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西北民族大学 医学部,兰州 730000
宋易航,在读博士,讲师,从事妇科肿瘤及DNA损伤机制研究,E-mail:lichenghao222021@163.com
赵晋,博士,教授,从事细胞衰老及DNA损伤机制研究,E-mail:gz768@163.com
收稿日期:2021-09-01,
网络出版日期:2021-12-11,
纸质出版日期:2022-02-20
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宋易航,郭忠,窦春江等.黄芪多糖注射液对X射线治疗U14宫颈癌模型小鼠增效减毒作用[J].中国实验方剂学杂志,2022,28(04):84-90.
SONG Yi-hang,GUO Zhong,DOU Chun-jiang,et al.Effect Enhancing and Toxicity-reducing Activities of Astragalus Polysaccharide Injection on U14 Cervical Cancer in Model Mice Receiving X-Ray Treatment[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(04):84-90.
宋易航,郭忠,窦春江等.黄芪多糖注射液对X射线治疗U14宫颈癌模型小鼠增效减毒作用[J].中国实验方剂学杂志,2022,28(04):84-90. DOI: 10.13422/j.cnki.syfjx.20220421.
SONG Yi-hang,GUO Zhong,DOU Chun-jiang,et al.Effect Enhancing and Toxicity-reducing Activities of Astragalus Polysaccharide Injection on U14 Cervical Cancer in Model Mice Receiving X-Ray Treatment[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(04):84-90. DOI: 10.13422/j.cnki.syfjx.20220421.
目的
2
研究黄芪多糖注射液(APS)对X射线治疗U14宫颈癌模型小鼠的增效和减毒作用。
方法
2
体外培养U14小鼠宫颈癌细胞,注射接种于昆明小鼠右前肢腋下构建宫颈癌皮下荷瘤模型。将荷瘤小鼠随机分为模型组,X射线干预组(IR,6 Gy),APS组(APS,10 mL·kg
-1
·d
-1
),X射线联合黄芪多糖注射液组(IR+APS)。观察各组小鼠状态、体质量、进食量,并测量瘤体积大小变化;通过流式细胞术检测瘤体细胞周期和凋亡;蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)检测瘤组织凋亡相关蛋白p53,B淋巴细胞瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax)和剪切后半胱氨酸天冬氨酸蛋白水解酶-3(cleaved Caspase-3)蛋白和mRNA水平的变化。
结果
2
与模型组比较,IR组小鼠精神状态不佳,活动度减少,第8天开始,IR组和IR+APS组进食量降低、体质量降低(
P
<
0.05,
P
<
0.01),第9天开始,IR组和IR+APS组小鼠瘤体积更小(
P
<
0.01),IR组小鼠瘤组织中细胞G
1
期比例升高,S期比例降低(
P
<
0.01),IR+APS组小鼠瘤组织中细胞G
1
比例升高,G
2
和S期比例降低(
P
<
0.05,
P
<
0.01),IR组和IR+APS组凋亡率显著增加(
P
<
0.01)。与模型组比较,IR组和IR+APS组瘤组织中cleaved Caspase-3和Bax蛋白和mRNA表达均升高,Bcl-2和p53蛋白和mRNA表达降低(
P
<
0.05,
P
<
0.01);与IR组比较,IR+APS组小鼠活动度和毛发较好,小鼠体质量和进食量升高(
P<
0.05),IR+APS组瘤体积减小(
P<
0.05),瘤组织中细胞G
2
期降低,S期升高(
P
<
0.05),凋亡率增加(
P
<
0.05),瘤组织中Bax蛋白表达均升高,Bcl-2和p53蛋白表达降低(
P
<
0.05,
P
<
0.01)。
结论
2
APS能够维持X射线治疗U14宫颈癌模型小鼠生命状态稳定,促进肿瘤细胞凋亡,具有增效和减毒的作用。
Objective
2
To study the effect-enhancing and toxicity-reducing activities of astragalus polysaccharide injection (APS) on U14 cervical cancer in model mice receiving X-ray treatment.
Method
2
U14 mouse cervical cancer cells were cultured
in vitro
and injected into the right forelimb armpit of Kunming mice for constructing a subcutaneous tumor-bearing model of cervical cancer. The tumor-bearing mice were randomly divided into the model group, X-ray intervention(IR, 6 Gy) group, APS (10 mL·kg
-1
·d
-1
) group, and IR + APS group. Following the observation of the state, body mass, and food intake of mice in each group, the volume of the tumor was measured. The tumor cell cycle and apoptosis were determined by flow cytometry. The protein and mRNA expression levels of apoptosis-related proteins p53, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and cleaved cysteine-dependent aspartate-directed protease-3 (Caspase-3) in tumor tissues were assayed by Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR).
Result
2
The comparison with the model group showed that mice in the IR group had poor mental status and reduced mobility. The IR group and IR + APS group exhibited reduced food intake and body mass since the 8
th
d (
P
<
0.05,
P
<
0.01) and narrowed tumor volume since the 9
th
d (
P
<
0.01). In the IR group, the proportion of cells in the G
1
phase was increased, while the proportion of those in the S phase was decreased (
P
<
0.01). In the IR + APS group, the proportion of cells in the G
1
phase rose, whereas the proportion of those in the G
2
and S phases cells declined (
P
<
0.05,
P
<
0.01). The apoptotic rates in both the IR group and IR + APS group were elevated significantly (
P
<
0.01). Compared with the model group, the IR group and IR + APS group displayed up-regulated cleaved Caspase-3 and Bax protein and mRNA expression in tumor tissues, but down-regulated Bcl-2 and p53 protein and mRNA expression (
P
<
0.05,
P
<
0.01). Compared with the IR group, the mice in the IR + APS group had better mobility and hair, normal body mass, and increased food intake (
P
<
0.05). The tumor volume in the IR + APS group was reduced (
P
<
0.05). The proportion of cells in the G
2
phase was reduced, but the proportion of those in the S phase was raised (
P
<
0.05). The apoptosis rate was increased (
P
<
0.05). The apoptosis-related protein Bax protein expression in the tumor tissue was up-regulated, while the protein expression levels of Bcl-2 and p53 were down-regulated (
P
<
0.05,
P
<
0.01).
Conclusion
2
APS maintains the life state of U14 cervical cancer model mice treated with X-ray and promotes tumor cell apoptosis, thus enhancing the efficiency and reducing toxicity.
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