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苦参碱对乙醛活化的肝星状细胞CFSC-8B增殖和胶原合成的影

响及机制研究 Δ

王晓丽（齐齐哈尔医学院药学院，黑龙江 齐齐哈尔 161006）
＊

中图分类号 R962 文献标志码 A 文章编号 1001-0408（2020）11-1353-06
DOI 10.6039/j.issn.1001-0408.2020.11.12

摘 要 目的：研究苦参碱对乙醛活化的大鼠肝星状细胞CFSC-8B增殖和胶原合成的影响，并探讨其可能的作用机制。方法：取
体外培养的 CFSC-8B 细胞，分为空白对照组、模型组、阳性对照组（2.5 μmol/L 秋水仙碱）和苦参碱低、中、高浓度组（30、60、120
μmol/L）。除空白对照组外，其余各组细胞均加入200 μmol/L乙醛溶液诱导活化，并同时加入相应药液（空白对照组和模型组加入
等体积空白培养液），共同作用 24 h 后，采用 CCK-8 法检测各组细胞的存活率。另取细胞分为空白对照组、模型组、阳性对照组
（2.5 μmol/L秋水仙碱）和苦参碱中、高浓度组（60、120 μmol/L），同法活化和加药处理。分别采用酶消化法检测细胞培养液中羟脯
氨酸（Hyp）含量；采用酶联免疫吸附法检测细胞培养液中Ⅰ型胶原蛋白（Col-Ⅰ）和Ⅲ型胶原蛋白（Col-Ⅲ）含量；采用实时荧光定
量-聚合酶链式反应法检测细胞中α-平滑肌激动蛋白（α-SMA）、转化生长因子β 1 （TGF-β 1 ）、TGF-β 1Ⅰ型受体（TβR-Ⅰ）、TGF-β 1Ⅱ型
受体（TβR-Ⅱ）、Smad3、Smad4 和 Smad7 mRNA 表达水平；采用 Western blotting 法检测细胞中α-SMA、TGF-β 1、TβR-Ⅰ、TβR-Ⅱ、
Smad3、Smad4 和 Smad7 蛋白表达水平。结果：与空白对照组比较，模型组细胞存活率显著升高（P＜0.05）；细胞培养液中 Hyp、
Col-Ⅰ、Col-Ⅲ含量和细胞中α-SMA、TGF-β 1、TβR-Ⅰ、TβR-Ⅱ、Smad3和Smad4 mRNA及其蛋白表达水平均显著升高（P＜0.05），
细胞中Smad7 mRNA及其蛋白表达水平均显著降低（P＜0.05）。与模型组比较，阳性对照组和苦参碱中、高浓度组细胞存活率和
细胞培养液中Hyp、Col-Ⅰ、Col-Ⅲ含量以及细胞中α-SMA、Smad4 mRNA及其蛋白表达水平均显著降低（P＜0.05），细胞中Smad7
mRNA及其蛋白表达水平均显著升高（P＜0.05）；阳性对照组和苦参碱高浓度组细胞中TGF-β1、TβR-Ⅰ、TβR-Ⅱ、Smad3 mRNA及
其蛋白表达水平均显著降低（P＜0.05）。与苦参碱中浓度组比较，苦参碱高浓度组细胞各指标水平改善程度均更显著（P＜
0.05）。结论：苦参碱能抑制乙醛活化的CFSC-8B细胞的增殖和胶原合成，且具有一定的浓度依赖性；其机制可能与调控TGF-β 1/
Smad信号通路的传导有关。
关键词 苦参碱；肝星状细胞；增殖；胶原合成；转化生长因子β1/Smad通路；机制
Study on the Effects and Mechanism of Matrine on Proliferation and Collagen Synthesis of Hepatic
Stellate Cells CFSC-8B Activated by Acetaldehyde
WANG Xiaoli（School of Pharmacy，Qiqihar Medical University，Heilongjiang Qiqihar 161006，China）

ABSTRACT OBJECTIVE：To study the effects of matrine on proliferation and collagen synthesis of rat hepatic stellate cells
CFSC-8B activated by acetaldehyde，and to investigate its possible mechanism. METHODS：CFSC-8B cells cultured in vitro were
divided into blank control group，model group，positive control group（2.5 μmol/L colchicine）and matrine low，medium and high
concentration groups （30，60，120 μ mol/L）. Except for blank control group，other groups were activated with 200 μ mol/L
acetaldehyde for 24 h；medicine groups were intervened with relevant medicine for 24 h（blank control group and model group
were intervened with equal volume blank medium）. Survival rate of cell was detected by CCK-8 assay. Cells were divided into
blank control group，model group，positive control group（2.5 μmol/L colchicine），matrine medium and high concentration groups
（60，120 μmol/L），then activated and treated with same method. Hydroxyprolin（Hyp）content in cell culture solution was tested
by enzyme digestion. The contents of Col-Ⅰ and Col-Ⅲ in cell culture solution were determined by ELISA. mRNA expressionss of
α-SMA，TGF-β 1，T β R-І ，T β R- Ⅱ ，Smad3，Smad4 and Smad7 in cells were detected by RT-PCR. The protein expressions of
α-SMA，TGF-β 1，TβR-Ⅰ，TβR- Ⅱ，Smad3，Smad4 and Samd7 in cells were detected by Western blotting. RESULTS：Compared
with blank control group，survival rate of cells in model group was increased significantly（P＜0.05）；the contents of Hyp，Col-Ⅰ
and Col-Ⅲ in cell culture solution，mRNA and its protein expressions of α-SMA，TGF-β 1，TβR-Ⅰ，TβR-Ⅱ，Smad3，Smad4 in
cells were increased significantly in model group（P＜0.05），while the mRNA and protein expression of Smad7 was decreased
significantly（P＜0.05）. Compared with model group，survival rate of cells，the contents of Hyp，Col-Ⅰ and Col-Ⅲ in cell culture
solution，the mRNA and protein expressions of α-SMA and Smad4 were decreased significantly in positive control group and
matrine medium and high concentration groups（P＜0.05），
Δ 基金项目：黑龙江省自然科学基金面上项目（No.H2016098）；
黑龙江省省属高等学校基本科研业务费科研项目（No.2018-KYY- while the mRNA and protein expression of Smad7 was
WF-0099） increased significantly （P＜0.05）； the mRNA and protein
＊副教授，博士。研究方向：中药的药理作用及其作用机制。电 expressions of TGF-β 1，TβR-Ⅰ，TβR-Ⅱ and Smad3 were
话：0452-2664571。E-mail：ccwxl@126.com decreased significantly in positive control group and matrine

中国药房 2020年第31卷第11期 China Pharmacy 2020 Vol. 31 No. 11 ·1353 ·