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目的:从细胞水平观察百令提取液对环孢素A(CsA)肾毒性的影响。 方法:体外培养肾小管上皮细胞,通过3H-TdR掺入、MTT比色法、PI染色法和细胞LDH释放等观察百令提取液对CsA引起的肾小管上皮细胞毒性的防治作用。 结果:3H-TdR掺入和MTT比色法显示在一定的剂量范围百令提取液可明显促进培养的肾小管上皮细胞增生, 5mg/L以上的CsA抑制细胞增殖;细胞周期的研究显示15mg/L以上的CsA使小管上皮细胞细胞周期阻滞在G2期;此外,更大剂量的CsA(25mg/L)对细胞有直接的毒性作用,使培养上清LDH值升高。百令提取液能部分逆转CsA引起的细胞增殖受抑和细胞周期阻滞,但是对CsA导致 的细胞培养上清LDH的升高没有明显的影响。 结论:本研究证实了百令提取液可以促进体外培养的肾小管上皮细胞增生,对CsA的肾小管细胞毒性具有一定的拮抗作用。
OBJECTIVE: To observe the effect of Bailing extract on cyclosporine A (CsA) nephrotoxicity at the cellular level. METHODS: Renal tubular epithelial cells were cultured in vitro. The effect of Bailing extract on the cytotoxicity of renal tubular epithelial cells induced by CsA was observed by 3H-TdR incorporation, MTT colorimetry, PI staining and cell LDH release. RESULTS: 3H-TdR incorporation and MTT colorimetric assay showed that Bailing extract could significantly promote the proliferation of cultured renal tubular epithelial cells in a certain dose range. CsA above 5mg/L inhibited cell proliferation; cell cycle studies showed that 15mg/L CsA above L causes cell cycle arrest of tubular epithelial cells in G2 phase; in addition, a higher dose of CsA (25 mg/L) has a direct toxic effect on cells and increases the LDH level in the culture supernatant. Bailing extract partially reversed CsA-induced inhibition of cell proliferation and cell cycle arrest, but had no significant effect on CSH-induced LDH elevation in cell culture supernatants. Conclusion: This study confirms that Bailing extract can promote the proliferation of renal tubular epithelial cells cultured in vitro, and has a certain antagonistic effect on the tubular cytotoxicity of CsA.