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目的:探讨丹芍颗粒Ⅲ号对紫癜性肾炎(HSPN)大鼠的作用机制。方法:将雄性SD大鼠随机分为5组:空白组,模型组,丹芍低、中、高剂量组,各5只,造模后丹芍各剂量组给予丹芍颗粒Ⅲ号治疗,取肾组织后送病理观察;BCA法进行大鼠24h尿蛋白定量;Western blot法检测大鼠肾组织中p-Akt蛋白量;RT-PCR法检测大鼠肾组织Akt mR NA的表达量;透射电子显微镜下观察肾小球基底膜及足细胞。结果:光镜下,模型组肾组织存在明显异常改变;丹芍各剂量组肾组织病变较模型组减轻。24h尿蛋白定量模型组显著高于空白组(P<0.05);丹芍高、中、低剂量组均低于模型组(P<0.05)。与空白组比,模型组p-Akt蛋白灰度值比率显著下降(P<0.01),与模型组比,丹芍高、中剂量组p-Akt蛋白比率均升高(P<0.05)。5组Akt m RNA表达不等,但两两之间差异无统计学意义。空白组肾小球基底膜清晰完整,足突未见融合,而模型组肾小球基底膜偶见增厚,但足突大部分融合,消失。丹芍各剂量组大鼠肾小球基底膜基本光滑均匀,足突融合较模型组明显减轻。结论:丹芍颗粒Ⅲ号诱导Akt蛋白的磷酸化,促进PI3K/Akt信号通路抑制肾小球足细胞融合、凋亡的作用,可能是发挥对大鼠HSPN治疗作用的机制。
Objective: To investigate the mechanism of Dan Shao Zhi No. Ⅲ on purpura nephritis (HSPN) rats. Methods: Male Sprague-Dawley rats were randomly divided into 5 groups: blank group, model group, low, middle and high dosage of Dan Shao, 5 rats each. The pathological changes of renal tissue were observed by transmission electron microscopy. The protein expression of 24h urinary protein was detected by BCA method. The protein level of p-Akt was detected by Western blot. The expression of Akt mRNA was detected by RT-PCR. The glomerular basement membrane and podocytes were observed under a microscope. Results: Under the light microscope, there was a significant abnormal change in the kidney tissue of the model group; the renal tissue lesion in each dose group of Danshen was relieved compared with the model group. 24h urinary protein quantitative model group was significantly higher than the blank group (P <0.05); Dan Shao high, medium and low dose groups were lower than the model group (P <0.05). Compared with the blank group, the gray value of p-Akt protein in the model group decreased significantly (P <0.01). Compared with the model group, the ratio of p-Akt protein in the high and middle dose of Danshao group increased (P <0.05). The expression of Akt m RNA in 5 groups was not equal, but there was no significant difference between every two groups. The glomerulus basement membrane in the blank group was clear and complete, and there was no fusion in the foot process. However, the glomerular basement membrane was occasionally thicker in the model group, but most of the foot process disappeared. Danshao the glomerular basement membrane of each dose group was basically smooth, foot process fusion significantly reduced compared with the model group. Conclusion: Dan Sha particles Ⅲ can induce the phosphorylation of Akt protein and promote PI3K / Akt signaling pathway to inhibit glomerular podocyte fusion and apoptosis, which may be the mechanism of treating rat HSPN.