Basic Research Lab-LIM 12, Nephrology-HCFMUSP, São Paulo, São Paulo, Brazil.
PLoS One. 2012;7(6):e37625. doi: 10.1371/journal.pone.0037625. Epub 2012 Jun 6.
Leptospirotic renal lesions frequently produce a polyuric form of acute kidney injury with a urinary concentration defect. Our study investigated a possible effect of the glycolipoprotein, (GLPc) extracted from L. interrogans, on vasopressin (Vp) action in the guinea pig inner medullary collecting duct (IMCD).
The osmotic water permeability (Pf µm/s) was measured by the microperfusion in vitro technique. AQP2 protein abundance was determined by Western Blot. Three groups were established for study as follows: Group I, IMCD from normal (ngp, n = 5) and from leptospirotic guinea-pigs (lgp-infected with L. interrogans serovar Copenhageni, GLPc, n = 5); Group II, IMCD from normal guinea-pigs in the presence of GLPc (GLPc group, n = 54); Group III, IMCD from injected animals with GLPc ip (n = 8).
In Group I, PFS were: ngp--61.8±22.1 and lgp--8.8±12.4, p<0.01 and the urinary osmolalities were: lgp--735±64 mOsm/Kg and ngp--1,632±120 mOsm/Kg. The lgp BUN was higher (176±36 mg%) than the ngp (56±9 mg%). In Group II, the Pf was measured under GLPc (250 µg/ml) applied directly to the bath solution of the microperfused normal guinea-pig IMCDs. GLPc blocked Vp (200 pg/ml, n = 5) action, did not block cAMP (10(-4) M), and Forskolin (Fors--10(-9) M) action, but partially blocked Cholera Toxin (ChT--10(-9) M) action. GLP from L.biflexa serovar patoc (GLPp, non pathogenic, 250 µg) did not alter Vp action. In Group III, GLPc (250 µg) injected intraperitoneally produced a decrease of about 20% in IMCD Aquaporin 2 expression.
The IMCD Pf decrease caused by GLP is evidence, at least in part, towards explaining the urinary concentrating incapacity observed in infected guinea-pigs.
钩端螺旋体肾病变常导致多尿型急性肾损伤和尿浓缩功能缺陷。本研究探讨了从问号钩端螺旋体中提取的糖脂蛋白(GLPc)对豚鼠内髓集合管(IMCD)中血管加压素(Vp)作用的可能影响。
通过体外微灌注技术测量渗透水通透性(Pf µm/s)。通过 Western Blot 测定 AQP2 蛋白丰度。研究建立了三组:组 I,正常(ngp,n=5)和钩端螺旋体感染(感染了哥本哈根血清型的问号钩端螺旋体,GLPc,n=5)的豚鼠的 IMCD;组 II,正常豚鼠在 GLPc 存在下的 IMCD(GLPc 组,n=54);组 III,腹腔注射 GLPc 的 IMCD(n=8)。
在组 I 中,PfS 为:ngp-61.8±22.1 和 lgp-8.8±12.4,p<0.01,尿液渗透压为:lgp-735±64 mOsm/Kg 和 ngp-1,632±120 mOsm/Kg。lgp 的 BUN 高于 ngp(176±36 mg%比 56±9 mg%)。在组 II 中,Pf 在 GLPc(250 µg/ml)直接施加于微灌注正常豚鼠 IMCD 浴液的情况下进行测量。GLPc 阻断了 Vp(200 pg/ml,n=5)的作用,不阻断 cAMP(10(-4)M)和 Forskolin(Fors-10(-9)M)的作用,但部分阻断霍乱毒素(ChT-10(-9)M)的作用。来自双曲钩端螺旋体血清型 patoc 的 GLP(GLPp,非致病性,250 µg)不改变 Vp 作用。在组 III 中,腹腔内注射 GLPc(250 µg)使 IMCD 水通道蛋白 2 的表达减少约 20%。
GLP 引起的 IMCD Pf 下降至少部分解释了感染豚鼠观察到的尿浓缩能力丧失。