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有机渗透溶质会增加肾细胞中的细胞质粘度。

Organic osmolytes increase cytoplasmic viscosity in kidney cells.

作者信息

Periasamy N, Kao H P, Fushimi K, Verkman A S

机构信息

Department of Medicine, University of California, San Francisco 94143-0532.

出版信息

Am J Physiol. 1992 Oct;263(4 Pt 1):C901-7. doi: 10.1152/ajpcell.1992.263.4.C901.

Abstract

The hypothesis was tested that accumulation of osmolytes by kidney cells grown in hyperosmolar media decreases the rotational and translational mobilities of small polar solutes in the cytosolic compartment. Rotational mobility was measured by the picosecond rotational correlation times (tau c) of 2',7'-bis(2-carboxyethyl)-5(6)carboxylfluorescein (BCECF) by multiharmonic microfluorimetry. In isolated segments of rabbit proximal tubule, thick ascending limb, and cortical collecting duct that were perfused and bathed in 300 mosM media, tau c were in the range 180-250 ps, corresponding to apparent rotational viscosities (eta r) of 1.1-1.5 cP. In cortical collecting tubule, eta r was not influenced by serosal vasopressin. In Madin-Darby canine kidney (MDCK) cells grown in 300-1,200 mosM media, eta r increased progressively by up to a factor of 1.38 +/- 0.03; measurements of tau c and macroscopic viscosity in artificial solutions containing osmolytes supported the hypothesis that the increased eta r was due to accumulation of organic osmolytes. BCECF translational mobility was measured by fluorescence photobleaching recovery using a focused 1.2-microns diameter Ar laser beam at 488 nm. Recovery half-times were 36 +/- 3 (SE) ms (n = 10) in MDCK cells grown in 300 mosM media and 62 +/- 3 ms (n = 10) when grown in 1,200 mosM media. The results suggest that accumulation of osmolytes by renal cells is associated with significantly increased cytosolic viscosity. The increased viscosity would slow enzymatic and transport processes in the cytosolic compartment.

摘要

该假说得到了验证,即在高渗培养基中生长的肾细胞积累渗透溶质会降低胞质区室中小极性溶质的旋转和平动迁移率。通过多谐波显微荧光法,利用2',7'-双(2-羧乙基)-5(6)羧基荧光素(BCECF)的皮秒旋转相关时间(τc)来测量旋转迁移率。在灌注并浸泡于300 mosM培养基中的兔近端小管、髓袢升支粗段和皮质集合管的分离节段中,τc在180 - 250 ps范围内,对应于1.1 - 1.5 cP的表观旋转粘度(ηr)。在皮质集合管中,ηr不受浆膜血管加压素的影响。在300 - 1200 mosM培养基中生长的Madin-Darby犬肾(MDCK)细胞中,ηr逐渐增加,最高可达1.38±0.03倍;在含有渗透溶质的人工溶液中对τc和宏观粘度的测量支持了以下假说,即ηr的增加是由于有机渗透溶质的积累。通过使用聚焦的直径为1.2微米的488 nm氩激光束进行荧光漂白恢复来测量BCECF的平动迁移率。在300 mosM培养基中生长的MDCK细胞中,恢复半衰期为36±3(SE)ms(n = 10),而在1200 mosM培养基中生长时为62±3 ms(n = 10)。结果表明,肾细胞积累渗透溶质与胞质粘度显著增加有关。粘度增加会减慢胞质区室中的酶促和转运过程。

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