1. 贵州中医药大学第二临床医学院
2. 贵州中医药大学
纸质出版:2019
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[1]肖政华,邹艳,杨辉,谭芊任,崔峻松,胡芳.扶肝化纤汤含药血清对TGF-β1诱导HSC-T6细胞增殖及TGF-β1/Smad信号通路的影响[J].中医杂志,2019,60(19):1673-1678.
XIAO Zhenghua, ZOU Yan, YANG Hui, et al. Effects of Serum Containing Fugan Huaxian Decoction(扶肝化纤汤) on TGF-β1 Induced HSC-T6 Proliferation and TGF-β 1/Smad Signaling Pathway[J]. Journal of Traditional Chinese Medicine2019, 60(19): 1673-1678.
[1]肖政华,邹艳,杨辉,谭芊任,崔峻松,胡芳.扶肝化纤汤含药血清对TGF-β1诱导HSC-T6细胞增殖及TGF-β1/Smad信号通路的影响[J].中医杂志,2019,60(19):1673-1678. DOI: 10.13288/j.11-2166/r.2019.19.012.
XIAO Zhenghua, ZOU Yan, YANG Hui, et al. Effects of Serum Containing Fugan Huaxian Decoction(扶肝化纤汤) on TGF-β1 Induced HSC-T6 Proliferation and TGF-β 1/Smad Signaling Pathway[J]. Journal of Traditional Chinese Medicine2019, 60(19): 1673-1678. DOI: 10.13288/j.11-2166/r.2019.19.012.
目的探讨扶肝化纤汤抗肝纤维化的可能作用机制。方法将40只雄性Wistar大鼠随机分为正常对照组、扶肝化纤汤组
每组20只
扶肝化纤汤组以16. 78g/kg扶肝化纤汤灌胃
正常对照组予等体积生理盐水灌胃
每日1次
连续7天
末次给药后于大鼠腹主动脉取血制备含药血清。培养大鼠肝星状细胞HSC-T6
设置空白对照组及1%、2%、5%、8%、10%、15%、20%扶肝化纤汤含药血清组
检测不同浓度扶肝化纤汤含药血清对转化生长因子β1 (TGF-β1)诱导的HSC-T6活化增殖的影响
确定15%扶肝化纤汤含药血清为最佳给药浓度。细胞实验分为空白对照组(HSC-T6细胞)、正常组(HSC-T6细胞+正常对照血清+TGF-β1)、中药组(HSC-T6细胞+15%扶肝化纤汤含药血清+TGF-β1);阻断组1 (HSC-T6细胞+SIS3)、阻断组2 (HSC-T6细胞+正常对照血清+TGF-β1+SIS3)及阻断组3 (HSC-T6细胞+15%扶肝化纤汤含药血清+TGF-β1+SIS3)
培养24 h后比较各组HSC-T6细胞增殖抑制率及细胞中Smad 3、Smad 7、α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原mRNA表达。结果与空白对照组比较
正常组、中药组、阻断组2、阻断组3 HSC-T6细胞增殖抑制率差异有统计学意义(P <0. 01)。与正常组比较
中药组、阻断组2、阻断组3HSC-T6细胞增殖抑制率差异均有统计学意义(P <0. 01)。与中药组比较
阻断组3抑制率明显升高(P <0. 01)。与空白对照组比较
中药组、阻断组2、阻断组3均可下调Smad 3、Ⅰ型胶原mRNA表达
上调Smad 7、α-SMA mRNA表达(P <0. 01);正常组可上调Smad 3、α-SMA、Ⅰ型胶原mRNA表达
下调Smad 7mRNA表达(P <0. 01)。与正常组比较
中药组、阻断组2、阻断组3均可下调Smad 3、α-SMA、Ⅰ型胶原mRNA表达
上调Smad 7 mRNA表达(P <0. 01)。与中药组比较
阻断组3可下调Smad 3、α-SMA、Ⅰ型胶原mRNA表达
上调Smad 7 mRNA表达(P <0. 01)。结论扶肝化纤汤能够抑制TGF-β1诱导的HSC-T6细胞的活化增殖
其可能通过影响TGF-β1/Smad信号转导通路发挥抗肝纤维化作用。
Objective To explore the possible mechanism of anti-liver fibrosis in Fugan Huaxian Decoction( 扶肝化纤汤). Methods A total of 40 male Wistar rats were randomly divided into a normal control group and a Fugan Huaxian Decoction group
with 20 rats in each group. Fugan Huaxian Decoction group was intragastrically administered with 16. 78 g/kg Fugan Huaxian Decoction
and the normal control group was given equal volume of normal saline by gavage
once a day for 7 consecutive days. After the last administration
blood was taken from the abdominal aorta of the rats to prepare the drug-containing serum. Rat hepatic stellate cells HSC-T6 were cultured
and blank control group as well as 1%
2%
5%
8%
10%
15% and 20% Fugan Huaxian Decoction containing serum group were set up. The effects of different concentrations of Fugan Huaxian Decoction serum on the activation and proliferation of HSC-T6 induced by transforming growth factor β1( TGF-β1) were determined. The serum containing15% Fugan Huaxian Decoction was determined as the optimal concentration. Cell experiments were divided into a blank control group( HSC-T6 cells)
a normal group( HSC-T6 cells + normal control serum + TGF-β1)
a Chinese medicine group( HSC-T6 cells + 15% Fugan Huaxian Decoction serum + TGF-β1); a blocking group 1( HSC-T6 cells + SIS3)
a blocking group 2( HSC-T6 cells + normal control serum + TGF-β1 + SIS3) and a blocking group 3( HSC-T6 cells + 15% Fugan Huaxian Decoction containing serum + TGF-β1 + SIS3). After 24 hours of culture
the proliferation inhibition rate of HSC-T6 cells and Smad 3
Smad 7
α smooth muscle actin( α-SMA) and type I collagen mRNA of each group were compared. Results Compared with the blank control group
the inhibition rate of HSC-T6 cell proliferation in the normal group
the Chinese medicine group
the blocking group and the blocking group had statistically significant difference( P < 0. 01). Compared with the normal group
the inhibition rate of HSC-T6 cell proliferation in the Chinese medicine group
the blocking group 2 and the blocking group 3 had statistically significant difference( P < 0. 01). Compared with the Chinese medicine group
the inhibition rate of blocking group 3 was significantly increased( P < 0. 01). Compared with the blank control group
the expression of Smad 3 and type I collagen mRNA was down-regulated in the Chinese medicine group
blocking group 2 and blocking group 3
and the expression of Smad 7 and α-SMA mRNA was up-regulated( P < 0. 01). The normal group could upregulate Smad 3 and α-SMA and type I collagen mRNA expression
down-regulated Smad 7 mRNA expression( P <0. 01). Compared with the normal group
the Chinese medicine group
the blocking group 2 and the blocking group3 all down-regulated the expression of Smad 3
α-SMA and type I collagen and up-regulated the expression of Smad 7 mRNA( P < 0. 01). Compared with the Chinese medicine group
blocking group 3 could down-regulate the expression of Smad 3
α-SMA and type I collagen and up-regulate the expression of Smad 7 mRNA( P < 0. 01). Conclusion Fugan Huaxian Decoction could inhibit the activation and proliferation of HSC-T6 cells induced by TGF-β1
which may play an anti-fibrosis effect by affecting TGF-β1/Smad signal transduction pathway.
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