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补肾和脉颗粒通过调节Th17细胞/Tregs平衡改善老年自发性高血压大鼠肾损伤的机制研究

Study on the Mechanism of Bu-Shen-He-Mai Granules in Improving Renal Damage of Ageing Spontaneously Hypertensive Rats by Regulating Th17 Cell/Tregs Balance.

作者信息

Zhang Peng, Song Xu-Yu, Li Wen, Wei Jian-Liang, Cui Yan-Jun, Qi Ying-Zi, Chen Xiu-Bao, Jiang Yue-Hua, Yang Chuan-Hua

机构信息

First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan 250014, China.

Department of Cardiovascular, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250014, China.

出版信息

Evid Based Complement Alternat Med. 2022 Apr 23;2022:8315503. doi: 10.1155/2022/8315503. eCollection 2022.

Abstract

METHODS

Blood pressure and urine biochemical indices were recorded. Renal blood flow was evaluated by renal ultrasonography. Transmission electron microscopy (TEM) and HE staining were used to assess kidney and spleen morphology. Renal fibrosis was assessed using Masson staining. Serum levels of IL-6, IL-10, and IL-17A were measured using ELISAs. The density of ROR and Foxp3 in the spleen was observed by immunofluorescence staining. The levels of Th17 cells and Tregs in blood were detected via flow cytometry. Transcriptome sequencing was performed to screen the targets of BSHM granules in hypertensive kidneys.

RESULTS

BSHM granules decreased SBP by 21.2 mm·Hg and DBP by 8.8 mm·Hg in ageing SHRs ( < 0.05), decreased the levels of urine mALB, 2-Mg, and NAG ( < 0.01), and improved renal blood flow and arteriosclerosis. BSHM granules increased IL-10 expression ( < 0.05) while decreasing IL-6 ( < 0.01) and IL-17A ( < 0.05) levels. BSHM granules improved Foxp3 density and the number of Tregs ( < 0.01) and reduced RORt density and the number of Th17 cells ( < 0.01). Transcriptome sequencing identified 747 differentially expressed (DE) mRNAs in kidneys after BSHM treatment. GO analysis suggested that BSHM granules act through immunoregulation.

CONCLUSIONS

BSHM granules attenuated hypertensive renal damage in ageing SHRs, by significantly increasing Tregs and decreasing Th17 cells.

摘要

方法

记录血压和尿液生化指标。通过肾脏超声评估肾血流量。采用透射电子显微镜(TEM)和苏木精-伊红(HE)染色评估肾脏和脾脏形态。使用Masson染色评估肾纤维化。采用酶联免疫吸附测定(ELISA)法检测血清白细胞介素-6(IL-6)、白细胞介素-10(IL-10)和白细胞介素-17A(IL-17A)水平。通过免疫荧光染色观察脾脏中维甲酸相关孤儿受体(ROR)和叉头框蛋白3(Foxp3)的密度。通过流式细胞术检测血液中辅助性T细胞17(Th17)细胞和调节性T细胞(Tregs)的水平。进行转录组测序以筛选补肾活血颗粒在高血压肾脏中的作用靶点。

结果

补肾活血颗粒使老龄自发性高血压大鼠(SHRs)的收缩压降低21.2 mmHg,舒张压降低8.8 mmHg(P<0.05),降低尿微量白蛋白(mALB)、β2微球蛋白(2-Mg)和N-乙酰-β-D-氨基葡萄糖苷酶(NAG)水平(P<0.01),并改善肾血流量和动脉硬化。补肾活血颗粒增加IL-10表达(P<0.05),同时降低IL-6(P<0.01)和IL-17A水平(P<0.05)。补肾活血颗粒改善Foxp3密度和Tregs数量(P<0.01),降低RORγt密度和Th17细胞数量(P<0.01)。转录组测序鉴定出补肾活血颗粒治疗后肾脏中有747个差异表达的mRNA。基因本体(GO)分析表明,补肾活血颗粒通过免疫调节发挥作用。

结论

补肾活血颗粒通过显著增加Tregs并减少Th17细胞,减轻老龄SHRs的高血压肾损伤。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aaf4/9056229/a7e67b97c2ee/ECAM2022-8315503.001.jpg

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