摘要
目的:探究山慈菇含药血清(Pseudobulbus Cremastrae seu Pleiones pharmaceutic serum,PCPS)对肝癌细胞HepG2凋亡和上皮间质转化的影响。方法:选取40只雌性SD大鼠,随机分为空白组和山慈菇低、中、高剂量组,每组10只,山慈菇低(5 mL·kg^(-1))、中(10 mL·kg^(-1))、高(20 mL·kg^(-1))剂量组大鼠灌胃给予1 kg·L-1的山慈菇浓煎剂,空白组大鼠给予等量生理盐水,每天1次,连续6 d,腹主动脉取血制备含药血清,并用不同浓度的含药血清干预正常培养的HepG2细胞。CCK8法检测HepG2细胞的活力;EdU染色检测细胞增殖情况;流式细胞仪检测细胞凋亡情况;Transwell小室实验检测细胞侵袭能力;光镜下观察细胞上皮和间质的形态转换;蛋白免疫印迹法检测细胞周期抑制因子p27、凋亡抑制因子Sur-vivin及上皮间质转化标记物上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)、波形蛋白(Vimentin)的表达。结果:与空白组比较,10 mL·kg^(-1)、20 mL·kg^(-1) PCPS处理后,细胞生长曲线水平、细胞增殖率、侵袭率及Survivin、N-cadherin和Vimentin的表达水平显著下调(P<0.05),细胞凋亡率及p27、E-cadherin表达显著上调(P<0.05)。结论:PCPS可明显抑制肝癌细胞HepG2的生长,诱导凋亡,抑制其上皮间质转化。
Objective:To study the effect of Pseudobulbus Cremastrae seu Pleiones pharmaceutic serum(PCPS)on apoptosis and epithelial-mesenchymal transition of HepG2 cells.Methods:40 female SD rats were randomly divided into blank groups and low,medium,and high dose Shancigu groups,with 10 rats in each group.The low,medium and high dose Shancigu groups were given 1 kg·L-1 of Shancigu decoction by gavage at doses of 5,10,and 20 mL·kg^(-1),once a day for 6 consecutive days.And blood from the abdominal aorta was collected to prepare drug-containing serum,which was used to intervene the normal cultivated HepG2 cells.The viability of HepG2 cells was detected with CCK8,cell proliferation was detected with EdU staining,flow cytometry was used to detect cell apoptosis,and cell invasion ability was detected by the Transwell chamber experiment.The morphological transition of cell epithelium and mesenchyme was observed under the light microscope.And Western blotting was used to detect the levels of cell cycle inhibitor p27,apoptosis inhibitor Survivin,and epithelial-mesenchymal transition markers E-cadherin,N-cadherin,and Vimentin Expression.Results:Compared with the blank group,the levels of the cell growth curve,cell proliferation rate,invasion rate,and the expression levels of Survivin,N-cadherin and Vimentin were significantly decreased after 10 mL·kg^(-1) and 20 mL·kg^(-1) PCPS treatment,while the apoptosis rate and the expressions of p27 and E-cadherin were significantly increased(P<0.05).Conclusion:PCPS could obviously inhibit the growth of HepG2,induce apoptosis,and inhibit the epithelial-mesenchymal transition.
作者
程清波
杨艳萍
王滢
程文涛
张晋芬
张小莉
CHENG Qingbo;YANG Yanping;WANG Ying;CHENG Wentao;ZHANG Jinfen;ZHANG Xiaoli(Yingcheng People's Hospital,Yingcheng Hubei China 432400;The Affiliated Hospital of Wuhan University of Science and Technology,Wuhan Hubei China 430000)
出处
《中医学报》
CAS
2021年第10期2202-2207,共6页
Acta Chinese Medicine
基金
湖北省卫生和计划生育委员会科研项目(WJ2015MB133)。
关键词
山慈菇
肝癌细胞
生长运动
细胞凋亡
上皮间质转化
大鼠
Shancigu(Pseudobulbus Cremastrae Seu Pleiones)
hepatoma carcinoma cell
apoptosis
epithelial-mesenchymal transition
rat
