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[摘要]
目的 探究胡黃連苷Ⅰ、Ⅱ、Ⅲ、Ⅳ體外抗非酒精性脂肪性肝炎(NASH)的活性。方法 用1 mmol/L游離脂肪酸(FFA)誘導HepG2細胞24 h,建立體外NASH模型;MTT法檢測胡黃連苷Ⅰ、Ⅱ、Ⅲ、Ⅳ對細胞活力的影響,并依此優(yōu)選3個安全給藥濃度(0.25、0.50、1.00 mg/mL)干預體外NASH模型;油紅O染色法鏡下觀察細胞內(nèi)脂滴的變化情況;試劑盒法檢測細胞培養(yǎng)上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-8(IL-8)、腫瘤壞死因子-α(TNF-α)的含量變化。結果 胡黃連苷Ⅰ、Ⅱ、Ⅲ、Ⅳ對HepG2細胞的活力影響較小。FFA刺激24 h后,模型組與對照組比較,細胞內(nèi)被染成明顯的紅色顆粒狀脂滴,細胞培養(yǎng)上清液中SOD水平顯著下降,MDA、TNF-α、IL-8水平顯著上升,均有顯著性差異(P<0.05、0.01)。與模型組比較,不同濃度的胡黃連苷Ⅰ、Ⅱ、Ⅲ、Ⅳ干預組,細胞內(nèi)紅色顆粒狀脂滴減少,且以胡黃連苷Ⅱ最為明顯;SOD水平顯著上升,MDA、TNF-α、IL-8水平顯著下降(P<0.05、0.01),且以胡黃連苷Ⅱ、Ⅲ、Ⅳ較為明顯,但沒有顯著的劑量相關性。結論 胡黃連苷Ⅰ、Ⅱ、Ⅲ、Ⅳ中胡黃連苷Ⅱ的體外抗NASH的活性較顯著。
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[Abstract]
Objective To discuss the anti-NASH activity of PicrosideⅠ, PicrosideⅡ, PicrosideⅢ and PicrosideⅣ in vitro.Methods Liver cells HepG2 were cultured and induced by FFA to establish NASH cell model. The cell viability were measured by MTT to select the safe drug concentration. Then safe concentrations of PicrosideⅠ, PicrosideⅡ, PicrosideⅢ and PicrosideⅣ were meanwhile added. The fat deposition in cells was observed by oil-red O staining method. The content of SOD, MDA, IL-8 and TNF-α were tested by Elisa. Results Three treatment groups were seted with the concentration of 0.25, 0.50 and 1.00 mg/mL. Compared with control group, the NASH cell model group was stained with obvious red granular lipid droplets. The levels of SOD were significantly higher in model group than the control group, while MDA, TNF-α and IL-8 significantly lower than control group (P<0.05 and 0.01). Compared with the model group, the red granular lipid droplets decreased in the intervention groups of different concentrations of Picroside I, Ⅱ, Ⅲ and IV, especially Picroside Ⅱ. SOD levels increased significantly, MDA, TNF-a and IL-8 levels decreased significantly (P<0.05, 0.01), and Picroside Ⅱ, Ⅲ and IV were more obvious, but there was no significant dose correlation. Conclusion PicrosideⅡcan improve cellular pathology and influence the levels of SOD, MDA, IL-8 and TNF-α obviously, showing potential value on NASH treatment.
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