Meng J, Sakata N, Imanaga Y, Tachikawa Y, Chihara J, Takebayashi S
The Second Department of Pathology, Fukuoka University Medical School, Japan.
Clin Nephrol. 1999 May;51(5):280-9.
The purpose of this study is to examine whether or not there is a relationship between glycoxidation and lipid peroxidation in patients with chronic renal failure.
Dermal samples from 26 living or autopsied subjects were sequentially extracted with NaCl, pepsin, collagenase, and NaOH to obtain four fractions (salt-soluble fraction: SSF; pepsin-soluble fraction: PSF; collagenase-soluble fraction: CSF; and insoluble fraction: ISF). The glycoxidation product was measured by pentosidine-linked fluorescence (ex: 335/em: 385) and the levels of lipid peroxide, malondialdehyde (MDA), were assessed by determining the MDA-linked fluorescence (ex: 390/em: 460) which was further confirmed by HPLC.
In patients undergoing hemodialysis, MDA-linked fluorescence markedly increased in collagen-rich fractions, PSF, CSF, and ISF, while pentosidine-linked fluorescence increased in PSF and CSF, in comparison to the controls and the pre-dialysis patients with CRF. Interestingly, the increase in the lipid peroxides strongly correlated with the level of glycoxidation product in PSF, CSF, and ISF (p < 0.0001 in PSF, CSF; p < 0.01 in ISF). The HPLC data of MDA in the PSF was in good correlation with logistic levels of both MDA- (n = 9, r = 0.738, p = 0.023) and pentosidine-linked fluorescence (n = 9, r = 0.721, p = 0.028). In contrast, in SSF, the collagen-poor fraction (collagen content: less than 3% of the total extracted collagen), the data showed a significant increase in the MDA-linked fluorescence only in the pre-dialysis patients with CRF, but not in the HD patients with no correlation with the glycoxidation products.
These findings suggest that both the lipid peroxidation and glycoxidation increased in close relation to each other in the matrix collagen and thus demonstrate a synergetic contribution to the tissue damage observed in patients with CRF
本研究旨在探讨慢性肾衰竭患者糖基化氧化与脂质过氧化之间是否存在关联。
从26名活体或尸检对象获取皮肤样本,依次用氯化钠、胃蛋白酶、胶原酶和氢氧化钠进行提取,以获得四个组分(盐溶组分:SSF;胃蛋白酶溶组分:PSF;胶原酶溶组分:CSF;不溶组分:ISF)。通过戊糖苷连接荧光法(激发波长:335/发射波长:385)测定糖基化氧化产物,并通过测定丙二醛(MDA)连接荧光法(激发波长:390/发射波长:460)评估脂质过氧化物水平,该方法通过高效液相色谱法进一步确认。
与对照组及未透析的慢性肾衰竭患者相比,接受血液透析的患者中,富含胶原的组分(PSF、CSF和ISF)中与MDA相关的荧光显著增加,而PSF和CSF中与戊糖苷相关的荧光增加。有趣的是,脂质过氧化物的增加与PSF、CSF和ISF中糖基化氧化产物水平密切相关(PSF、CSF中p < 0.0001;ISF中p < 0.01)。PSF中MDA的高效液相色谱数据与MDA相关荧光水平(n = 9,r = 0.738,p = 0.023)和戊糖苷连接荧光水平(n = 9,r = 0.721,p = 0.028)均具有良好相关性。相比之下,在SSF(胶原含量低的组分,占总提取胶原的比例小于3%)中,数据显示仅未透析的慢性肾衰竭患者中与MDA相关的荧光显著增加,而血液透析患者中未增加,且与糖基化氧化产物无相关性。
这些发现表明,基质胶原中脂质过氧化和糖基化氧化均相互密切增加,从而证明它们对慢性肾衰竭患者观察到的组织损伤具有协同作用。