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引用本文:银艳桃,王建超,廖娟,玉叶,朱朝阳,邓鑫.肝宁方含药血清对人肝星状细胞增殖及凋亡的影响[J].中国现代应用药学,2020,37(3):269-274.
YIN Yantao,WANG Jianchao,LIAO Juan,YU Ye,ZHU Chaoyang,DENG Xin.Effect of Ganning Formula on Proliferation and Apoptosis of Human Hepatic Stellate Cells[J].Chin J Mod Appl Pharm(中国现代应用药学),2020,37(3):269-274.
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肝宁方含药血清对人肝星状细胞增殖及凋亡的影响
银艳桃1, 王建超1, 廖娟1, 玉叶1, 朱朝阳2, 邓鑫1,2
1.广西中医药大学, 南宁 530000;2.广西中医药大学附属瑞康医院, 南宁 530011
摘要:
目的 研究肝宁方含药血清对人肝星状细胞(HSC-LX2)增殖及凋亡的影响。方法 40只SD大鼠随机分为4组,每组10只,空白对照组与TGF-β1组SD大鼠给予生理盐水灌胃,TGF-β1+肝宁方组及肝宁方组SD大鼠给予肝宁方灌胃给药(7 mL·kg-1·d-1),连续灌胃10 d后取血离心,制备干燥血清粉末备用。将HSC-LX2分为空白对照组、TGF-β1组、TGF-β1+肝宁方组及肝宁方组。根据分组加入相应的20%大鼠血清对HSC-LX2进行培养造模。CCK8检测各组HSC-LX2细胞增殖率;流式细胞术检测各组HSC-LX2细胞的周期和细胞凋亡;qRT-PCR以及Western blotting分别检测各组α-SMA、Samd2、Samd3、Samd4、Samd7 mRNA及蛋白的表达。结果 CCK8结果显示,TGF-β1+肝宁方组及肝宁方组HSC-LX2增殖受到抑制;流式细胞术结果显示,TGF-β1+肝宁方组及肝宁方组HSC-LX2周期停滞在G0/G1期并能促进细胞凋亡;qRT-PCR及Western blotting结果显示,TGF-β1+肝宁方组及肝宁方组α-SMA、Samd2、Samd3、Samd4 mRNA及蛋白的表达受到抑制,Samd7 mRNA及蛋白的表达增强。结论 肝宁方具有抑制HSC-LX2增殖、促进HSC-LX2凋亡的作用,其作用与调控TGF-β1/Samd信号通路相关。
关键词:  肝宁方  肝星状细胞  增殖  凋亡  TGF-β1/Samd
DOI:10.13748/j.cnki.issn1007-7693.2020.03.003
分类号:R285.5
基金项目:国家科技重大专项(2018ZX10303501);国家自然科学基金项目(81860790,81360532);广西医疗卫生适宜技术研究与开发项目(S201503);广西研究生教育创新课题(xjyb151)
Effect of Ganning Formula on Proliferation and Apoptosis of Human Hepatic Stellate Cells
YIN Yantao1, WANG Jianchao1, LIAO Juan1, YU Ye1, ZHU Chaoyang2, DENG Xin1,2
1.Guangxi University of Chinese Medicine, Nanning 530000, China;2.Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530011, China
Abstract:
OBJECTIVE To study the effect of Ganning formula on proliferation and apoptosis of human hepatic stellate cells(HSC-LX2). METHODS Forty SD rats were randomly divided into 4 groups, 10 in each group. The blank control group and the TGF-β1 group were given normal saline gavage, and the TGF-β1+Ganning formula group and Ganning formula group were given gavage(7 mL·kg-1·d-1). HSC-LX2 was divided into blank control group, TGF-β1 group, TGF-β1+Ganning formula group and Ganning formula group. The corresponding rat serum was added to the corresponding groups according to the data for the culture and modeling of HSC-LX2. The proliferation of HSC-LX2 cells in each group was detected by CCK8. Flow cytometry was used to detect the cell cycle and apoptosis of HSC-LX2 cells in each group. The mRNA and protein expressions of α-SMA, Samd2, Samd3, Samd4 and Samd7 were detected by qRT-PCR and Western blotting. RESULTS The results of CCK8 showed that the proliferation of HSC-LX2 was inhibited in the serum group containing Ganning formula. The results of flow cytometry showed that the cycle of HSC-LX2 in the G0/G1 phase of the serum group containing Ganning formula was stagnated and cell apoptosis was promoted. qRT-PCR and Western blotting results showed that the mRNA and protein expressions of α-SMA, Samd2, Samd3 and Samd4 were inhibited and the mRNA and protein expressions of Samd7 were enhanced. CONCLUSION Ganning formula can inhibit the proliferation of HSC-LX2 and promote the apoptosis of HSC-LX2, which may be related to the regulation of TGF-β1/Samd signaling pathway.
Key words:  Ganning formula  hepatic stellate cells  proliferation  apoptosis  TGF-β1/Samd
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