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消癥汤对大鼠子宫平滑肌瘤细胞增殖和迁移的影响及机制
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| 篇名： | 消癥汤对大鼠子宫平滑肌瘤细胞增殖和迁移的影响及机制 |
| TITLE： | Effects and Mechanism of Xiaozheng Decoction on the Proliferation and Migration of Uterine Leiomyoma Cells in Rat |
| 摘要： | 目的：初步研究消癥汤对大鼠子宫平滑肌瘤细胞增殖和迁移的影响及作用机制。方法：将雌性SD大鼠随机分为正常对照组、模型组、化学药阳性对照组（米非司酮片2.25mg/kg）、中药阳性对照组（桂枝茯苓胶囊200mg/kg）和消癥汤低、中、高剂量组（1.4、2.8、5.6g/kg），每组15只。除正常对照组外，其余各组大鼠均肌内注射雌激素和孕激素建立子宫平滑肌瘤模型。自建模第2天起，各给药组大鼠灌胃相应药液，正常对照组和模型组大鼠灌胃等体积生理盐水，每天1次，连续4个月。末次给药后，摘取子宫并观察其形态，计算子宫系数；分离培养子宫平滑肌瘤细胞或子宫平滑肌细胞，采用MTT法检测细胞增殖率，细胞划痕实验检测子宫平滑肌瘤细胞迁移率，流式细胞术检测细胞凋亡率，荧光定量聚合酶链反应技术检测细胞中高迁移率族蛋白B1（HMGB1）mRNA表达情况，酶联免疫吸附测定法检测细胞中磷酸化蛋白激酶B（p-Akt）、核因子κB抑制蛋白α（IκBα）、磷酸化转化生长因子β激活激酶1（p-TAK1）的表达水平，Westernblot法检测细胞质内HMGB1、磷脂酰肌醇3激酶（PI3K）、p-Akt蛋白和细胞核内核因子κBp65（NF-κBp65）蛋白的表达水平。结果：与正常对照组比较，模型组大鼠子宫肌层明显增厚，子宫平滑肌细胞明显增多且大小不一，部分区域的肌纤维排列紊乱，子宫系数和细胞中HMGB1mRNA及其蛋白的相对表达水平均显著升高（P＜0.01）。与模型组比较，消癥汤各剂量组以及各阳性对照组大鼠的子宫平滑肌层增厚等情况均有不同程度改善，子宫系数以及子宫平滑肌瘤细胞增殖率、相对迁移率和细胞中HMGB1mRNA及其蛋白的相对表达水平，p-Akt、IκBα的表达水平，细胞质中PI3K、p-Akt蛋白的相对表达水平（除消癥汤低剂量组外）均显著降低（P＜0.05或P＜0.01），子宫平滑肌瘤细胞凋亡率和细胞中p-TAK1的表达水平、细胞核内NF-κBp65蛋白的相对表达水平均显著升高（P＜0.05或P＜0.01），且消癥汤的作用有剂量依赖的趋势。结论：消癥汤可能通过下调HMGB1、PI3K、p-Akt表达，上调NF-κBp65表达，进而抑制子宫平滑肌瘤细胞的增殖和迁移，并促使其凋亡。 |
| ABSTRACT： | OBJECTIVE：To study the effects a nd mechanism of X iaozheng decoction on the proliferation and migration of uterine leiomyoma cells in rat. METHODS ：Female SD rats were randomly divided into normal control group ，model group ， chemical medicine positive control group （Mifepristone tablets ，2.25 mg/kg），TCM positive control group （Guizhi fuling capsules ， 200 mg/kg），Xiaozheng decoction low-dose ，medium-dose and high-dose groups （1.4，2.8，5.6 g/kg），with 15 rats in each group. Except for normal control group ，other groups were given intramuscular injection of estrogen and progesterone to induce uterine leiomyomas model. On the second day after modeling ，rats in administration groups were given relevant medicine intragastrically ； normal control group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 months. After last administration ，the uterus was removed and its morphology was observed ；the uterine coefficient was calculated. Uterine leiomyoma cells or uterine smooth muscle cells were isolated and cultured. The proliferation rate and migration rate of cells were detected by MTT method and cell scratch test ；flow cytometry was used to detect the cell apoptosis rate ；mRNA expression of HMGB 1 were detected by RT-qPCR. The expression of phosphorylated protein kinase B （p-Akt），nuclear factor κB inhibitor α （IκBα）and phosphorylated transforming growth factor β-activated kinase 1（p-TAK1）were detected by ELISA ；the protein expression of HMGB 1，phospholipid 3 kinase（PI3K），p-Akt in cytoplasm and nuclear factor κB p65（NF-κB p65）in nucleus were detected by Western blot assay. RESULTS ：Compared with normal control g roup，the myometrium of the model group was significantly thickened ，the number of uterine smooth muscle cells were significantly increased and the sizes were 83777930。E-mail：jianghua_ouyang@126.com different，the arrangement of muscle fibers in so me areas was disordered，the uterine coefficient ，and the relative expression of HMGB 1 mRNA and protein were increased significantly （P＜ 0.01）. Compared with model group ，the thickening of uterine myometrium and other symptoms were improved to different extents in Xiaozheng decoction groups and positive control groups ；the uterine coefficient ，cell proliferation rate ，migration rate ，mRNA and protein expression of HMGB 1，the expression of p-Akt and IκBα，protein expression of PI 3K and p-Akt （except for Xiaozheng decoction low-dose group ）in cytoplasm were all decreased significantly （P＜0.05 or P＜0.01）；the cell apoptosis rate ，the expression of p-TAK 1，protein expression of NF-κB p65 in nucleus were all increased significantly （P＜0.05 or P＜0.01）. The effects of Xiaozheng decoction showed a dose-dependent trend. CONCLUSIONS ：Xiaozheng decoction can inhibit the proliferation and migration of uterine leiomyoma cells by down-regulating the expression of HMGB 1，PI3K and p-Akt ，up-regulating the expression of NF-κB p65，so as to promote cell apoptosis. |
| 期刊： | 2021年第32卷第20期 |
| 作者： | 吴艳丽，范楷，孔守芳，赵秀芳，欧阳江华 |
| AUTHORS： | WU Yanli，FAN Kai，KONG Shoufang ，ZHAO Xiufang ，OUYANG Jianghua |
| 关键字： | 消癥汤；子宫平滑肌瘤；高迁移率族蛋白B1；磷脂酰肌醇3激酶/蛋白激酶B；核因子κB；大鼠 |
| KEYWORDS： | Xiaozheng decoction ；Uterine leiomyoma ；HMGB1；PI3K/Akt；NF-κB；Rat |
| 阅读数： | 202 次 |
| 本月下载数： | 3 次 |
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