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目的:观察雷公藤多甙(TW)对肾小球系膜细胞增殖、凋亡、凋亡相关基因 ICE 和 bcl-2表达的影响。方法:采用肾小球系膜细胞培养方法进行体外研究,用不同剂量含雷公藤多甙的大鼠血清刺激4～6代系膜细胞16 h,MTT 法测定系膜细胞增殖指数:TUNEL 法定量观察凋亡细胞;琼脂糖凝胶电泳法观察凋亡细胞核小体 DNA 的断裂现象;逆转录-聚合酶链反应技术(RT-PCR)检测凋亡相关基因 ICE 和 bcl-2的表达变化。结果:加入雷公藤多甙血清16 h 后,系膜细胞出现了典型的凋亡征象,诸如细胞内深棕色颗粒形成、核固缩、碎裂和 DNA 梯形条带等;凋亡率显著高于空白血清组(P<0.01);同时 TW 各组 ICE 表达亦明显高于空白血清对照组,而抑制凋亡的 bcl-2基因各组均表达不明显。结论:雷公藤多甙可诱导系膜细胞凋亡,促进肾小球增殖性病变消散,并可能通过调控 ICE 和 bcl-2基因影响凋亡。
OBJECTIVE: To observe the effects of tripterygium wilfordii (TW) on the proliferation, apoptosis, apoptosis-related gene ICE and bcl-2 expression of glomerular mesangial cells. METHODS: Mesangial cell culture method was used for in vitro studies. Mesangial cells were stimulated with different doses of rat serum containing Tripterygium wilfordii for 16 to 16 hours. The proliferation index of mesangial cells was measured by MTT method: quantification by TUNEL method. The apoptotic cells were observed. Apoptotic nucleosome DNA fragmentation was observed by agarose gel electrophoresis. The expression of apoptosis-related genes ICE and bcl-2 were detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: After the addition of serum of Tripterygium wilfordii for more than 16 h, typical signs of apoptosis appeared in mesangial cells, such as the formation of dark brown particles, nuclear condensation, fragmentation, and DNA trapezoidal bands; the apoptosis rate was significantly higher than that of mesangial cells. In the blank serum group (P<0.01), the expression of ICE in each TW group was also significantly higher than that in the blank serum control group, while the bcl-2 gene that inhibited apoptosis was not significantly expressed in each group. CONCLUSION: Tripterygium wilfordii can induce apoptosis of mesangial cells, promote glomerular proliferative lesions to dissipate, and may affect apoptosis through regulating ICE and bcl-2 genes.