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中国医药导报  2023, Vol. 20 Issue (35): 24-30    DOI: 10.20047/j.issn1673-7210.2023.35.05
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参芪蛭龙汤对膜性肾病模型小鼠肾组织中FcγRⅠ、FcγRⅡB、FcγRⅢA表达的影响
薛丕良1  王向伟1  李星宇1  王文文1  王 顺1  刘 星2
1.黑龙江省中医药科学院针灸科,黑龙江哈尔滨 150036;
2.黑龙江省中医药科学院老年病科,黑龙江哈尔滨 150036
Effect of Shenqi Zhilong Decoction on expression of FcγRⅠ, FcγRⅡB, and FcγRⅢA in renal tissue of membranous nephropathy model mice
XUE Piliang1 WANG Xiangwei1 LI Xingyu1 WANG Wenwen1 WANG Shun1 LIU Xing2
1.Department of Acupuncture and Moxibustion, Heilongjiang College of Traditional Chinese Medicine, Heilongjiang Province, Harbin 150036, China;
2.Department of Geriatric, Heilongjiang College of Traditional Chinese Medicine, Heilongjiang Province, Harbin 150036, China
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摘要 目的 探讨参芪蛭龙汤对膜性肾病(MN)模型小鼠肾组织中FcγRⅠ、FcγRⅡB、FcγRⅢA表达的影响。方法 60只SPF级ICR小鼠,6周龄,体重(23±4)g,雌雄各半,按照随机数字表法分为空白组(10只)和造模组(50只)。造模组小鼠注射阳离子化牛血清白蛋白,复制MN小鼠模型。取其中造模成功的40只小鼠按照随机数字表法分为模型组,参芪蛭龙汤高、低剂量组(24、12 g/kg)和雷公藤多苷片组(14 mg/kg),每组10只。给药4周后,处死小鼠。HE、Masson染色观察肾脏组织病理变化;免疫荧光法观察肾组织中FcγRⅠ、FcγRⅡB、FcγRⅢA蛋白荧光强度;Western blot检测肾脏组织内FcγRⅠ、FcγRⅡB、FcγRⅢA蛋白表达;RT-PCR检测肾脏组织中FcγRⅠ、FcγRⅡB、FcγRⅢA mRNA表达。 结果 HE染色显示空白组小鼠上皮细胞、基底膜、肾小管及肾间质形态结构均未见明显异常。模型组小鼠肾小球体积增大,肾小球毛细血管袢扩张,血细胞溢出,肾小球系膜增厚,肾小囊腔增大,局部肾小囊腔壁上皮严重破损。与模型组比较,参芪蛭龙汤高剂量组小鼠可见肾小球体积减小,基底膜增厚减轻,肾小囊壁上皮未破损或局部破损。Masson染色显示空白组小鼠肾小球基底膜正常。模型组小鼠上皮下大量嗜复红蛋白沉积,可见基底膜基质反应,肾小管空泡样脂肪变性,肾间质淋巴大量增生。与模型组比较,各给药组小鼠病变减轻,以参芪蛭龙汤高剂量组改善更加明显。与空白组比较,模型组FcγRⅠ、FcγRⅢA蛋白荧光增强,FcγRⅡB蛋白荧光减弱(P<0.05);与模型组比较,参芪蛭龙汤高剂量组FcγRⅠ、FcγRⅢA蛋白荧光减弱(P<0.05),参芪蛭龙汤高、低剂量组FcγRⅡB蛋白荧光增强(P<0.05);与参芪蛭龙汤低剂量组比较,参芪蛭龙汤高剂量组FcγRⅠ、FcγRⅢA蛋白荧光减弱,FcγRⅡB蛋白荧光增强(P<0.05)。与空白组比较,模型组FcγRⅠ、FcγRⅢA蛋白表达升高,FcγRⅡB蛋白表达降低(P<0.05);与模型组比较,参芪蛭龙汤高剂量组FcγRⅠ、FcγRⅢA蛋白表达降低,参芪蛭龙汤高、低剂量组FcγRⅡB蛋白表达升高(P<0.05);与参芪蛭龙汤低剂量组比较,参芪蛭龙汤高剂量组FcγRⅡB蛋白表达升高,FcγRⅢA蛋白表达降低(P<0.05)。与空白组比较,模型组FcγRⅠ、FcγRⅢA mRNA表达升高,FcγRⅡB mRNA表达降低(P<0.05);与模型组比较,参芪蛭龙汤高、低剂量组FcγRⅠ、FcγRⅢAmRNA表达降低,FcγRⅡB mRNA表达升高(P<0.05);与参芪蛭龙汤低剂量组比较,参芪蛭龙汤高剂量组FcγRⅠ、FcγRⅢA mRNA表达降低,FcγRⅡB mRNA表达升高(P<0.05)。 结论 参芪蛭龙汤对MN小鼠的治疗作用与改善肾组织病理改变,减少FcγRⅠ、FcγRⅢA蛋白的表达,增加FcγRⅡB蛋白表达有关,其中高浓度的参芪蛭龙汤效果更好。
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薛丕良1  王向伟1  李星宇1  王文文1  王 顺1  刘 星2
关键词 膜性肾病参芪蛭龙汤阳离子牛血清白蛋白FcγRⅠFcγRⅡBFcγRⅢA    
Abstract:Objective To explore the effect of Shenqi Zhilong Decoction on expression of FcγRⅠ, FcγRⅡB, and FcγRⅢA in renal tissues of membranous nephropathy (MN) model mice. Methods Sixty SPF grade ICR mice, six-week old and weig- hting (23±4) g, half male and half female, were divided into blank group (10 mice) and model group (50 mice). The model group mice were injected with cationic-bovine serum albumin to replicate MN mouse model. The 40 mice successfully modeled were divided into model group, Shenqilong Decoction high-, low-dose group (24, 12 g/kg) and Tripterygium Glycoside Tablets group (14 mg/kg) according to the random number table method, with 10 mice in each group. After four weeks of administration, the mice were killed. HE and Masson staining were used to observe the pathological changes of renal tissues; immunofluorescence method was used to observe the protein fluorescence intensity of FcγRⅠ, FcγRⅡB, and FcγRⅢA in renal tissues; Western blot was used to detect the protein expression of FcγRⅠ, FcγRⅡB, and FcγRⅢA in renal tissues; the mRNA expression of FcγRⅠ, FcγRⅡB, and FcγRⅢA in renal tissues was detected by RT-PCR. Results HE staining showed no significant abnormalities in the morphology and structure of the epithelial cells, basement membrane, renal tubules, and renal interstitium in blank group. In model group, the glomerular volume increased, the glomerular capillary loop dilated, blood cell overflow, the