南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2014年
6期
837-842
,共6页
殷锦锦%唐外姣%曾璐%周本杰
慇錦錦%唐外姣%曾璐%週本傑
은금금%당외교%증로%주본걸
脂肪肝%脂肪变性%L-02细胞%细胞模型
脂肪肝%脂肪變性%L-02細胞%細胞模型
지방간%지방변성%L-02세포%세포모형
fatty liver%steatosis%L-02 cells%cell model%Huganqingzhi tablet
目的：探讨建立人肝细胞L-02单纯肝脂肪变性细胞模型的方法与模型的应用。方法体外用1640培养基培养L-02细胞，分为模型组和正常组。当细胞融合达到70%~80%，正常组换用新鲜1640培养液，而模型组改为含有游离脂肪酸（油酸和棕榈酸）混合物的1640培养基诱导培养24 h。以油红O染色定性观察细胞内脂滴并通过显色比较各组间总脂质差别，模型鉴定采用观察细胞内脂滴形态，检测细胞内甘油三酯和细胞培养上清丙二醛含量，并检测细胞凋亡情况；同时利用此模型观察护肝清脂片对脂肪变性肝细胞内脂滴的影响。结果模型组L-02细胞内脂滴大量形成，且甘油三酯明显升高，而培养液中肝酶释放量及丙二醛较正常组没有明显升高，且凋亡率没有明显增加；护肝清脂片含药血清两次给药，可以使细胞内脂滴和甘油三酯明显减少。结论采用油酸和棕榈酸混合物（油酸∶棕榈酸=2∶1）可以成功诱导L-02细胞脂肪变性，该模型适用于治疗脂肪肝药物的研究。
目的：探討建立人肝細胞L-02單純肝脂肪變性細胞模型的方法與模型的應用。方法體外用1640培養基培養L-02細胞，分為模型組和正常組。噹細胞融閤達到70%~80%，正常組換用新鮮1640培養液，而模型組改為含有遊離脂肪痠（油痠和棕櫚痠）混閤物的1640培養基誘導培養24 h。以油紅O染色定性觀察細胞內脂滴併通過顯色比較各組間總脂質差彆，模型鑒定採用觀察細胞內脂滴形態，檢測細胞內甘油三酯和細胞培養上清丙二醛含量，併檢測細胞凋亡情況；同時利用此模型觀察護肝清脂片對脂肪變性肝細胞內脂滴的影響。結果模型組L-02細胞內脂滴大量形成，且甘油三酯明顯升高，而培養液中肝酶釋放量及丙二醛較正常組沒有明顯升高，且凋亡率沒有明顯增加；護肝清脂片含藥血清兩次給藥，可以使細胞內脂滴和甘油三酯明顯減少。結論採用油痠和棕櫚痠混閤物（油痠∶棕櫚痠=2∶1）可以成功誘導L-02細胞脂肪變性，該模型適用于治療脂肪肝藥物的研究。
목적：탐토건립인간세포L-02단순간지방변성세포모형적방법여모형적응용。방법체외용1640배양기배양L-02세포，분위모형조화정상조。당세포융합체도70%~80%，정상조환용신선1640배양액，이모형조개위함유유리지방산（유산화종려산）혼합물적1640배양기유도배양24 h。이유홍O염색정성관찰세포내지적병통과현색비교각조간총지질차별，모형감정채용관찰세포내지적형태，검측세포내감유삼지화세포배양상청병이철함량，병검측세포조망정황；동시이용차모형관찰호간청지편대지방변성간세포내지적적영향。결과모형조L-02세포내지적대량형성，차감유삼지명현승고，이배양액중간매석방량급병이철교정상조몰유명현승고，차조망솔몰유명현증가；호간청지편함약혈청량차급약，가이사세포내지적화감유삼지명현감소。결론채용유산화종려산혼합물（유산∶종려산=2∶1）가이성공유도L-02세포지방변성，해모형괄용우치료지방간약물적연구。
Objective To establish an in vitro cell model for investigating hepatic steatosis in non-alcoholic fatty liver disease. Methods L-02 cells cultured in 1640 containing 10%fetal bovine serum were divided into control group and model group. At 70%-80%confluency, L-02 cells in the model group were exposed to a long-chain mixture of free fatty acids (FFA, oleate and palmitate ) for 24 h, and cells in control group were treated with fresh medium. Lipid droplets in the cells were observed and total lipid content was determined with Oil Red O staining. The morphology of lipid droplets, trilyceride level, malonaldehyde content and cell apoptosis rate were evaluated to verify the cell model, and the effect of Huganqingzhi tablet on the lipid droplets was observed. Results A large number of lipid droplets were found in the cell model, which showed markedly increased level of triglyceride without significant changes of malonadehyde content or cell apoptosis rate. Intervention with two doses of Huganqingzhi tablet significantly decreased the number of lipid droplets and trilyceride content in the cell model. Conclusion hepatic steatosis L-02 cell model can be established by long-chain mixture of free fatty acids (oleate∶spalmitate=2∶1) for therapeutic drug studies.
