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苦参碱减轻慢性肾小球肾炎大鼠肾脏损伤及其机制

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目的 探讨苦参碱(Mat)通过调节单核细胞趋化蛋白-1(MCP-1)/趋化因子C-C-基元受体2(CCR2)信号通路对慢性肾小球肾炎大鼠肾脏损伤的影响。方法 采用皮下注射牛血清清蛋白的方式建立慢性肾小球肾炎大鼠模型,将50只大鼠随机均分为对照组、模型组、苦参碱低、高剂量组(Mat-L组、Mat-H组)、苦参碱+CCR2组(Mat+CCR2组),每组10只,连续灌胃4周。检测24 h尿蛋白排泄量;剥离脾脏与胸腺,计算脏器指数;检测各组大鼠血清尿素氮(BUN)、肌酐(Scr)含量;HE染色法观察各组大鼠肾组织病理变化;ELISA法检测肾组织中肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-6水平;流式细胞术检测各组大鼠外周血T淋巴细胞亚群比例;Western blotting法测定MCP-1、CCR2蛋白表达。结果 模型组大鼠脾脏指数和胸腺指数显著降低,肾组织有大量炎症细胞浸润,24 h尿蛋白、BUN、Scr、TNF-α、IL-6以及MCP-1、CCR2表达显著升高,外周血T细胞亚群CD3+、CD4+以及CD4+/CD8+比值显著降低,CD8+比例显著升高(P<0。05);Mat-L组、Mat-H组大鼠脾脏指数和胸腺指数升高,肾组织炎症浸润现象改善,24 h尿蛋白、BUN、Scr、TNF-α、IL-6以及MCP-1、CCR2表达显著降低,外周血T细胞亚群CD3+、CD4+以及CD4+/CD8+比值显著升高,CD8+比例显著降低(P<0。05);CCR2减弱了 Mat-H对慢性肾小球肾炎大鼠免疫功能的影响。结论 苦参碱可以缓解慢性肾小球肾炎大鼠的炎症损伤,提高大鼠外周免疫功能,其作用机制可能与抑制MCP-1/CCR2信号通路有关。
Matrine alleviating kidney injury in rats with chronic glomerulonephritis and its mechanism
Objective To investigate the impact of matrine(Mat)on renal injury in rats with chronic glomerulonephritis by regulating the monocyte chemotactic protein-1(MCP-1)/chemokine C-C-motif receptor 2(CCR2)signaling pathway.Methods The rat model of chronic glomerulonephritis was established by subcutaneous injection of bovine serum albumin,the rats were randomly grouped into control group,model group,low and high dose matrine groups(Mat-L group,Mat-H group),and matrine+CCR2 group(Mat+CCR2 group),with 10 rats in each group.After continuous gavage for 4 weeks,24-hour urine protein excretion was measured;spleen and thymus were removed and organ index was calculated;The levels of blood urea nitrogen(BUN)and serum creatinine(Scr)in the serum of each group were detected,the pathological changes of renal tissue were observed by HE staining;the levels of tumor necrosis factor α(TNF-α)and interleukin(IL)-6 in renal tissue were detected by ELISA;Flow cytometry was applied to detect the proportion of T lymphocyte subsets in peripheral blood of rats in each group;Western blotting was applied to determine the expression of MCP-1 and CCR2 proteins.Results The spleen index and thymus index of rats in the model group rats were obviously reduced,and there were a large number of inflammatory cell infiltration in the renal tissue,the 24-hour urine protein,BUN,Scr,TNF-α,IL-6,and the expression of MCP-1 and CCR2 obviously increased,the peripheral blood T cell subpopulations CD3+,CD4+,and CD4+/CD8+ratio obviously reduced,the proportion of CD8+obviously increased(P<0.05);The spleen index and thymus index of rats in the Mat-L and Mat-H groups of rats increased,and the phenomenon of renal tissue inflammation and infiltration improved,the 24-hour urine protein,BUN,Scr,TNF-α,IL-6,and the expression of MCP-1 and CCR2 obviously decreased,the peripheral blood T cell subpopulations CD3+,CD4+,and CD4+/CD8+ratio obviously increased,the proportion of CD8+obviously decreased(P<0.05);CCR2 attenuated the effect of Mat-H on immune function in rats with chronic glomerulonephritis.Conclusion Matrine can alleviate the inflammatory damage and improve the peripheral immune function of rats with chronic glomerulonephritis,and its mechanism may be related to the inhibition of MCP-1/CCR2 signaling pathway.

MatrineBovine serum albuminMonocyte chemotactic protein-1/chemokine receptor 2 signaling pathwayChronic glomerulonephritisFlow cytometryWestern blottingRat

魏环兵、李昂、高莹、历伟、代云峰

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齐齐哈尔医学院附属第二医院检验科,黑龙江 齐齐哈尔 161006

苦参碱 牛血清清蛋白 单核细胞趋化蛋白-1/趋化因子受体2信号通路 慢性肾小球肾炎 流式细胞术 免疫印迹法 大鼠

2024

解剖学报
中国解剖学会

解剖学报

CSTPCD
影响因子:0.462
ISSN:0529-1356
年,卷(期):2024.55(6)