1.上海中医药大学附属曙光医院肾病科(上海 201203)
张亚亨,男,硕士,住院医师,主要从事中医药治疗肾病临床研究工作
杨雪军,主任医师,研究员,硕士研究生导师; E-mail:yangxuejun@shutcm.edu.cn
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张亚亨,盛广宇,宋婷,等.玉蚕颗粒改善足细胞损伤治疗糖尿病肾病的机制研究[J].上海中医药杂志,2023,57(11):77-84.
ZHANG Yaheng,SHENG Guangyu,SONG Ting,et al.Mechanism of Yucan Granules improving podocyte damage and treating diabetic nephropathy[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(11):77-84.
张亚亨,盛广宇,宋婷,等.玉蚕颗粒改善足细胞损伤治疗糖尿病肾病的机制研究[J].上海中医药杂志,2023,57(11):77-84. DOI: 10.16305/j.1007-1334.2023.2306010.
ZHANG Yaheng,SHENG Guangyu,SONG Ting,et al.Mechanism of Yucan Granules improving podocyte damage and treating diabetic nephropathy[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(11):77-84. DOI: 10.16305/j.1007-1334.2023.2306010.
目的,2,观察玉蚕颗粒对糖尿病肾病(DN)小鼠的治疗作用并探讨其作用机制。,方法,2,将80只SPF级雄性c57小鼠随机分为正常组和造模组。正常组10只小鼠予普通饲料;造模组70只小鼠采用高脂高糖饲料喂养联合链脲佐菌素(STZ)腹腔注射的方法建立DN小鼠模型。将50只造模成功的小鼠再随机分为模型组、氯沙坦钾组(氯沙坦钾10 mg/kg)及中药低、中、高剂量组(玉蚕颗粒1.5、3、6 g/kg),每组10只。给药组予相应药物灌胃处理,正常组及模型组以质量分数为0.9%的氯化钠溶液灌胃,每日1次,持续给药4周。①观察小鼠的一般状态,并比较给药前后的体质量变化;给药结束后检测血清空腹血糖(FBG);②采用苏木精-伊红(HE)染色、丽春红酸性品红-苯胺蓝(Masson)染色、过碘酸-雪夫(PAS)染色观察肾组织病理变化;③采用透射电镜观察肾小球足细胞超微结构变化;④采用免疫组织化学法检测足细胞标志蛋白足细胞裂孔膜蛋白(Nephrin)、肾母细胞瘤基因1蛋白(WT1)阳性表达情况,并采用蛋白质免疫印迹(Western blot)法检测Nephrin、WT1蛋白表达水平。,结果,2,①与正常组比较,模型组小鼠的一般状态较差,体质量下降,FBG升高(,P,<,0.05);与模型组比较,各给药组小鼠状态好转、体质量升高、FBG下降,其中中药中、高剂量组FBG差异有统计学意义(,P,<,0.05)。②HE、Masson及PAS染色显示:与正常组比较,模型组小鼠肾小管管壁变薄、管腔扩张、炎症细胞浸润、肾间质纤维组织增生;与模型组比较,中药各剂量组小鼠肾小管管壁变薄、管腔扩张、纤维组织增生有所减轻、炎症细胞浸润减少。③透射电镜观察显示:与正常组比较,模型组小鼠肾小球足细胞足突相互融合、部分足突消失;与模型组比较,中药各剂量组足突融合、消失情况明显好转,且中药中、高剂量组对足细胞损伤的改善效果较明显。④免疫组织化学染色结果与Western blot结果显示:与正常组比较,模型组肾组织Nephrin、WT1表达显著降低(,P,<,0.05);与模型组比较,中药各剂量组小鼠肾组织Nephrin、WT1表达显著升高(,P,<,0.05)。,结论,2,玉蚕颗粒可通过降低DN小鼠血糖水平、减轻肾脏组织炎症细胞浸润、改善足细胞损伤,从而治疗糖尿病肾病。
Objective,2,To observe the therapeutic effect of Yucan Granules (YCG) on diabetic nephropathy (DN) mice and explore its mechanism.,Methods,2,Eighty SPF male C57 mice were randomly divided into normal group and model group. Ten mice in the normal group were fed with normal diet, and 70 mice in the model group were fed with high-fat and high-sugar diet combined with intraperitoneal injection of streptozotocin (STZ) to establish the model of DN mice. Among them, 50 mice were successfully molded and randomly divided into model group, losartan potassium group (losartan potassium 10 mg/kg) and YCG low, medium and high dose groups (YCG 1.5,3,6 g/kg) with 10 mice in each group. The treatment groups were given corresponding drug by gavage, and the normal group and model group were gavaged with 0.9% sodium chloride solution by mass fraction, once a day. After continuous administration for 4 weeks: ①The general state of mice was observed, and the changes of body mass of mice before and after administration were compared. Fasting blood glucose (FBG) was measured after administration. ②The pathological changes of renal tissue were observed by hematoxylin-eosin (HE) staining, fuchsin-aniline blue (Masson) staining and PAS staining. ③The ultrastructural changes of glomerular podocytes were observed by transmission electron microscope. ④The expressions of nephrin, a podocyte marker protein and nephroblastoma gene 1 (WT1) protein were detected by immunohistochemistry and Western blot.,Results,2,①Compared with the normal group, the general state of the model group mice was poor, with a decrease in body mass and an increase in FBG (,P,<,0.05); Compared with the model group, the state of mice in each treatment group improved, their body mass increased, and their FBG decreased. There was a statistically significant difference in FBG between the medium and high dose groups of YCG (,P,<,0.05). ②HE, Masson and PAS staining showed that compared with the normal group, the renal tubule wall of the model group became thinner, the lumen dilated, the inflammatory cells infiltrated, the renal interstitial fibrous tissue proliferated. Compared with the model group, the wall of renal tubule became thinner, the lumen dilated, the proliferation of fibrous tissue decreased, the infiltration of inflammatory cells decreased in the YCG groups. ③Transmission electron microscopy observations showed that: compared with the normal group, the foot processes of glomerular podocytes in the model group merged with each other and some foot processes disappeared; Compared with the model group, the fusion and disappearance of podocytes in the YCG groups were obviously improved, and the improvement effect of podocyte damage in the YCG medium and high dose groups was obvious. ④The results of immunohistochemical staining and Western blot showed that compared with the normal group, the expression of Nephrin and WT1 proteins in the renal tissue was significantly decreased in the model group (,P,<,0.05); Compared with the model group, the expression of Nephrin and WT1 proteins in the renal tissue was significantly increased in the YCG groups (,P,<,0.05).,Conclusion,2,YCG can treat DN by reducing the level of blood glucose in DN mice and infiltrations of the inflammatory cells in renal tissue, and protecting damaged podocyte.
糖尿病肾病玉蚕颗粒足细胞损伤作用机制小鼠模型中药研究
diabetic nephropathyYucan Granulepodocyte damagemechanism of actionmice modeltraditional Chinese herbal medicine research
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