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目的 :观察低密度脂蛋白 (L DL )对体外培养的大鼠肾小球系膜细胞 (Ms C)生长及转化生长因子β(TGF-β)和纤维连接蛋白 (FN)基因表达的影响。 方法 :体外培养的 Ms C培养液中加入 L DL 共同孵育 ,采用 3H- Td R渗入法检测 Ms C增殖情况 ;应用 Northern blot检测 TGF- β m RNA和 FN m RNA的表达 ;应用斑点杂交法检测抗 TGF- β抗体对 FN m RNA表达的影响。结果 :(1) L DL 刺激 Ms C在小剂量以剂量依赖 (5 0～ 15 0 μg/ ml)和时间依赖 (2 4～ 72 h)促进增殖 (P<0 .0 5 ) ,大剂量 (2 0 0μg/ ml)抑制增殖 (P<0 .0 5 )。 (2 ) Northern blot显示 L DL 刺激 Ms C增殖 ,其 TGF- β和 FN m RNA表达以剂量和时间依赖方式增强。 TGF-β m RNA表达较 FN m RNA提前 2 4h。 (3) Dot blot显示 ,加入抗 TGF-β抗体后 ,FN m RNA的表达较未加前明显减弱。 结论 :L DL不仅可刺激肾小球 Ms C的增殖 ,并且增加 TGF-β和 FN m RNA的表达。
Objective: To observe the effect of low density lipoprotein (L DL) on the growth of rat mesangial cells (Ms C) and the expression of transforming growth factor β (TGF-β) and fibronectin (FN) genes in vitro. Methods: MSCs cultured in vitro were incubated with L DL, and the proliferation of Ms C was detected by 3H-TdR infiltration method. The expression of TGF-β m RNA and FN m RNA was detected by Northern blot. The dot blot hybridization assay Effect of Anti-TGF-β Antibody on FN m RNA Expression. Results: (1) Ms DL stimulated the proliferation of Ms C in a dose-dependent manner (50 ~ 150 μg / ml) and time-dependent manner (24 ~ 72 hours) 200 μg / ml) inhibited proliferation (P <0.05). (2) Northern blot showed that L DL stimulated the proliferation of Ms C, and the expression of TGF-β and FN m RNA increased in a dose-and time-dependent manner. TGF-β m RNA expression 24 h earlier than FN m RNA. (3) Dot blot showed that the expression of FN m RNA was significantly decreased after anti-TGF-β antibody was added. CONCLUSION: L-DL not only stimulates the proliferation of glomerulus Ms C, but also increases the expression of TGF-β and FN m RNA.