Corresponding Authors: Xu Xiao-Feng     E-mail:
Cite this article:

Xiao-Feng Xu, Ru-Bin Cheng, Xue-Jin Zhang, Rui-Lan Gao. Total saponins in Rubus parvifolius L. induce lymphoma cells apoptosis through upregulated Bax/Fas and downregulated Bcl-2 in vivo and in vitro. 1Traditional Medicine Research, 2019, 4(2): 99-108. doi: 10.12032/TMR20190304108

URL:     OR

Groups Tumor weight (g) Size (mm3) Tumor inhibition rate (%)
Control 1.34 ± 0.50 2807.70 ± 128.43 -
TSRP (20 mg/kg) 0.86 ± 0.01 2022.59 ± 82.39# 27.96
TSRP (100 mg/kg) 0.21 ± 0.03* 257.29 ± 43.65# 90.84#
Table 1 Comparison of the tumor weight, size and tumor inhibition rate of each group after intragastric administration with TSRP
Groups Intragastric administration time (day)
8 11 14 17 20
Control 307.46 ± 13.83 456.57 ± 15.98 1093.29 ± 125.19 2080.01 ± 112.03 2807.70 ± 128.43
TSRP (20mg/kg) 212.51 ± 18.31 430.76 ± 10.28 998.43 ± 90.76 1122.59 ± 132.39# 2022 ± 182.39#
TSRP (100mg/kg) 207.22 ± 4.84 214.34 ± 8.32 235.45 ± 7.98# 243.15 ± 23.65# 257.29 ± 43.65#
Table 2 Tumor size change of the nude mice after intragastric administration (mm3)
Figure 1 Comparing Raji cell apoptosis of the transplanted tumor of each group with TUNEL analysis
TSRP, Total saponins of Rubus parvifolius L.. #, compared with the control group, P < 0.001.
Figure 2 Expression of Bcl-2, Bax and Fas of the transplanted tumor of each group (×200)
TSRP, Total saponins of Rubus parvifolius L..
Groups Absorbance Inhibition rate (%)
Control 0.780 ± 0.052 -
TSRP (100μg/ml) 0.639 ± 0.024 18.15 ± 3.33
TSRP (200μg/ml) 0.531 ± 0.034 32.21 ± 4.89
TSRP (400μg/ml) 0.366 ± 0.031 53.46 ± 4.90#
Table 3 Effects of TSRP at different concentrations with MTT
Figure 3 Morphology comparison after TSRP induces Raji cell apoptosis with DAPI analysis (fluorescence microscope × 200).
TSRP, Total saponins of Rubus parvifolius L..
Figure 4 Effect of TSRP at different concentrations on activity of Raji cell apoptosis controlling proteins Bcl-2, Bax, Fas, Casepase 9 and Casepase 3
Compared with the control group: #, P < 0.001; *, P = 0.027. TSRP, Total saponins of Rubus parvifolius L..
1.   Zheng YL, Hu CL.The experimental study of focal cerebral ischemia treated by Rubus parvifolius. Res Tradit Chin Med 2002, 18: 37.
2.   Ge YQ, Xu XF, Yang B, et al. Saponins from Rubus parvifolius L. induce apoptosis in human chronic myeloid leukemia cells through AMPK activation and STAT3 inhibition. Asian Pac J Cancer Prev 2014, 15: 5455-5461.
doi: 10.7314/APJCP.2014.15.13.5455 pmid: 25041018
3.   Zheng ZW, Zhang LJ, Huang QL, et al. Antitumor effect of total saponins of Rubus parvifolius L. on three cutaneous tumors in vitro. Chin J Dermatovenereol Integr Tradit West Med 2007, 6: 67-69.
4.   Evans BD, Smith IE, Shorthouse AJ, et al. A comparison of the response of human lung carcinoma xenografts to vindesine and vincristine. Br J Caner 1982, 45: 466-468.
doi: 10.1038/bjc.1982.75 pmid: 6280742
5.   Zhao ZJ, Chen Y, Francisco NM, et al. The application of CAR-T cell therapy in hematological malignancies: advantages and challenges. Acta Pharm Sin 2018, 8: 539-551.
doi: 10.1016/j.apsb.2018.03.001
6.   Wang ZG, Wu ZQ, Liu Y, et al. New development in CAR-T cell therapy. J Hematol Oncol 2017, 10: 53
doi: 10.1186/s13045-017-0423-1 pmid: 28222796
7.   Gravelle P, Burroni B, Péricart S, et al. Mechanisms of PD-1/PD-L1 expression and prognostic relevance in non-Hodgkin lymphoma: a summary of immunohistochemical studies. Oncotarget 2017, 8: 44960-44975.
doi: 10.18632/oncotarget.16680 pmid: 5546533
8.   Ilcus C, Bagacean C, Tempescul A, et al. Immune checkpoint blockade: the role of PD-1-PD-L axis in lymphoid malignancies. Onco Targets Ther 2017, 10: 2349-2363.
doi: 10.2147/OTT.S133385 pmid: 5417656
9.   Pérez-Garijo A, Steller H.Spreading the word: non-autonomous effects of apoptosis during development, regeneration and disease. Development 2015, 142: 3253-3262.
doi: 10.1242/dev.127878 pmid: 26443630
10.   Hosen N, Park CY, Tatsumi N, et al. CD96 is a leukemic stem cell-specific marker in human acute myeloid leukemia. Proc Natl Acad Sci USA 2007, 104: 11008-11013.