mesangial thickened, the cystic cavity enlarged, and the skin on the wall of the cystic cavity was severely damaged. Compared with model group, the glomerular volume decreased, the basement membrane thickened and the skin on the wall of the renal capsule was not damaged or partially damaged in Shenqi Zhilong Decoction high-dose group. Masson staining showed that the glomerular basement membrane was normal in the blank group. In model group, there was a large amount of subepithelial photophilin deposition, basal membrane matrix reaction, vacuol-like steatosis of renal tubules, and a large amount of renal interstitial lymphatic hyperplasia. Compared with model group, the lesion of each administration group was reduced, and the improvement was more obvious in Shenqi Zhilong Decoction high-dose group. Compared with blank group, the protein fluorescence of FcγRⅠ and FcγRⅢA in model group was enhanced, while the protein fluorescence of FcγRⅡB was weakened (P<0.05); compared with model group, the protein fluorescence of FcγRⅠ and FcγRⅢA in Shenqi Zhilong Decoction high-dose group was decreased (P<0.05), while the protein fluorescence of FcγRⅡB in Shenqi Zhilong Decoction high- and low-dose group was increased (P<0.05); compared with Shenqi Zhilong Decoction low-dose group, the protein fluorescence of FcγRⅠ and FcγRⅢA in Shenqi Zhilong Decoction high-dose group was decreased, and the protein fluorescence of FcγRⅡB was increased (P<0.05). Compared with blank group, the protein expressions of FcγRⅠ and FcγRⅢA were increased in model group, while the protein expression of FcγRⅡB was decreased (P<0.05); compared with model group, the protein expressions of FcγRⅠ and FcγRⅢA in Shenqi Zhilong Decoction high-dose group were decreased, while the protein expressions of FcγRⅡB in Shenqi Zhilong Decoction high- and low-dose groups were increased (P<0.05); compared with Shenqi Zhilong Decoction low-dose group, the protein expression of FcγRⅡB in Shenqi Zhilong Decoction high-dose group was increased, and the protein expression of FcγRⅢA was decreased (P<0.05). Compared with blank group, the mRNA expressions of FcγRⅠ and FcγRⅢA in model group were increased, while the mRNA expressions of FcγRⅡB was decreased (P<0.05); compared with model group, the mRNA expressions of FcγRⅠ and FcγRⅢA in Shenqi Zhilong Decoction high- and low-dose groups were decreased, and the mRNA expression of FcγRⅡB was increased (P<0.05). compared with Shenqi Zhilong Decoction low-dose group, the mRNA expressions of FcγRⅠ and FcγRⅢA in Shenqi Zhilong Decoction high-dose group were decreased, while the mRNA expression of FcγRⅡB were increased (P<0.05). Conclusion The therapeutic effect of Shenqi Zhilong Decoction on MN mice is related to improving the pathological changes of renal tissues, reducing the expressions of FcγRⅠ and FcγRⅢA, and increasing the protein expression of FcγRⅡB, and the effect of Shenqi Zhilong Decoction with high concentration is better.
Key wordsMembranous nephropathy    Shenqi Zhilong Decoction    Cationic-bovine serum albumin    FcγRⅠ    FcγRⅡB    FcγRⅢA
    
基金资助:黑龙江省自然科学基金项目(LH2020H104)。
通讯作者: 王顺(1965.8-),男,医学博士,主任医师,博士生导师,主要从事针灸治疗内科疾病的研究。 刘星(1985.10-),女,硕士,副主任护师,黑龙江省中医药科学院老年病科护士长,主要从事老年性疾病的中西医结合护理工作。   
作者简介: 薛丕良(1983.9-),男,博士,副主任医师,硕士生导师,主要从事内科疾病的中西医诊疗工作。
引用本文:   
薛丕良1  王向伟1  李星宇1  王文文1  王 顺1  刘 星2. 参芪蛭龙汤对膜性肾病模型小鼠肾组织中FcγRⅠ、FcγRⅡB、FcγRⅢA表达的影响[J]. 中国医药导报, 2023, 20(35): 24-30.
XUE Piliang1 WANG Xiangwei1 LI Xingyu1 WANG Wenwen1 WANG Shun1 LIU Xing2. Effect of Shenqi Zhilong Decoction on expression of FcγRⅠ, FcγRⅡB, and FcγRⅢA in renal tissue of membranous nephropathy model mice. 中国医药导报, 2023, 20(35): 24-30.
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