滋阴清热活血方联合激素对系统性红斑狼疮肾纤维化干预机制的研究
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(1. 云南中医药大学,云南 昆明 650500;2. 楚雄州中医医院,云南 楚雄 675099;3. 保山中医药高等专科学校,云南 保山 678000)

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基金项目: 云南省科技厅-云南中医药大学应用基础研究联合专项资金项目[2018FF001(-037)];云南省教育厅治疗痹症中医中药工程研究中心资助项目(云教发〔2022〕70号)


Study on the Intervention Mechanism of Ziyinqingrehuoxue Formula Combined with Glucocorticoid Against Renal Fibrosis in Systemic Lupus Erythematosus
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(1. Yunnan University of Chinese Medicine, Kunming 650500, China; 2. Chuxiong Prefecture Hospital of Traditional Chinese Medicine, Chuxiong 675099, China;3. Baoshan College of Traditional Chinese Medicine, Baoshan 678000, China)

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    摘要:

    目的 研究滋阴清热活血方联合激素对MRL/lpr狼疮小鼠肾纤维化的影响,揭示该方抗狼疮肾纤维化效应机制。方法 将MRL/lpr自发性狼疮小鼠按尿蛋白水平平均分为5组:模型组、醋酸泼尼松组、滋阴清热活血方组、滋阴清热活血方加激素全量组、滋阴清热活血方加激素半量组。药物干预4周后,H&E染色和Masson染色分别检测狼疮小鼠肾组织病理学改变和肾纤维变化情况;免疫荧光法检测肾脏IgG免疫复合物沉积;Western blot法测定模型肾脏Smad4和Smad7蛋白表达水平。结果 与模型组比较,各治疗组小鼠肾脏病理损伤显著减少;Masson染色结果显示,与模型组比较,滋阴清热活血方组和醋酸泼尼松组胶原沉积减少;与滋阴清热活血方组比较,滋阴清热活血方加激素全量组和滋阴清热活血方加激素半剂量组胶原沉积面积均减少。免疫荧光染色检测结果显示,与模型组比较,滋阴清热活血方组和醋酸泼尼松组肾脏免疫复合物沉积减弱;与滋阴清热活血方组比较,滋阴清热活血方加激素全量和半量组肾脏IgG免疫复合物荧光显著减少。与模型组比较,滋阴清热活血方组、滋阴清热活血方加激素全量组和激素半剂量组肾组织Smad4蛋白显著下降;滋阴清热活血方组、滋阴清热活血方加激素全量组和激素半剂量组肾组织Smad7显著增加,其中,中药联合激素的2组尤为显著。结论 滋阴清热活血方联合激素对MRL/lpr小鼠肾组织肾纤维化具有较好的改善作用,其作用机制可能与抑制Smad4表达、上调Smad7蛋白表达有关,该研究为临床滋阴清热活血方联合激素治疗系统红斑狼疮肾纤维化提供了生物学依据。

    Abstract:

    Objective To investigate therapeutic potential of ziyinqingrehuoxue formula(ZYQRHXF) combined the glucocorticoid in MRL/lpr lupus mice, and to reveal the mechanism of ZYQRHXF on anti-lupus renal fibrosis. Methods Mice with MRL/lpr spontaneous lupus were divided into 5 groups according to the level of proteinuria, glucocorticoid group, ZYQRHXF group, ZYQRHXF and glucocorticoid full volume group and ZYQRHXF and half-dose group, respectively. After 4 weeks of drug intervention, histopathological damage of kidney and the deposition of IgG immune complexes were assessed by H&E staining and immunofluorescence, respectively. Kidney fibrosis was detected by Masson staining; the protein expression levels of Smad4 and Smad7 in the kidneys were detected by Western blot. Results Compared with the model group, the pathological damage of kidney in treatment group mice was significantly reduced; Masson staining results showed that collagen deposited were significantly decreased in prednisone acetate group, ZYQRHXF and Glucocorticoid full volume group and ZYQRHXF half-dose group when compared to the model group and ZYQRHXF group. The deposition of renal IgG immune complex were reduced in the ZYQRHXF and glucocorticoid full volume group and half-dose group. Compared with the model group, Smad4 protein was decreased significantly in the ZYQRHXF group, glucocorticoid full volume group and half-dose group. While Smad7 in the ZYQRHXF group, glucocorticoid full volume group and half-dose group was dramatic increased, especially in both the traditional Chinese medicine combined the glucocorticoid groups. Conclusion The ZYQRHXF combined the glucocorticoid could ameliorated renal fibrosis in MRL/lpr mice. Its mechanism of action was related to the inhibition of Smad4 expression and the up-regulation of Smad7 protein expression. This study provides a biological basis for clinical treatment of ZYQRHXF in renal fibrosis of lupus erythematosus.

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  • 收稿日期:2022-11-08
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  • 在线发布日期: 2023-03-07
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