临床肾脏病杂志2025，Vol.25Issue(1) ：50-58.DOI:10.3969/j.issn.1671-2390.2025.01.008

神经轴突导向分子3对糖尿病肾脏疾病大鼠肾脏纤维化的影响

Effect of Slit3 on kidney fibrosis in diabetic kidney disease rats

黄明芳 董杨 哈力努尔·沙德克江 甘馨雨 史文丽 陆晨
临床肾脏病杂志2025，Vol.25Issue(1) ：50-58.DOI:10.3969/j.issn.1671-2390.2025.01.008
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神经轴突导向分子3对糖尿病肾脏疾病大鼠肾脏纤维化的影响

Effect of Slit3 on kidney fibrosis in diabetic kidney disease rats

黄明芳 1董杨 1哈力努尔·沙德克江 1甘馨雨 1史文丽 2陆晨1
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作者信息

  • 1. 新疆医科大学第一附属医院肾脏疾病中心,乌鲁木齐 830054
  • 2. 新疆维吾尔自治区人民医院肾脏病科,乌鲁木齐 830001
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摘要

目的 探讨神经轴突导向分子3(slit guidance ligand 3,Slit3)对糖尿病肾脏疾病(dia-betic kidney disease,DKD)大鼠肾脏纤维化的影响.方法 采用高糖高脂饮食联合多次小剂量注射链脲佐菌素建造DKD大鼠模型,将40只SD大鼠采用随机数字表法分为空白对照(control)组、糖尿病(diabetes mellitus,DM)组、DKD4周组、DKD8周组,每组各10只.造模成功后,control组剩8只,其余各组剩6只.检测24 h尿蛋白定量;腹主动脉取血,分析总胆固醇、三酰甘油、尿素氮(blood urea nitro-gen,BUN)、血肌酐(serum creatinine,Scr)、白蛋白;HE染色观察肾脏组织病理学改变;Masson染色观察肾脏组织胶原纤维的沉积;采用实时荧光定量聚合酶链反应法检测肾脏组织中Slit3、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、抗转化生长因子β1(transforming growth factor-β1,TGF-β1)mRNA的表达水平;采用蛋白质印迹法检测肾脏组织中Slit3、α-SMA、TGF-β1蛋白表达水平;免疫组织化学染色法检测肾脏组织中的Slit3表达和定位情况.结果 与control组相比,DKD4周组、DKD8周组的24 h尿蛋白定量[(37.18±5.55)mg、(47.38±18.19)mg比(10.66±4.87)mg]、BUN[(19.18±7.97)mmol/L、(21.01±6.74)mmol/L比(6.86±1.15)mmol/L]、Scr[(58.02±12.49)μmol/L、(61.18±20.76)μmol/L比(28.34±3.35)μmol/L]均显著升高(P＜0.05),TC[(3.75±3.21)mmol/L、(2.44±2.08)mmol/L比(1.37±0.32)mmol/L]升高而TG[(0.46±0.64)mmol/L、(0.32±0.50)mmol/L比(0.93±0.46)mmol/L]降低(P＜0.05).与control组相比,DKD4周组、DKD8周组大鼠肾脏组织中Slit3[(0.100±0.002)、(0.120±0.003)比(0.050±0.006)]、α-SMA[(0.370±0.011)、(0.590±0.003)比(0.070±0.006)]、TGF-β1[(0.810±0.024)、(0.940±0.01)比(0.520±0.028)]蛋白表达水平和Slit3[(2.520±0.164)、(3.016±0.226)比(1.020±0.034)]、α-SMA[(3.010±0.160)、(3.560±0.218)比(1.000±0.100)]、TGF-β1[(3.030±0.185)、(3.550±0.194)比(1.010±0.087)]mRNA表达水平均升高(P＜0.05);与DM组相比,DKD4周组、DKD8周组肾脏组织中Slit3[(0.100±0.002)、(0.120±0.003)比(0.070±0.004)]、α-SMA[(0.370±0.011)、(0.590±0.003)比(0.220±0.014)]、TGF-β1[(0.810±0.024)、(0.940±0.010)比(0.700±0.028)]蛋白表达水平明显升高(P＜0.05),尤以DKD8周组升高更明显(P＜0.01).而与DKD4周组相比,DKD8周组肾脏组织中各mRNA表达水平差异均无统计学意义(P＞0.05).结论 Slit3显著加剧DKD大鼠肾脏纤维化.

