目的 观察脂联素对肝星状细胞(HSCs)分泌结缔组织生长因子(CTGF)的影响,并观察Ⅲ型前胶原(PCⅢ)、透明质酸(HA)含量的变化.方法 把大鼠HSCs分成5组,其中4组分别加入不同浓度的脂联素,另外1组作对照.通过RT-PCR和Western Blot技术分析CTGF mRNA和蛋白表达水平,ELISA法检测上清液PCⅢ、HA含量.结果 与对照组比较,A、B、C、D组脂联素HSCs内CTGF mRNA表达均下降,吸光度比值分别为1.54±0.18、1.21±0.14、0.96±0.10、0.79±0.08,其中以D组下降最明显(t=2.42,P<0.05;t=2.73,P<0.05;t=3.28,P<0.01;t=4.67,P<0.01);CTGF蛋白表达也抑制,积分吸光度比值分别为1.54±0.18、1.21±0.14、0.96±0.10、0.79±0.08,其中以D组抑制最明显(t=2.84,P<0.01;t=4.05,P<0.01;t=6.25,P<0.01;t=9.72,P<0.01);上清液PCⅢ、HA含量也下降,随脂联素浓度增加,下降越明显.结论 脂联素能够抑制HSCs分泌PCⅢ、HA;脂联素可通过抑制HSCs内CTGF表达而发挥抗纤维化作用.
Objective To observe the effects of adiponectin on mRNA and protein expressions of connective tissue growth factor (CTGF) in hepatic stellate cells (HSCs) and the levels of procollagen type Ⅲ (PC Ⅲ) and hyaluronic acid (HA).Methods Cultured rat HSCs were treated with different concentrations of adiponectin.CTGF mRNA and protein expressions were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.The levels of PCⅢ and HA were detected by enzymelinked immunosorbent assay (ELISA).Results Compared with the control group,the result of RT-PCR showed that the four groups had different degrees of inhibitory effect,of which group D exhibited the strongest inhibitory effect.The absorbance ratio was 1.54 ±0.18,1.21 ±0.14,0.96 ±0.10,and 0.79 ± 0.08,respectively (t =2.42,P 〈0.05;t =2.73,P 〈0.05;t =3.28,P 〈0.01;t =4.67,P 〈0.01).Western Blot also indicated that four groups had different degrees of inhibitory effect,of which D group exhibited the strongest inhibitory effect.The ratio of integral absorption was 1.54 ± 0.18,1.21 ±0.14,0.96±0.10,and 0.79 ±0.08,respectively (t =2.84,P 〈0.01;t =4.05,P 〈0.01;t =6.25,P 〈0.01;t =9.72,P 〈0.01).The levels of PCⅢ and HA secreted in culture media were also decreased.It was significantly decreased with the concentration of adiponectin increased.Conclusion Adiponectin can inhibit the levels of PC Ⅲ and HA,which may be achieved through reducing CTGF mRNA and protein expressions.