目的 研究通過(guò)制備納米脂質(zhì)體提高苦參堿（matrine，MT）去活化肝星狀細(xì)胞的體外作用效果。方法 制備不同載藥量（5%、10%、20%）的苦參堿納米脂質(zhì)體（Nano-MT）并進(jìn)行表征；分析藥物體外釋放行為；給予LX-2細(xì)胞不同濃度的Nano-MT（350.00、175.00、87.50、43.75、21.80、10.90、5.40、2.70、1.40 mmol/L），MTT法檢測(cè)細(xì)胞存活率，計(jì)算半數(shù)抑制濃度（IC₅₀）；考察MT、Nano-MT（10 mmol/L）對(duì)人肝臟星狀細(xì)胞LX-2的去活化效果，細(xì)胞組化染色檢測(cè)平滑肌肌動(dòng)蛋白（α-SMA）、轉(zhuǎn)化生長(zhǎng)因子-β（TGF-β）表達(dá)；實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測(cè)α-SMA、TGF-β、成纖維細(xì)胞生長(zhǎng)因子（FGF）、成纖維細(xì)胞活化蛋白（FAP）mRNA表達(dá)。結(jié)果 Nano-MT透射電鏡表征下呈圓形顆粒狀；載藥量10%制劑載藥量較高，粒徑分布均一，穩(wěn)定性好，滿足給藥需求；制劑中藥物釋放符合Higuchi方程（R²=0.951 3），72 h左右藥物釋放量趨于飽和，累計(jì)釋放量為92.3%。與對(duì)照組和MT組比較，Nano-MT組α-SMA、TGF-β陽(yáng)性細(xì)胞數(shù)均顯著下降（P<0.01、0.001），α-SMA、TGF-β、FGF、FAP mRNA水平顯著降低（P<0.01、0.001）。結(jié)論 Nano-MT具有一定的緩釋作用；可以通過(guò)抑制α-SMA表達(dá)去活化LX-2細(xì)胞抑制肝纖維化。

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

國(guó)家中醫(yī)藥管理局“中醫(yī)藥防治重大疑難疾病臨床服務(wù)能力建設(shè)項(xiàng)目”（國(guó)中醫(yī)藥辦規(guī)財(cái)發(fā)［2013］ 41號(hào)）；湖南省高層次衛(wèi)生人才“225”工程人才項(xiàng)目（湘衛(wèi)函〔2019〕 196號(hào)）；湖南中醫(yī)藥大學(xué)藥學(xué)學(xué)科開放基金（2018YX05）；湖南中醫(yī)藥大學(xué)校級(jí)科研基金項(xiàng)目（2019XJJJ033）