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贵州中医药大学第一附属医院,贵阳 550001
安祯祥,博士,主任医师,从事中西医结合防治消化系统疾病的基础及临床研究,E-mail: 407206115@qq.com
何远利,主治医师,从事老年疾病研究,E-mail: 66587934@qq.com
收稿日期:2018-12-25,
网络出版日期:2019-05-07,
纸质出版日期:2019-12-05
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安祯祥, 何远利, 王敏, 等. 附子多糖对裸鼠胃癌移植瘤MMP-2,MMP-14表达的影响[J]. 中国实验方剂学杂志, 2019,25(23):79-85.
Zhen-xiang AN, Yuan-li HE, Min WANG, et al. Effect of Aconiti Lateralis Radix Praeparata Polysaccharide on Expressions of MMP-2, MMP-14 in Gastric Cancer Xenografts in Nude Mice[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(23): 79-85.
安祯祥, 何远利, 王敏, 等. 附子多糖对裸鼠胃癌移植瘤MMP-2,MMP-14表达的影响[J]. 中国实验方剂学杂志, 2019,25(23):79-85. DOI: 10.13422/j.cnki.syfjx.20191621.
Zhen-xiang AN, Yuan-li HE, Min WANG, et al. Effect of Aconiti Lateralis Radix Praeparata Polysaccharide on Expressions of MMP-2, MMP-14 in Gastric Cancer Xenografts in Nude Mice[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(23): 79-85. DOI: 10.13422/j.cnki.syfjx.20191621.
目的:
2
观察附子多糖对裸鼠胃癌移植瘤的生长抑制作用及对肿瘤组织基质金属蛋白酶-2(MMP-2),基质金属蛋白酶-14(MMP-14)表达的影响。
方法:
2
建立胃癌裸鼠移植瘤模型,随机分为模型组,5-氟尿嘧啶组(5-FU,0.01 g·kg
-1
),附子多糖高、低剂量组(0.2,0.1 g·kg
-1
),灌胃15 d。观察附子多糖对裸鼠胃癌瘤重的影响;光镜下观察肿瘤细胞的形态变化;酶联免疫吸附法(ELISA)测定血清中转化生长因子-
β
1
(TGF-
β
1
)含量;通过免疫组化、蛋白质免疫印迹法(Western blot)及实时荧光定量PCR(Real-time PCR)检测肿瘤组织中MMP-2,MMP-14蛋白及mRNA表达水平。
结果:
2
附子多糖高、低剂量组抑瘤率分别为52.83%,40.57%,与模型组比较,附子多糖高、低剂量组的瘤重显著降低(
P
<
0.01),附子多糖对裸鼠胃癌移植瘤的生长具有抑制作用;ELISA表明,与模型组比较,附子多糖高、低剂量组显著降低裸鼠胃癌血清TGF-
β
1
含量(
P
<
0.01);免疫组化表明,与模型组比较,附子多糖高、低剂量组MMP-2蛋白明显下调(
P
<
0.05,
P
<
0.01),MMP-14蛋白表达下降(
P
<
0.01);Western blot表明,与模型组比较,附子多糖高、低剂量组MMP-2,MMP-14蛋白表达下调(
P
<
0.01);Real-time PCR结果显示,与模型组比较,附子多糖高剂量组的MMP-2,MMP-14 mRNA表达下降(
P
<
0.05,
P
<
0.01)。
结论:
2
附子多糖可抑制裸鼠胃癌移植瘤的生长,抑制TGF-
β
1
表达,其抑瘤机制可能与下调MMP-2,MMP-14蛋白及mRNA的表达有关。
Objective:
2
To observe the growth inhibition effect of Aconiti Lateralis Radix Praeparata polysaccharide on the gastric cancer xenografts in nude mice and the expressions of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-14 (MMP-14) in tumor tissues.
Method:
2
The nude mice xenograft model was established and randomly divided into model group
5-fluorouracil group (5-FU
0.01 g·kg
-1
)
high and low-dose Aconiti Lateralis Radix Praeparata polysaccharide groups (0.2
0.1 g·kg
-1
). Each group was given drug by gavage for 15 days. The effect of Aconiti Lateralis Radix Praeparata polysaccharide on the weight of gastric cancer in nude mice was observed. Morphological changes of tumor cells were observed under light microscope. The content of transforming growth factor-
β
1
(TGF-
β
1
) in serum was determined by enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expressions of MMP-2
MMP-14 in tumor tissues were detected by immunohistochemistry
Western blot and Real-time PCR.
Result:
2
The tumor inhibition rates of high and low-dose Aconiti Lateralis Radix Praeparata polysaccharides were 52.83%
40.57%
respectively. Compared with model group
the tumor weight of high and low-dose Aconiti Lateralis Radix Praeparata polysaccharide was significantly lower (
P
<
0.01)
indicating the inhibitory effect on the growth of transplanted tumors. ELISA showed that compared with model group
Aconiti Lateralis Radix Praeparata polysaccharide could significantly reduce serum TGF-
β
1
content in nude mice (
P
<
0.01). Immunohistochemistry showed that compared with model group
MMP-2 protein was down-regulated in high and low-dose Aconiti Lateralis Radix Praeparata polysaccharide groups (
P
<
0.05
P
<
0.01)
and MMP-14 protein expression was decreased (
P
<
0.01). Western blot analysis showed that expressions of MMP-2 and MMP-14 were down-regulated in high and low-dose Aconiti Lateralis Radix Praeparata polysaccharide groups compared with model group (
P
<
0.01). Real-time PCR results showed that compared with model group
expressions of MMP-2 and MMP-14 mRNA in high-dose Aconiti Lateralis Radix Praeparata polysaccharide group decreased (
P
<
0.05
P
<
0.01).
Conclusion:
2
Aconiti Lateralis Radix Praeparata polysaccharide can inhibit the growth of gastric cancer xenografts in nude mice and the expression of TGF-
β
1
.The anti-tumor mechanism may be related to the down-regulation of MMP-2
MMP-14 protein and mRNA expressions.
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