目的 观察参地颗粒对阳离子化牛血清白蛋白（cationic bovine serum albumin，CBSA）诱导的肾炎大鼠肾脏内质网应激及足细胞自噬标志蛋白的干预作用。方法 将40只雄性SD大鼠随机分为正常组、模型组、参地颗粒组、缬沙坦组，每组10只。参地颗粒组每日给予参地颗粒水溶液4.0 g/kg灌胃，缬沙坦组每日给予缬沙坦胶囊混悬液20 mg/kg灌胃，正常组、模型组予生理盐水灌胃，比色法检测各组大鼠24 h尿蛋白（24hour urine protein,24hUPro）含量，Western blot法检测肾脏中葡萄糖调节蛋白78（glucose regulated protein 78,GRP78)、人内质网应激相关蛋白（human endoplasmic reticulum stressrelated protein,CHOP）、Nephrin、Podocalyxin、Beclin1、微管相关蛋白轻链3（microtubule associated protein light chain 3,LC3)Ⅱ/LC3Ⅰ蛋白表达水平。透射电子显微镜观察各组大鼠肾组织肾小球损伤情况。结果 与正常组比较，模型组大鼠24hUPro显著升高（P＜0.05），肾组织中GRP78、CHOP、Beclin1蛋白相对表达水平和LC3Ⅱ/LC3Ⅰ比值均显著增加，Nephrin、Podocalyxin蛋白相对表达水平显著减少（P＜0.05）。与模型组比较，参地颗粒组和缬沙坦组大鼠24hUpro含量显著降低（P＜0.05），肾组织中GRP78、CHOP表达水平均显著降低，而Nephrin和Podocalyxin表达水平则显著升高（P＜0.05）；参地颗粒组Beclin1表达水平、LC3Ⅱ/LC3Ⅰ比值与模型组比较也进一步增加（P＜0.05），而缬沙坦组增加不够显著（P＞0.05）；且参地颗粒组GRP78、CHOP、Podocalyxin、Beclin1表达水平和LC3Ⅱ/LC3Ⅰ比值变化均显著优于缬沙坦组（P＜0.05）。结论 CBSA肾炎大鼠出现内质网应激反应增强及足细胞损伤，而足细胞自噬活性也增强；参地颗粒能够通过增强自噬活性，减轻内质网应激所介导的足细胞损伤，进而降低CBSA肾炎大鼠尿蛋白水平。
Objective To investigate the intervention effect of the traditional Chinese medicine Shendi Granule on renal endoplasmic reticulum stress and podocyte autophagy in rats with nephritis induced by cationic bovine serum albumin (C-BSA). Methods A total of 40 male Sprague-Dawley rats were randomly divided into normal group, model group, Shendi Granule group, and valsartan group, with 10 rats in each group. The rats in the Shendi Granule group were given Shendi Granule solution 4.0 g/kg by gavage every day, those in the valsartan group were given the suspension of valsartan capsules 20 mg/kg by gavage every day, and those in the normal group and the model group were given normal saline by gavage. Colorimetry was used to measure 24-hour urinary protein (24hUPro), and Western blot was used to measure the protein expression levels of glucose-regulated protein 78 (GRP78), CHOP, nephrin, podocalyxin, and Beclin 1 and microtubule-associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ)/microtubule-associated protein 1 light chain 3-Ⅰ (LC3-Ⅰ) ratio in the kidney. A transmission electron microscope was used to observe glomerular injury in renal tissue. Results Compared with the normal group, the model group had a significant increase in 24hUPro (P<0.05), significant increases in the relative protein expression levels of GRP78, CHOP, and Beclin 1 and LC3-Ⅱ/LC3-Ⅰ ratio in renal tissue (P<0.05), and significant reductions in the relative protein expression levels of nephrin and podocalyxin (P<0.05). Compared with the model group, the Shendi Granule group and the valsartan group had a significant reduction in 24hUPro (P<0.05), significant reductions in the expression levels of GRP78 and CHOP (P<0.05), and significant increases in the protein expression levels of nephrin and podocalyxin (P<0.05); compared with the model group, the Shendi Granule group had significant increases in the expression level of Beclin 1 and LC3-Ⅱ/LC3-Ⅰ ratio (P<0.05), and the valsartan group showed slight increases (P>0.05); compared with the valsartan group, the Shendi Granule group had significantly greater changes in the expression levels of GRP78, CHOP, podocalyxin, and Beclin 1 and LC3-Ⅱ/LC3-Ⅰ ratio (P<0.05). Conclusion Enhanced endoplasmic reticulum stress and podocyte injury are observed in rats with C-BSA nephritis, and there is also a significant increase in podocyte autophagy activity. The traditional Chinese medicine Shendi Granule can enhance autophagy activity and alleviate podocyte injury mediated by endoplasmic reticulum stress, thereby reducing the level of urinary protein in rats with C-BSA nephritis.
魏 玲,金 华,王亿平.参地颗粒对膜性肾病大鼠肾脏内质网应激及足细胞自噬的干预作用[J].安徽中医药大学学报,2022,41(5):90-94复制