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南昌市第三医院,南昌 330009
白羽,硕士,副主任医师,从事临床血液学检验研究,E-mail:by444826970@126.com
收稿日期:2021-05-21,
网络出版日期:2022-05-30,
纸质出版日期:2023-02-05
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白羽,熊文祺,朱春龙等.虎杖苷对骨髓瘤细胞的生长、凋亡及ROS/p38 MAPK信号通路作用[J].中国实验方剂学杂志,2023,29(03):104-109.
BAI Yu,XIONG Wenqi,ZHU Chunlong,et al.Effect of Polydatin on Growth,Apoptosis, and ROS/p38 MAPK Signaling Pathway of Myeloma Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(03):104-109.
白羽,熊文祺,朱春龙等.虎杖苷对骨髓瘤细胞的生长、凋亡及ROS/p38 MAPK信号通路作用[J].中国实验方剂学杂志,2023,29(03):104-109. DOI: 10.13422/j.cnki.syfjx.20230393.
BAI Yu,XIONG Wenqi,ZHU Chunlong,et al.Effect of Polydatin on Growth,Apoptosis, and ROS/p38 MAPK Signaling Pathway of Myeloma Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(03):104-109. DOI: 10.13422/j.cnki.syfjx.20230393.
目的
2
研究虎杖苷对骨髓瘤细胞生长、凋亡及ROS/p38 MAPK信号通路的作用。
方法
2
体外培养人多发性骨髓瘤(MM)细胞系U266细胞,通过细胞增殖与活性检测(CCK-8)法检测0、20、40、80、160、200 mg·L
-1
质量浓度虎杖苷对U266细胞生长的影响,计算其半抑制浓度(IC
50
)。取处于对数生长期的U266细胞,随机分为空白组、虎杖苷低质量浓度(80 mg·L
-1
)组、虎杖苷高质量浓度(160 mg·L
-1
)组、硼替佐米(75 nmol·L
-1
)组,经药物分组处理后,通过CCK-8法测定各组细胞活力;通过流式细胞实验测定各组细胞凋亡率;通过酶联免疫吸附测定法(ELISA)测定各组细胞炎性因子肿瘤坏死因子-
α
(TNF-
α
)、白细胞介素-1
β
(IL-1
β
)及活性氧(ROS)水平;通过蛋白免疫印迹法检测各组细胞凋亡相关因子胱天蛋白酶-9(Caspase-9)、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)及p38丝裂原活化蛋白激酶(MAPK)、磷酸化(p)-p38 MAPK蛋白表达水平。
结果
2
与空白组比较,不同浓度虎杖苷均可抑制U266细胞生长(
P
<
0.05),并随浓度升高而作用增强,呈剂量依赖性,IC
50
为156.54 mg·L
-1
。与空白组比较,虎杖苷低、高质量浓度组和硼替佐米组细胞活力明显降低(
P
<
0.05),凋亡率、TNF-
α
、IL-1
β
及ROS水平、细胞Caspase-9、Bax蛋白表达水平及p-p38 MAPK/p38 MAPK水平明显增高(
P
<
0.05)。与虎杖苷低质量浓度组比较,虎杖苷高质量浓度组和硼替佐米组上述指标改善更明显(
P
<
0.05),虎杖苷高质量浓度组和硼替佐米差异无统计学意义。
结论
2
虎杖苷可激活ROS/p38 MAPK信号通路,促使炎性因子表达,抑制U266细胞生长,促进其凋亡。
Objective
2
To analyze the effects of polydatin on myeloma cell growth,apoptosis, and reactive oxygen species(ROS)/p38 mitogen-activated protein kinase(MAPK) signaling pathway.
Method
2
Human multiple myeloma (MM) cell line U266 cells were cultured in vitro,and the effects of polydatin at 0,20,40,80,160,200 mg·L
-1
on the growth of U266 cells were detected by cell counting kit-8(CCK-8)assay. The half-maximal inhibitory concentration(IC
50
)was calculated. U266 cells in the logarithmic growth phase were randomly divided into a control group, low- and high-dose polydatin (80 and 160 mg·L
-1
) groups, and a bortezomib (75 nmol·L
-1
) group. After treatment with corresponding drugs,the cell viability of each group was determined by CCK-8 assay. The apoptosis rate of each group was measured by flow cytometry. The levels of inflammatory factors, such as tumor necrosis factor-
α
(TNF-
α
),interleukin-1
β
(IL-1
β
), and ROS in each group were measured by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of apoptosis-related factors, including cysteine aspartate-specific protease-9(Caspase-9),B-cell lymphoma-2-associated X protein(Bax),p38 MAPK,and phosphorylated (p)-p38 MAPK in each group were detected by Western blot.
Result
2
Compared with the results in the control group, polydatin of different concentrations could inhibit the growth of U266 cells (
P
<
0.05),and the effect was potentiated with the increase in the concentration,with IC
50
of 156.54 mg·L
-1
. Compared with the control group,the groups with drug treatment showed blunted cell viability (
P
<
0.05) and increased apoptosis rate,TNF-
α
,IL-1
β
,and ROS levels, protein expression levels of Caspase-9, Bax,and p-p38 MAPK/p38 MAPK (
P
<
0.05). Compared with the low-dose polydatin group, the high-dose polydatin group and the bortezomib group showed improved indicators mentioned above (
P
<
0.05), and there was no significant difference between the high-dose polydatin group and the bortezomib group.
Conclusion
2
Polydatin can activate the ROS/p38 MAPK signaling pathway,promote the expression of inflammatory factors,inhibit the growth of U266 cells,and promote their apoptosis.
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