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目的:研究荔枝核总黄酮(TFL)在体外对转化生长因子-β1(TGF-β1)活化的大鼠肝星状细胞(HSC-T6)增殖、凋亡及TLR4/NF-κB途径的影响,探究该药抗肝纤维化的作用机制。方法:体外培养HSC-T6,用TGF-β1刺激24h后以不同浓度的TFL干预24、48、72h,用MTT法检测TFL对HSC-T6增殖的影响,Hoechst 33258荧光染色观察凋亡细胞形态学变化,采用流式细胞术、荧光定量PCR检测TFL对细胞凋亡及α-SMA、TLR4、NF-κB、Survivin m RNA表达水平的影响。结果:TFL以时间和浓度依赖性抑制活化的HSC-T6增殖,并以作用48h最为显著。TFL可促进细胞凋亡,下调HSC-T6中α-SMA、TLR4、NF-кB和Survivin m RNA表达水平,并具有浓度依赖性。同时Hoechst 33258染色显示TFL各组细胞可见细胞核固缩、边聚和裂解、凋亡小体形成等凋亡形态学变化。结论:TFL在体外可以抑制活化的HSC-T6增殖,促进细胞凋亡,其机制可能与降低活化的HSC-T6内TLR4、NF-кB和Survivin m RNA的表达有关。
Objective: To study the effects of total flavonoids of litchi core (TFL) on proliferation, apoptosis and TLR4 / NF-κB pathway in rat hepatic stellate cells (HSC-T6) activated by transforming growth factor-β1 (TGF- Explore the mechanism of action of anti-liver fibrosis. Methods: The HSC-T6 cells were cultured in vitro and stimulated with TGF-β1 for 24 h and 48 h with different concentrations of TFL. MTT assay was used to detect the effect of TFL on HSC-T6 proliferation. Hoechst 33258 staining was used to observe the morphological changes of apoptotic cells The effect of TFL on the apoptosis and the expression of α-SMA, TLR4, NF-κB and Survivin m RNA were examined by flow cytometry and real-time PCR. Results: TFL inhibited the proliferation of activated HSC-T6 in a time-and-concentration-dependent manner, with the most significant effect being at 48 h. TFL can promote apoptosis, down-regulate the expression of α-SMA, TLR4, NF-κB and Survivin m RNA in HSC-T6 in a concentration-dependent manner. At the same time Hoechst 33258 staining showed that all groups of TFL cells showed nuclear morphological changes such as nuclear condensation, edge aggregation and lysis, apoptotic body formation. CONCLUSION: TFL can inhibit the proliferation of HSC-T6 cells and promote the apoptosis of HSC-T6 in vitro. The mechanism may be related to the decrease of the expression of TLR4, NF-κB and Survivin m RNA in activated HSC-T6.