大鼠肝星状细胞CTGF、纤维连接蛋白和整合素的表达变化及肝复康的调节作用

Changes of CTGF,fibronectin and integrin mRNA levels in Gan-fu-kang,a herbal medicine-intervened rat hepatic stellate cells in vitro

目的观察乙醛激活的大鼠肝星状细胞(HSCs)结缔组织生长因子(CTGF)、纤维连接蛋白(FN)和整合素的表达变化,以及中药肝复康对它们表达的影响。方法制备肝复康含药血清,体外培养HSC-T6细胞,用乙醛及肝复康含药血清干预HSCs,采用RT-PCR法检测HSCs中CTGF、FN、整合素α5、整合素β1和α-平滑肌肌动蛋白(α-SMA)m RNA的相对光密度值。结果乙醛刺激后HSC-T6细胞CTGF、FN、整合素α5、整合素β1和α-SMA m RNA水平分别为(1.06±0.02)、(1.28±0.23)、(2.13±0.09)、(1.57±0.50)和(2.84±0.14),与空白对照组细胞[分别为(0.46±0.05)、(0.40±0.09)、(0.37±0.10)、(0.19±0.03)和(0.18±0.03)]相比,均显著升高(P<0.01),而肝复康干预能显著降低它们的水平[分别为(0.59±0.02)、(0.71±0.15)、(1.55±0.04)、(0.92±0.22)和(1.99±0.10),P<0.01]。结论肝复康干预HSCs活化可能与抑制CTGF、FN、整合素α5、整合素β1和α-SMA水平有关。

Objective To observe the changes of connective tissue growth factor （CTGF）,fibronectin （FN）, and integrin mRNA levels in a herbal medicine-intervened HSC-T6 cells in vitro. Methods Sera containing Gan-fu-kang,a herbal medicine compound,was prepared from rats. HSC-T6 cells were cultured in vitro,and the HSCs activated by acetaldehyde were incubated with sera containing Gan-fu-kang. RT-PCR was used to detect the mRNA of CTGF,FN,integrin-α5,integrin-β1 and α-smooth muscle actin （α-SMA）. Result The mRNA levels of CTGF,FN,integrin-α5,integrin-β1,and α-SMA in the HSCs stimulated by acetaldehyde were （1.06± 0.02）, （1.28±0.23）, （2.13±0.09）（1.57±0.50） and （2.84±0.14）,respectively,significantly higher than those in the control cells [（0.46±0.05）, （0.40±0.09）, （0.37±0.10）, （0.19±0.03） and （0.18±0.03）,P〈0.013,and they were down- regulated after incubated with sera containing Gan-fu-kang [（0.59±0.02）,（0.71±0.15）, （1.55±0.04）,（0.92±0.22） and （1.99±0.10）,respectively,P〈0.01]. Conclusion The effect of Gan-fu-kang is related to the inhibition of CTGF, FN,integrin ct5, integrin-β1 and α-SMA in activated HSCs.