Please wait a minute...
Traditional Medicine Research  2017, Vol. 2 Issue (1): 27-32    DOI: 10.12032/TMR201705034
Basic Research     
Research on Xijiao Dihuang Decoction suppressing platelet apoptosis in immune-mediated aplastic anemia based on mitochondrial mediated pathway
Xia Le-Min, Cui Le-Le, Jiang Yi-Ling, Zheng Qin, Zhang Ai-Ping, Luo Mei-Hong*()
Department of Hematology, Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine, Baoshan Branch of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201999, China.
Download: HTML     PDF(285KB)
Export: BibTeX | EndNote (RIS)      

Highlights:

Excessive platelet apoptosis is one of the pathogenic cause for immune-induced aplastic anemia. In the present study, we established an immune-induced aplastic anemia model and gave Xijiao Dihuang Decoction lavage daily. The results of this study indicated Xijiao Dihuang Decoction could increase platelet number and prevent its apoptosis through the mitochondrial pathway.

Abstract

The aplastic anemia mice model was established in this study. BALB/c mice shall be treated with whole body irradiation with 60 Co-γradiation (5.5Gy, 1.1Gy/min × 5 min). then within 4 h, DBA/2 mice were injected lymphocyte suspension 1 × 106 cells /mouse through caudal vein. Grouping of testing animals: Normal control mice were healthy C57BL/6 mice without AA modeling. AA control group mice were exposed to radiation and cell transfusion and had no treatment with either CSA or XDD. CSA group mice received daily lavage with 0.027g/kg (0.1ml/10g) of CSA whereas XDD group mice received daily lavage with 19.5g/kg of XDD. The experimental result indicated CSA and XDD lavage mice had significantly higher platelet count and ΔΨm than AA mice (P < 0.05). Levels of Cyt C, PS and Ca2+ were significantly lower in CSA and XDD groups compare with AA group. More specifically, compared to CSA group, XDD group also had lower level of ΔΨm and higher level of Cyt C and Ca2+ (both P < 0.05). Both CSA and XDD treatment reduced Bak and Bax levels significantly compared to AA mice. However, XDD treatment still showed higher expressions than CSA (P < 0.05). CSA and XDD treatment increased the levels of Pro-apoptotic protein expressions ( caspase-8, caspase-3, caspase-9). XDD showed less potent effect than CSA in increasing these protein expressions (all P < 0.05). Therefore, we hypothesized XDD was possible to increase platelet number and prevent its apoptosis in immune-induced AA via the mitochondrial pathway.



Key wordsAplastic anemia      Xijiao Dihuang Decoction      Mitochondrial pathway      Apoptosis     
Published: 05 January 2017
Corresponding Authors: Luo Mei-Hong     E-mail: lmh021009@163.com
About author: §These authors contributed equally to this work. Executive Editor: Cui-Hong ZhuEnglish Editor: Xiao-Hui Liu
Cite this article:

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. Traditional Medicine Research, 2017, 2(1): 27-32. doi: 10.12032/TMR201705034

URL:

https://www.tmrjournals.com/tmr/EN/10.12032/TMR201705034     OR     https://www.tmrjournals.com/tmr/EN/Y2017/V2/I1/27

Group PLT(×109 /L)
Normal control 614.20±53.90
AA control 352.40±53.56 a
CSA group 580.30±88.86 b
XDD group 547.50±70.84 b
Table 1 Platelet count number in four groups of mice (\(\overline{x}\) ± SD)
Group ΔΨm % Cyt C % PS % Ca2+ %
Normal control 0.84±0.07 0.19±0.01 0.04±0.01 65.13±3.79
AA control 0.20±0.02 a 0.89±0.04 a 0.73±0.11a 340.02±25.64 a
CSA group 0.60±0.07 ab 0.33±0.05 ab 0.55±0.08 ab 96.49±5.95 ab
XDD group 0.33±0.03 abc 0.51±0.08 abc 0.49±0.20 ab 252.45±6.25 abc
Table 2 The mitochondrial transmembrane potential and biochemics in four groups of mice (\(\overline{x}\)±SD)
Group Bak Area/b-actin Bax Area/b-actin Caspase-3 Area/b-actin Caspase-8 Area/b-actin Caspase-9 Area/b-actin
Normal control 0.21±0.02 0.14±0.01 0.63±0.03 0.63±0.03 0.64±0.03
AA control 0.49±0.04a 0.52±0.04a 0.24±0.03a 0.24±0.02a 0.23±0.03a
CSA group 0.23±0.02ab 0.20±0.03ab 0.48±0.02ab 0.47±0.04ab 0.47±0.04ab
XDD group 0.35±0.04abc 0.40±0.03abc 0.39±0.06abc 0.40±0.07abc 0.40±0.04abc
Table 3 Pro-apoptotic protein expressions in four groups of mice (\(\overline{x}\)±SD)
Figure 1 Pro-apoptotic protein expressions by western blotting.
Note: A: normal control; B: AA control; C: CSA group; D: XDD group.
Figure 2 XDD inhibits platelet apoptosis in immune-induced bone marrow failure through mitochondrial pathway.
1.   Nakao S.Diagnostic problems in acquired aplastic anemia syndromes. Int J Hematol 2016, 104(2): 151-152.
2.   Valery L.Apoptosis in the anucleateplatelet. Blood Rev 2012, 26(2): 51-63.
3.   Leytin V, Allen DJ, Mutlu A, et al.Mitochondrial control of platelet apoptosis: effect of cyclosporin A, an inhibitor of the mitochondrial permeability transition pore. Lab Invest 2009, 89(4): 374-384.
4.   Wang Z, Cai F, Chen X, et al.The role of mitochondria-derived reactive oxygen species in hyperthermia-induced platelet apoptosis. Plos One 2013, 8(9): e75044.
5.   Liu HT, Zhao JM, Chu JX.Experimental Study of Low Dose Irradiation for Treatment of Immuno-Mediated Aplastic Anemia in Mice. J Exp Hematol 2007,15(3): 510-514.
6.   Yamazaki H.Acquired aplastic anemia. Rinsho Ketsueki 2016, 57(2): 91-97.
7.   Wang B, Zheng J.Platelet generation in vivo and in vitro. Springerplus 2016, 5(1): 787.
8.   Winkler J, Rand ML, Schmugge M, et al.Omi/HtrA2 and XIAP are components of platelet apoptosis signalling. ThrombHaemost 2013, 109(3): 532-539.
9.   Chen J, Desierto MJ, Feng X, et al.Immune-mediated bone marrow failure in C57BL/6 mice. Exp Hematol 2015, 43(4): 256-267.
10.   Satyamitra M, Ney P, Graves J III, et al.Mechanism of radioprotection by δ-tocotrienol: pharmacokinetics, pharmacodynamics and modulation of signalling pathways. Br J Radiol 2012, 85(1019): 93-103.
11.   Jagadish S, Rajeev N, NaveenKumar SK, et al. Platelet protective efficacy of 3, 4, 5 trisubstitutedisoxazole analogue by inhibiting ROS-mediated apoptosis and platelet aggregation. Mol Cell Biochem 2016, 414(1-2): 137-151.
[1] 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[J]. Traditional Medicine Research, 2019, 4(2): 99-108.
[2] 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]. Traditional Medicine Research, 2018, 3(3): 140-147.
[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]. Traditional Medicine Research, 2017, 2(3): 144-148.