Changes of kidney in rat models of type 2 diabetes mellitus after administering Lizhong Decoction

doi:10.3969/j.issn.2095-4344.1385

Abstract

Abstract:

BACKGROUND: Lizhong Decoction is a common prescription for treating diabetic nephropathy. However, the mechanism of Lizhong Decoction on the prevention and treatment of diabetic nephropathy is still unclear.
OBJECTIVE: To confirm that Lizhong Decoction has renal protective effect on type 2 diabetic rats and the underlying mechanism by exploring the changes of renal function, renal tissue morphology and expression of renal cortex vascular endothelial growth factor, transforming growth factor-β1, and type I collagen proteins.
METHODS: Sixty 7-8-week-old male Sprague-Dawley rats, SPF level, were provided by Laboratory Animal Center of Guangzhou University of Chinese Medicine (license No: SCXK2013-0034), and the study was approved by the Experimental Animal Ethics Committee of the First Affiliated Hospital of Guangzhou University of Chinese Medicine. The rats were divided into control and modeling group. Rats in the modeling group were given high-sugar and high-fat diet to induce diet-induced obesity. These rats were then injected with low-dose streptozotocin (28 mg/kg), and observed for 4 consecutive weeks, and fasting blood glucose ≥ 11.1 mmol/L was included in type 2 diabetic rats. Afterwards, the diabetic rats were divided into model, chatain and Lizhong groups, and were administered for 12 consecutive weeks. The renal biochemical parameters, renal histomorphological changes and expression of renal cortex vascular endothelial growth factor A, transforming growth factor-β1, and type I collagen proteins were detected.
RESULTS AND CONCLUSION: (1) The kidney weight index in the Lizhong group was significantly lower than that in the model group (P < 0.05). (2) The fasting blood glucose, total cholesterol, triglyceride, serum creatinine, urea nitrogen, insulin, insulin resistance index, glycosylated hemoglobin, and urinary microalbumin in the Lizhong group were significantly lower than those in the model group (P < 0.05). (3) The pathological changes of renal tissue in the Lizhong group were significantly reduced compared with the model group. The staining area of PAS, PASM and MASSON staining positive substances in the Lizhong group were all smaller than those in the model group (P < 0.05). (4) The expression levels of vascular endothelial growth factor A, transforming growth factor-β1, and type I collagen proteins in renal cortex in the Lizhong group were significantly lower than those in the model group (P < 0.05). (5) These results indicate that Lizhong Decoction has renal protective effect on type 2 diabetic rats, and its mechanism may be related to reduction in the glucose and lipid and down-regulated expression of vascular endothelial growth factor A, transforming growth factor-β1, and type I collagen proteins.

Key words: Lizhong Decoction, type 2 diabetes, diabetic nephropathy, vascular endothelial growth factor A, transforming growth factor-β1, type I collagen, the National Natural Science Foundation of China

CLC Number:

Liang Lichang, , Fan Yawen, , Mu Lei, , Xie Tian, , Jiang Xiaobing, Ren Hui, Zhang Tianfeng. Changes of kidney in rat models of type 2 diabetes mellitus after administering Lizhong Decoction[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(27): 4356-4362.

Figures/Tables 3

2.1 实验动物数量分析实验选用大鼠60只，分为空白组10只和造模组50只，剔除不符合食源性肥胖9只及不符合2型糖尿病模型的2只大鼠，其余49只大鼠(分为4组)进入结果分析。 2.2 大鼠一般情况、体质量和肾脏质量指数正常组大鼠毛色洁白有光泽，活动状态良好，反应灵敏，饮食及尿便正常。造模组大鼠于注射链脲佐菌素后出现明显的多饮、多食、多尿，毛色泛黄无光泽、反应稍迟钝等症状。与空白组比较，给药0，4，8，12周时，各组大鼠的体质量均下降(P < 0.05)；与模型组相比，在给药后各时间点理中组和沙坦组大鼠的体质量差异无显著性意义(P > 0.05)。给药12周后，与空白组比较，各组大鼠肾脏质量指数显著升高(P < 0.05)；与模型组比较，理中组和沙坦组大鼠的肾脏质量指数均明显降低(P < 0.05)，但2组间比较无明显差异(P > 0.05)。见表1。 2.3 各项肾脏相关的生化指标给药12周后，与空白组相比，各组的空腹血糖、总胆固醇、三酰甘油、肌酐、尿素氮、胰岛素、胰岛素抵抗指数、糖化血红蛋白和尿微量白蛋白均显著升高(P < 0.05)。与模型组相比，理中组能降低血糖、总胆固醇、三酰甘油、胰岛素、胰岛素抵抗指数和糖化血红蛋白(P < 0.05)，而沙坦组降低作用无显著性意义(P > 0.05)；同时与模型组相比，理中组和沙坦组均可以明显降低肌酐、尿素氮和尿微量白蛋白(P < 0.05), 但2组间比较差异无显著性意义(P > 0.05)。见表2。 2.4 肾脏组织形态学变化 ①4种特殊染色综合分析见图1。空白组肾小球和肾小管形态基本正常，球内毛细血管腔清晰且无扩张，系膜细胞无增生，系膜基质分布正常，有完整的囊壁结构，肾小球和肾小管间隙有少量胶原纤维分布；模型组肾小球出现代偿性扩张或萎缩，系膜细胞明显增生，系膜区增宽，基底膜增厚，部分球囊壁出现粘连，肾小管上皮细胞出现空泡变性，肾小球和肾小管间隙胶原纤维明显增多，出现肾小球硬化；理中组和沙坦组肾脏的病理变化较模型组明显减轻，系膜细胞和系膜基质轻度增生，局部基底膜增厚，肾小管上皮细胞空泡变性减少，肾小球和肾小管间隙胶原纤维明显减少。3种染色阳性率分析：与模型组比较，理中组和沙坦组PAS、PASM、MASSON染色阳性率均显著降低(P < 0.05)；②透射电镜观察见图2。与空白组比较，模型组可见肾小球内系膜细胞增生，系膜区基质增多变宽、基底膜明显增厚，足突排列紊乱，部分足突融合，足突间隙增宽。与模型组比较，理中组和沙坦组系膜细胞、系膜基质、基底膜和足突改变均有所减轻。 2.5 肾皮质血管内皮生长因子A、转化生长因子β1、Ⅰ型胶原蛋白表达与空白组比较，糖尿病各组大鼠血管内皮生长因子A、转化生长因子β1、Ⅰ型胶原蛋白表达升高(P < 0.05)。与模型组比较，理中组和沙坦组血管内皮生长因子A、转化生长因子β1、Ⅰ型胶原蛋白表达明显降低(P < 0.05)，但理中组和沙坦组2组间比较无明显差异(P > 0.05)。见图3。 "

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