山萘酚對HepG2細胞凋亡的影響

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1.山西醫科大學附屬第二臨床醫院消化內科，山西 太原 030001；2.首都醫科大學附屬北京佑安醫院重癥肝病科；3.首都醫科大學附屬北京佑安醫院北京市肝病研究所；4.山西省腫瘤醫院 山西省腫瘤研究所

【Abstract】ObjectiveTo investigate the effect of kaempferol on the apoptosis of human hepatocellular carcinoma HepG2 cells.MethodsHepG2 cells were incubated with different concentrations of kaempferol for 24 hours to select the best drug concentration (100 μmol/L), and HepG2 cells were incubated at different times. Cell viability was measured by MTT assay; lactate dehydrogenase (LDH) activity in cell supernatants was measured by LDH assay kit; apoptosis was measured by flow cytometry. The expression of apoptosis-related protein was detected by Western blotting.ResultsKaempferol had the induced effect on apoptosis of HepG2 cells in a dose- and time-dependent. When HepG2 cells were treated with kaempferol for 24 hours, the cells viability was gradually decreased and the apoptotic rate was increased gradually with the increase of kaempferol concentrations. Compared with the normal control group, in the 100 μmol/L kaempferol for 24 hours group, the cells survival rate was significantly decreased [(60.58±3.74)%vs(100±2.28)%,P<0.0001], and the apoptotic rate was significantly increased [(16.43±0.07)%vs(0.68±0.09)%,P<0.0001]. The expression of apoptotic factor Cleaved-caspase3 protein was increased with the increase of kaempferol concentrations， HepG2 cells were incubated with kaempferol for 24 hours by Western blotting. When the kaempferol concentration was 100 μmol/L, the apoptotic rate also was increased gradually with the prolongation of time when HepG2

cells were incubated with kaempferol. Compared with the normal control group, in the 100 μmol/L kaempferol for 24 hours group, the cell survival rate was significantly decreased [(59.36±3.09)%vs(100±2.28)%,P<0.0001], and the apoptotic rate was significantly increased [(16.71±1.12)%vs(0.35±0.01)%,P<0.0001]. The expression of apoptotic factor Cleaved-caspase3 was increased with the prolongation of time.ConclusionKaempferol has an induced effect on the apoptosis of human hepatocellular carcinoma cell line HepG2 cells in dose- and time-dependent. It indicats that high concentration of kaempferol could induce the apoptosis of HepG2 cells by promoting the expression of apoptotic factor Cleaved-caspase3, and thus it has the potential effect on anti-hepatocarcinoma activity.

【Keywords】 Kaempferol； HepG2 cells； Cell apoptosis； Mechanism

山萘酚是一種多酚類抗氧化物，為天然黃酮類化合物的一種(化學結構見圖1)，其主要來源于姜科植物山萘的根莖，中草藥淫羊藿、石菖蒲、苦參、山豆根等均含有此成分，同時廣泛存在于日常食用的蔬菜、水果當中。山萘酚具有抗氧化、抗炎、抗菌、抗癌、保肝等功效。據研究證實，山萘酚可通過阻礙NF-κB信號通路來抑制炎癥因子IL-6、TNF-α的表達，可對因為炎癥因子誘發的哮喘、肝細胞炎癥等發揮預防及治療作用；山萘酚具有激活過氧化物酶增殖物激活受體(PPAR-γ)的作用，從而提高胰島素敏感性，增強葡萄糖儲存能力，其中PPAR-γ激動劑已成為脂質代謝異常和2型糖尿病的常用藥物，據此推測山萘酚將可能成為治療糖尿病的臨床用藥；山萘酚還可提高細胞氧化應激能力，降低活性氧的產生，有顯著的抗氧化活性，具有抗心肌細胞缺氧復氧損傷作用。山萘酚不僅有如上作用，還有誘導癌細胞凋亡的功效，據相關研究[5-8]表明，山萘酚有抗胃癌、結腸癌、胰腺癌、腎癌等功能，然而關于山萘酚對肝癌的研究為數甚少且相關機制說法不一。所以本次研究通過體外實驗，來探討山萘酚對于人類肝癌細胞HepG2凋亡的影響，從而為日后山萘酚用于藥品研發及肝癌的臨床治療提供實驗依據。……