doi: 10.1073/pnas.0704271104 pmid: 17576927
11.   Brown EJ, Frazier WA.Integrin-associated protein (CD47) and its ligands. Trends Cell Bio 2001, 11: 130-135.
doi: 10.1016/S0962-8924(00)01906-1 pmid: 11306274
12.   12 .Wang Y,Krivtsov AV,Sinha AU,et al. The Wnt/b-Catenin Pathway Is Required for the Development leukemia stem cells in AML. Science 2010, 327: 1650-1653.
doi: 10.1126/science.1186624
13.   Koch U, Wilson A, Cobas M, et al. Simultaneous loss of beta- and gamma-catenin does not perturb hematopoiesis or lymphopoiesis. Blood 2008, 111: 160-164.
doi: 10.1182/blood-2007-07-099754 pmid: 17855627
14.   Yilmaz OH, Valdez R, Theisen BK, et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature 2006, 441: 475-482.
doi: 10.1038/nature04703
15.   Kharas MG, Okabe R, Ganis JJ, et al. Constitutively active AKT depletes hematopoietic stem cells and induces leukemia in mice. Blood 2010, 115: 1406-1415.
doi: 10.1182/blood-2009-06-229443 pmid: 20008787
16.   Kang Q, Zou H, Yang X, et al. Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma. Oncol Lett 2018, 15: 2161-2168.
doi: 10.3892/ol.2017.7570
17.   Lindqvist LM, Heinlein M, Huang DCS, et al. Prosurvival Bcl-2 family members affect autophagy only indirectly, by inhibiting Bax and Bak. Proc Natl Acad Sci USA 2014, 111: 8512-8517.
doi: 10.4161/auto.29639 pmid: 24912196
18.   Wang ZX, Fang JF, Xiao JZ.Correlation of the expression of inflammatory factors with expression of apoptosis -related genes Baxand Bcl-2, in burned rats. Exp Ther Med 2019, 17: 1790-1796.
19.   Gustafsson AB, Gottlieb RA.Bcl2-family members and apoptosis, taken to heart. Am J Physiol Cell Physiol 2007, 292: 45-51.
doi: 10.1152/ajpcell.00229.2006 pmid: 16943242
20.   Okada Y, Shimizu T, Maeno E, et al. Volume-sensitive chloride channels involved in apoptotic volume decrease and cell death. J Mem brBiol 2006, 209: 21-29.
doi: 10.1007/s00232-005-0836-6 pmid: 16685598
21.   21.Shimizu T,Numata T,Okada Y. A role of reactive oxygens peciesin apoptotic activation of volume-sensitive Cl (-) channel .Proc Natl Acad Sci USA,2004,101(17):6770-6773.
doi: 10.1073/pnas.0401604101 pmid: 15096609
22.   Kutuk O, Basaga H.Bcl2-protein family: implications in vascular apoptosis and atherosclerosis. Apoptosis 2006, 11: 1661-1675.
doi: 10.1007/s10495-006-9402-7 pmid: 16951924
23.   Liang Y, Nylander KD, Yan C, et al. Role of caspase-3-dependent Bcl-2 cleavage in potentiation of apoptosis by Bcl-2. Mol Pharmacol 2002, 61: 142-149.
doi: 10.1124/mol.61.1.142 pmid: 11752215
24.   Montes M, Coiras M, Becerra S, et al. Functional Consequences for Apoptosis by Transcription Elongation Regulator 1 (TCERG1)-Mediated Bcl-x and Fas/CD95 Alternative Splicing. PLoS One 2015, 10: e0139812.
doi: 10.1371/journal.pone.0139812 pmid: 26462236
25.   Tauber SC, Harms K, Falkenburger B, et al. Modulation of Hippocampal Neuroplasticity by Fas/CD95 Regulatory Protein 2 (Faim2) in the Course of Bacterial Meningitis. J Neuropathol Exp Neurol 2014, 73: 2-13.
doi: 10.1097/NEN.0000000000000020 pmid: 24335530
[1] Zheng-Bo Tao, Li-Yan Xiong, Li-Hui Wang, Chuan Zhang. Polysaccharide extracts of Cirsium japonicum protect rat H9c2 myocardial cells from oxidative stress induced by hydrogen peroxide[J]. 1Traditional Medicine Research, 2018, 3(3): 140-147.
[2] Jun-Jie Zhang, Xian-Zhi He, Guo-Shun Peng, Zhen-Kun Wang, Bin Ke, Jian Qin. Effect of alternate-day-fasting combined with Lingguizhugan Decoction on blood lipid profiles of hyperlipidemic rats[J]. 1Traditional Medicine Research, 2018, 3(3): 157-165.
[3] Nie Hai-Yang, Chen Rui, Zhang Hong-Na, Pan Zhi. Effects of saponin from the seed of Litchi chinensis Sonn on TGF-β1, FN and SOCS-1 in renal tubular epithelial cells under high glucose[J]. 1Traditional Medicine Research, 2017, 2(3): 144-148.
[4] Xia Le-Min, Cui Le-Le, Jiang Yi-Ling, Zheng Qin, Zhang Ai-Ping, Luo Mei-Hong. Research on Xijiao Dihuang Decoction suppressing platelet apoptosis in immune-mediated aplastic anemia based on mitochondrial mediated pathway[J]. 1Traditional Medicine Research, 2017, 2(1): 27-32.