脂联素抑制大鼠肝星状细胞株HSC-T6的Ⅲ型前胶原和透明质酸的表达

Inhibition effect of adiponectin on the levels of PC m and HA in HSC-T6 cell

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.