参地颗粒对系膜增生性肾小球肾炎大鼠单个核细胞凋亡及炎症紊乱的干预作用
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安徽省卫计委中医药科研项目(2016ZY13);安徽中医药领军人才项目(中医药发展秘〔2018〕23号);安徽省高校自然科学研究重点项目(KJ2019A0441)


Interventional Effect of Shendi Granule on Apoptosis and Inflammatory Disorder of Mononuclear Cells in Rats with Mesangial Proliferative Glomerulonephritis
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    目的 观察参地颗粒作用于系膜增生性肾小球肾炎(mesangial proliferative glomerulonephritis,MsPGN)大鼠后,外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)的凋亡,B淋巴细胞瘤- 2(B- cell lymphoma- 2,Bcl- 2)蛋白、Fas蛋白的表达,单核细胞趋化蛋白- 1(monocyte chemotactic protein 1,MCP- 1)、转化生长因子- β1(transforming growth factor- β1,TGF- β1)水平的变化,探讨其作用机制。方法 从60只SD大鼠中随机抽取10只作为正常组,剩余50只均用于MsPGN模型复制,模型制作成功后,再分为模型组(11只)、缬沙坦组(10只)、参地颗粒组(10只)。各组大鼠在模型复制完成后第21天开始给药,缬沙坦组每日予缬沙坦溶液灌胃,参地颗粒组予参地颗粒溶液灌胃,模型组、正常组予生理盐水灌胃,连续干预12周。光学显微镜下观察各组大鼠病理损伤情况,实验室方法检测各组大鼠24 h尿蛋白(24- hour urinary protein,24hUPr)、PBMCs凋亡率、Bcl- 2蛋白、Fas蛋白表达水平及血清MCP- 1、TGF- β1水平。结果 治疗第8、12周参地颗粒组24hUPr水平较缬沙坦组显著降低(P<0.05)。与正常组比较,模型组大鼠PBMCs凋亡率、血清MCP- 1、TGF- β1水平升高(P<0.05),Bcl- 2蛋白表达水平下降,Fas蛋白表达水平上升(P<0.05);与模型组比较,缬沙坦组、参地颗粒组PBMCs凋亡率、MCP- 1、TGF- β1水平均降低(P<0.05),Bcl- 2蛋白表达水平上升,Fas蛋白表达水平下降(P<0.05);参地颗粒组PBMCs凋亡率、血清TGF- β1水平低于缬沙坦组(P<0.05),Bcl- 2蛋白表达水平上升程度大于缬沙坦组(P<0.05)。参地颗粒组MsPGN大鼠肾脏系膜细胞增生被明显抑制。结论 参地颗粒可以减轻肾脏炎症反应,消减尿蛋白,改善MsPGN大鼠的肾脏病理损害,其机制可能是通过调节凋亡调控蛋白Bcl- 2、Fas蛋白表达水平,降低MsPGN大鼠PBMCs凋亡率,下调MCP- 1、TGF- β1的表达水平实现的。

    Abstract:

    Objective To investigate the changes in the apoptosis of peripheral blood mononucleated cells (PBMCs), the protein expression of B- cell lymphoma- 2 (Bcl- 2) and Fas, and the levels of monocyte chemotactic protein 1 (MCP- 1) and transforming growth factor- β1 (TGF- β1) after Shendi Granule is used for the treatment of rats with mesangial proliferative glomerulonephritis (MsPGN), as well as the mechanism of action of Shendi Granule. Methods A total of 60 Sprague- Dawley rats were selected, among which 10 rats were randomly selected as normal group and the remaining 50 rats were used to establish a model of MsPGN, and after the model was successfully established, they were divided into model group with 11 rats, valsartan group with 10 rats, and Shendi Granule group with 10 rats. The drugs were administered since day 21 after modeling; the rats in the valsartan group were given valsartan solution by gavage every day, those in the Shendi Granule group were given Shendi Granule solution by gavage, and those in the model group and the normal group were given normal saline by gavage, for 12 consecutive weeks. Pathological damage was observed under a light microscope, and laboratory methods were used to measure 24- hour urinary protein (24hUPr), apoptosis rate of PBMCs, protein expression of Bcl- 2 and Fas, and serum levels of MCP- 1 and TGF- β1. Results Compared with the valsartan group, the Shendi Granule group had a significantly greater reduction in 24hUPr at weeks 8 and 12 of treatment (P<0.05). Compared with the normal group, the model group had significant increases in the apoptosis rate of PBMCs and the serum levels of MCP- 1 and TGF- β1 (P<0.05), as well as significantly downregulated protein expression of Bcl- 2 and significantly upregulated protein expression of Fas (P<0.05). Compared with the model group, both the valsartan group and the Shendi Granule group had significant reductions in the apoptosis rate of PBMCs and the serum levels of MCP- 1 and TGF- β1 (P<0.05), as well as significantly upregulated protein expression of Bcl- 2 and significantly downregulated protein expression of Fas (P<0.05). Compared with the valsartan group, the Shendi Granule group had significantly lower apoptosis rate of PBMCs and serum level of TGF- β1 (P<0.05) and a significantly greater increase in the protein expression of Bcl- 2 (P<0.05). The proliferation of renal mesangial cells was effectively inhibited in the MsPGN rats in the Shendi Granule group. Conclusion Shendi Granule can alleviate renal inflammatory response, reduce urinary protein, and improve the pathological damage of the kidney in rats with MsPGN, possibly by regulating the protein expression of the apoptosis- regulating proteins Bcl- 2 and Fas, reducing the apoptosis rate of PBMCs, and downregulating the expression levels of MCP- 1 and TGF- β1.

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茅燕萍,王恩静,金 华,魏 玲,张 磊,王亿平.参地颗粒对系膜增生性肾小球肾炎大鼠单个核细胞凋亡及炎症紊乱的干预作用[J].安徽中医药大学学报,2021,40(3):70-74

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  • 在线发布日期: 2021-06-16