目的 研究通过制备纳米脂质体提高苦参碱（matrine，MT）去活化肝星状细胞的体外作用效果。方法 制备不同载药量（5%、10%、20%）的苦参碱纳米脂质体（Nano-MT）并进行表征；分析药物体外释放行为；给予LX-2细胞不同浓度的Nano-MT（350.00、175.00、87.50、43.75、21.80、10.90、5.40、2.70、1.40 mmol/L），MTT法检测细胞存活率，计算半数抑制浓度（IC₅₀）；考察MT、Nano-MT（10 mmol/L）对人肝脏星状细胞LX-2的去活化效果，细胞组化染色检测平滑肌肌动蛋白（α-SMA）、转化生长因子-β（TGF-β）表达；实时荧光定量PCR（qRT-PCR）法检测α-SMA、TGF-β、成纤维细胞生长因子（FGF）、成纤维细胞活化蛋白（FAP）mRNA表达。结果 Nano-MT透射电镜表征下呈圆形颗粒状；载药量10%制剂载药量较高，粒径分布均一，稳定性好，满足给药需求；制剂中药物释放符合Higuchi方程（R²=0.951 3），72 h左右药物释放量趋于饱和，累计释放量为92.3%。与对照组和MT组比较，Nano-MT组α-SMA、TGF-β阳性细胞数均显著下降（P<0.01、0.001），α-SMA、TGF-β、FGF、FAP mRNA水平显著降低（P<0.01、0.001）。结论 Nano-MT具有一定的缓释作用；可以通过抑制α-SMA表达去活化LX-2细胞抑制肝纤维化。

Objective To study the effect of improving matrine (MT) on deactivating hepatic stellate cells and inhibiting liver fibrosis in vitro by nanoliposome delivery technology. Methods Matrine nanoliposomes (nano-MT) with different drug loading (5%, 10%, 20%) were prepared and characterized. The drug release behavior in vitro was analyzed. LX-2 cells were given different concentrations of nano-MT (350.00, 175.00, 87.50, 43.75, 21.80, 10.90, 5.40, 2.70, 1.40 mmol/L), the cell survival rate was detected by MTT method, and the half inhibitory concentration (IC₅₀) was calculated. The expression of α-SMA and transforming growth factor-β (TGF-β) was detected by histochemical staining. The mRNA expression of α -SMA, TGF-β, fibroblast growth factor (FGF) and fibroblast activating protein (FAP) were detected by real-time quantitative PCR (qRT-PCR). Results Matrine loaded nanoliposome are round particles under transmission electron microscope characterization. The drug loading of 10% preparation was high, the particle size distribution was uniform, and the stability was good, which can meet the needs of drug delivery. Drug release profile of the formulation conforms to the Higuchi equation (R²=0.965 13). Drug release tends to 92.3% at time point of 72 h. Compared with control and MT group, the number of α-SMA and TGF-β positive cells in nano-MT group were significantly decreased (P<0.01 and 0.001), and the mRNA levels of α-SMA, TGF-β, FGF and FAP were significantly decreased (P<0.01 and 0.001). Conclusion Martrine loaded liposomes have good pharmaceutical properties and have a certain sustained release effect. The liposome can inhibit liver fibrosis by effectively inhibiting the expression of α -SMA in liver stellate cells, thereby inhibiting liver fibrosis.

R943;R285.5

国家中医药管理局“中医药防治重大疑难疾病临床服务能力建设项目”（国中医药办规财发［2013］ 41号）；湖南省高层次卫生人才“225”工程人才项目（湘卫函〔2019〕 196号）；湖南中医药大学药学学科开放基金（2018YX05）；湖南中医药大学校级科研基金项目（2019XJJJ033）