Abstract

Objective To explore the effect of slit guidance ligand 3(Slit3)on rats with diabetic kidney disease(DKD)-induced fibrosis.Methods A DKD rat model was established by multiple low-dose injections of streptozotocin combined with high-sugar high-fat diet.Forty Sprague-Dawley(SD)rats were randomly divided into the blank control group(control),diabetes mellitus(DM)group,4-week DKD group and 8-week DKD group,with 10 rats in each group by random number table method.After successful modeling,there were 8 rats in the control group,and 6 in each of the remaining groups.The 24 h urinary protein content was detected.Total cholesterol(TC),triglyceride(TG),blood urea nitrogen(BUN),serum creatinine(Scr),and albumin(Alb)were analyzed after abdominal aorta blood collection.Hematoxylin and eosin(H&E)staining was used to observe the histopathological changes of the kidney.Kidney fibrosis degree was examined by Masson's trichrome staining.The reverse-transcription quantitative real-time polymerase chain reaction(RT-qPCR)was performed to detect mRNA levels of Slit3,α-smooth muscle actin(α-SMA),and transforming growth factor-β1(TGF-β1)in kidney tissues,and their protein levels were detected by Western blot.The expression and localization of Slit3 in kidney tissues were detected by immunohistochemical staining.Results Compared with the control group,24-hour urinary protein content[(37.18±5.55)mg、(47.38±18.19)mg vs(10.66±4.87)mg],BUN[(19.18±7.97)mmol/L、(21.01±6.74)mmol/L vs(6.86±1.15)mmol/L]and Scr[(58.02±12.49)μmol/L、(61.18±20.76)μmol/L vs(28.34±3.35)μmol/L]of 4-week and 8-week DKD groups were significantly higher(P＜0.05).In comparison to the control group,TC[(3.75±3.21)mmol/L、(2.44±2.08)mmol/L vs(1.37±0.32)mmol/L]was significantly higher,while TG[(0.46±0.64)mmol/L、(0.32±0.50)mmol/L vs(0.93±0.46)mmol/L]was significantly lower in the 4-week and 8-week DKD groups(P＜0.05).Compared with the control group,the protein levels of Slit3[(0.100±0.002)、(0.120±0.003)vs(0.050±0.006)],α-SMA[(0.370±0.011)、(0.590±0.003)vs(0.070±0.006)]and TGF-β1[(0.810±0.024)、(0.940±0.01)vs(0.520±0.028)]in rat kidney tissue were significantly higher in the 4-week and 8-week DKD groups(P＜0.05);so as their mRNA levels[(2.520±0.164)、(3.016±0.226)vs(1.020±0.034);(3.010±0.160)、(3.560±0.218)vs(1.000±0.100);(3.030±0.185)、(3.550±0.194)vs(1.010±0.087)].Compared with the DM group,the protein levels of Slit3[(0.100±0.002)、(0.120±0.003)vs(0.070±0.004)],α-SMA[(0.370±0.011)、(0.590±0.003)vs(0.220±0.014)]and TGF-β1[(0.810±0.024)、(0.940±0.010)vs(0.700±0.028)]in rat kidney tissue were significantly higher in the 4-week and 8-week DKD groups(P＜0.05),especially in the 8-week DKD group(P＜0.01).No significant differences in the mRNA levels of Slit3,α-SMA and TGF-β1 were examined in the rat kidney tissue between 4-week and 8-week DKD groups(P＞0.05).Conclusion Slit3 can signifi-cantly aggravate kidney fibrosis in DKD rats.

关键词

神经轴突导向分子3/糖尿病肾脏疾病/肾脏纤维化

Key words

Slit guidance ligand 3/Diabetic kidney disease/Kidney fibrosis

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出版年

2025
临床肾脏病杂志
中华医学会武汉分会，湖北省微循环学会

临床肾脏病杂志

影响因子：0.573
ISSN：1671-2390
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