金安粉针剂对肺癌细胞系超微结构的影响

目的:观察中药金安粉针剂对培养的肺癌细胞系超微结构和线粒体形态及功能的影响。方法:将培养的肺癌PG、PAa细胞系分别分为4组:空白对照组(C)、顺铂组(DDP)、金安组(JA)和顺铂合金安组(DJ)。药物作用24h后,采用倒置显微镜、扫描电镜、透射电镜观察其形态变化,并观察药物作用细胞24h、48h,分别用rhodamine、BCECF、Fluo-3染色,流式细胞仪检测线粒体膜电位、细胞内pH和Ca²⁺浓度变化。结果:在倒置显微镜下,各用药组PG、PAa细胞均可见细胞体积缩小、变圆以及脱壁现象,其中DDP组、DJ组尤为明显;扫描电镜见两种瘤细胞用药组细胞多呈现微绒毛减少、融合或断裂,并有细胞穿孔现象;透射电镜观察PG、PAa各组有退行性变,表现为细胞核染色质浓缩、边集或核碎裂。胞浆内可见自噬溶酶体增多,髓样小体多见。线粒体减少并普遍肿胀,其肿胀有两种表现:一种为基质密度增加,膜间隙增宽而线粒体大小变化不大;另一种为空泡变性,基质密度降低。PG用药各组与对照组相比,药物作用24h对线粒体膜电位影响不大,48h则可使线粒体膜电位升高;PAa细胞,24h和48hJA组线粒体膜电位有所下降,其它各组则升高;细胞内Ca²⁺在PG细胞DDP作用24h时升高,而JA、DJ则降低;作用48h时各组均升高;在PAa细胞DDP组24h、48h细胞内Ca²⁺变化不大,而JA、DJ组均明显下降;细胞内pH值在PG、PAa各组药物作用24h均降低,48h则升高。以上PG、PAa各用药组与对照组比均有显著差异,P＜0.05。结论:中药金安及金安合顺铂组引起PG、PAa细胞凋亡和坏死,线粒体有基质密度增加和空泡变性两种改变,而线粒体膜电位、细胞内Ca²⁺、pH有升高的趋势,因此认为线粒体损伤可能是在金安及金安合顺铂导致肺癌细胞凋亡中起重要作用。

Effect of Jinan injection on ultrastructure of lung cancer cell lines

AIM: To investigate the effects of Chinese medicine, Jinan injection, on ultrastructure and mitochondria in cultured lung cancer cell lines. METHODS: The cultured lung cancer cell lines PG and PAa were used and divided into 4 groups: control (C), cisplatin (DDP), Jinan (JA) and Jinan in combination with cisplatin (DJ), respectively. The changes of morphology and mitochondria membrane potential, intracellular Ca~(2 ) and pH in every group were observed by inverted microscope and electronic microscope as well as by using flow cytometry, staining by rhodamine, Fluo-3 and BCECF, respectively. RESULTS: Degeneration cells showed chromatin condensation and peripheral congregation. In cytoplasm autophage lysosome increased and myelinoid body was seen easily. In mitochondria structure, where the space between the inner and outer membranes of these organelles expanded as the matrix was compressed. The electron-dense or swelled was observed as vacuole degeneration and its matrix showed electron-lucent. Compared to control, mitochondria membrane potential increased in every group after 24 h and 48 h treatment. DDP increased intracellular calcium ion in PG cells, however, in PAa cells, JA and DJ decreased it. Intracellular pH got lower at 24 h and higher at 48 h in PG and PAa cells. There were significance in every group vs control in PG and PAa by statistic t-test (P<0.01). CONCLUSION: Apoptosis was induced in PG and PAa cell lines by Jinan injection and DJ. Mitochondria matrix displayed electron-dense, mitochondrial potential, intracellular calcium ion and pH showed an increasing trend. Mitochondria damages may play an important role in apoptosis induced by Jinan and DJ.