INFLUENCE OF LIPOPROTEINS ON RENAL CYTOTOXICITY AND ANTIFUNGAL ACTIVITY OF AMPHOTERICIN-B
INFLUENCE OF LIPOPROTEINS ON RENAL CYTOTOXICITY AND ANTIFUNGAL ACTIVITY OF AMPHOTERICIN-B
复制标题
DOI：
10.1128/aac.38.2.223
复制
发表时间：
1994-02-01
影响因子：
4.9
通讯作者：
LOPEZBERESTEIN, G
中科院分区：
文献类型：
--
作者：
WASAN, KM;ROSENBLUM, MG;LOPEZBERESTEIN, G
我们研究了高密度脂蛋白（HDL）和低密度脂蛋白（LDL）对阿朴霉素B（Amp B）对真菌和肾细胞毒性的影响。将白色念珠菌与AmpB和脱氧胆酸盐（Fungizone）或脂质体AmpB（L-AmpB）（0.1至2.0 μ g AmpB/ml）在存在或不存在HDL或LDL（0.5mg蛋白/ml）的情况下在37 ℃下孵育18小时。无论是否存在HDL或LDL，AmpB和L-AmpB的MIC相似。将来源于猪近端肾小管细胞的原代培养物的LLC PK 1肾细胞在无血清培养基中于37 ℃孵育18小时，所述无血清培养基含有AmpB和脱氧胆酸盐或L-AmpB（20 μ g AmpB/ml）、HDL或LDL（0.5mg蛋白质/ml）、AmpB与HDL或LDL的混合物以及L-AmpB与HDL或LDL的混合物。HDL相关AmpB对LLC PK 1细胞的毒性低于AmB（53.0% +/- 2.5% vs 81.3% +/- 3.6%细胞毒性; P = 0.01），而LDL相关AmpB与AmpB毒性相同。L-AmpB、HDL相关L-AmpB和LDL相关L-AmpB对LLC PK 1细胞的毒性低于AmpB（48.3% +/-1.5%、25.5% +/-2.2%和52.2% +/- 2.5% vs 81.3% +/- 3.6%细胞毒性; P = 0.02）。为了进一步理解HDL相关AmpB降低肾细胞毒性作用的原因，检查LLC PK 1细胞中HDL和LDL受体的存在。LLC PK 1细胞表达高亲和力（Kd = 0.0538 ng/ml; 96，000个位点/细胞）和低亲和力（Kd = 222.22 ng/ml; 77个位点/细胞）LDL受体，但仅表达低亲和力HDL受体（Kd = 71.43 ng/ml; 2个位点/细胞）。HDL相关AmpB和LDL相关AmpB对胰蛋白酶化LLC PK 1细胞的毒性低于AmpB（46.6% +/- 10.9%和16.8% +/- 15.98% vs 74.7% +/- 7.7%细胞毒性; P = 0.02）。HDL相关AmB和LDL相关L-AmpB对细胞的毒性也低于AmpB（20.4% +/- 6.2%和13.5% +/- 8.6% vs 74.7% +/- 7.7%细胞毒性; P = 0.01）。AmpB和L-AmpB的抗真菌活性在HDL或LDL存在下没有改变。我们的结论是，降低肾毒性与使用L-AmpB相关的是一个肾细胞的AmpB的摄取减少时，AmpB与HDL，因为在这些细胞中的HDL受体的表达水平低。
We examined the influence of high-density lipoproteins (HDLs) and low-density lipoproteins (LDLs) on the toxicity of amphotericin B (AmpB) to fungal and renal cells. Candida albicans was incubated for 18 h at 37 degrees C with AmpB and deoxycholate (Fungizone) or liposomal AmpB (L-AmpB) (0.1 to 2.0 mu g of AmpB per mi) in the presence or absence of HDLs or LDLs (0.5 mg of protein per mi). The MICs of AmpB and L-AmpB, whether or not HDLs or LDLs were present, were similar. LLC PK1 renal cells, derived from primary cultures of pig proximal tubular cells, were incubated for 18 h at 37 degrees C in serum-free medium that contained AmpB and deoxycholate or L-AmpB at 20 mu g of AmpB per mi, HDLs or LDLs at 0.5 mg of protein per mi, mixtures of AmpB with HDLs or LDLs, and mixtures of L-AmpB with HDLs or LDLs. HDL-associated AmpB was less toxic than AmB to LLC PK1 cells (53.0% +/- 2.5% versus 81.3% +/- 3.6% cytotoxicity; P = 0.01), while LDL-associated AmpB was as toxic as AmpB. L-AmpB, HDL-associated L-AmpB, and LDL-associated L-AmpB were less toxic to LLC PK1 cells than was AmpB (48.3% +/- 1.5%, 25.5% +/- 2.2%, and 52.2% +/- 2.5% versus 81.3% +/- 3.6% cytotoxicity; P = 0.02). To further understand why HDL-associated AmpB reduced renal cytotoxic effects, the LLC PK1 cells were examined for the presence of HDL and LDL receptors. LLC PK1 cells expressed high-affinity (K-d = 0.0538 ng/ml; 96,000 sites per cell) and lo rv-affinity (K-d = 222.22 ng/ml; 77 sites per cell) LDL receptors but only a low-affinity HDL receptor (K-d = 71.43 ng/ml; 2 sites per cell). HDL-associated AmpB and LDL-associated AmpB were less toxic than AmpB to trypsinized LLC PK1 cells (46.6% +/- 10.9% and 16.8% +/- 15.98% versus 74.7% +/- 7.7% cytotoxicity; P = 0.02). HDL-associated AmB and LDL-associated L-AmpB were also less toxic than AmpB to the cells (20.4% +/- 6.2% and 13.5% +/- 8.6% versus 74.7% +/- 7.7% cytotoxicity; P = 0.01). The antifungal activities of AmpB and L-AmpB were not altered in the presence of HDLs or LDLs. We conclude that the reduced nephrotoxicity associated with the use of L-AmpB is related to a decreased uptake of AmpB by renal cells when AmpB is associated with HDLs because of the low level of expression of HDL receptors in these cells.