Tengdan Capsule Prevents Hypertensive Kidney Damage in SHR by Inhibiting Periostin-Mediated Renal Fibrosis.
Tengdan Capsule Prevents Hypertensive Kidney Damage in SHR by Inhibiting Periostin-Mediated Renal Fibrosis.
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腾丹胶囊通过抑制骨膜素介导的肾纤维化预防 SHR 高血压肾损伤
DOI：
10.3389/fphar.2021.638298
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发表时间：
2021
影响因子：
5.6
通讯作者：
Zhu Y
中科院分区：
文献类型：
--
作者：
Du X;Tao Q;Du H;Zhao Z;Dong Y;He S;Shao R;Wang Y;Han W;Wang X;Zhu Y
背景：高血压性肾损害是一种严重而复杂的疾病，传统降压药尚未有效治疗。腾丹胶囊（TDC）是中国fda批准的治疗高血压的复方草药；然而，其化学基础和药理功效尚未在临床前环境中得到充分研究。方法：采用高效液相色谱法对从超纯水中提取的TDC的主要化学成分进行鉴定和定量。将成年自发性高血压大鼠（SHR）和年龄/性别匹配的Wistar京都正常血压大鼠（WKY）分别给予TDC、氯沙坦或生理盐水治疗1个月，同时用尾袖血压系统监测血压（BP）。测定尿肌酐（CRE）、血尿素氮（BUN）等生化指标。肾组织切片（H&E）、Masson染色观察TDC对SHR肾脏的病理影响。在TDC治疗后，通过基于TDC的定量蛋白质组学分析，鉴定了SHR肾脏中差异表达的蛋白，可能为TDC的作用靶点和可能的机制提供线索。此外，我们还进行了Western blot分析、RT-qPCR和ELISA分析来进一步验证蛋白质组学的发现。最后，利用人肾HEK293细胞建立两种不同的体外肾损伤模型；探讨了TDC保护肾脏的分子机制。结果：采用高效液相色谱法从三七皂苷水溶性提取物中分离鉴定出7个化合物，分别为三七皂苷R1、人参皂苷RG1、人参皂苷Re、人参皂苷Rb1、丹参素钠、原儿茶醛、丹酚酸B。大鼠体内研究表明，TDC可有效降低SHR的BP、BUN和CRE水平，减轻肾纤维化，改善肾脏损伤。蛋白质组学和随后的生物信息学分析表明，骨膜蛋白介导的炎症反应和TGFβ/Smad信号通路蛋白与TDC对大鼠肾脏的治疗效果密切相关。Western blot和RT-qPCR分析显示，与未处理的SHR相比，TDC显著下调肾组织中骨膜蛋白mRNA和蛋白的表达。此外，TDC治疗后SHR肾组织中TGF-β和COL1A1 mRNA水平也下降。体外研究表明，低至中剂量TDC可下调HEK293细胞损伤模型中骨膜蛋白的表达。此外，中至高剂量TDC可显著抑制tgf β1诱导的HEK293细胞纤维化中的胶原沉积。结论：对复方中药腾丹胶囊的主要成分进行了鉴定和定量。TDC有效降低血压，防止SHR患者高血压引起的肾脏损害。在体内和体外实验中，TDC通过调节肾脏TGF-β/Smad信号通路阻断骨膜素。预防骨膜蛋白介导的肾纤维化和炎症可能是治疗高血压肾损伤的一种有希望的策略。
BACKGROUND: Hypertension-induced renal damage is a serious and complex condition that has not been effectively treated by conventional blood pressure-lowering drugs. Tengdan capsule (TDC) is a China FDA-approved compound herbal medicine for treating hypertension; however, its chemical basis and pharmacological efficacy have not been fully investigated in a preclinical setting. METHODS: High-performance liquid chromatography (HPLC) was used to identify and quantify the major chemical components of TDC extracted from ultrapure water. Adult spontaneously hypertensive rats (SHR) and age/sex-matched Wistar Kyoto normotensive rats (WKY) were both treated with TDC, losartan, or saline for one month, and their blood pressure (BP) was monitored at the same time by tail-cuff BP system. Biochemical indexes such as urine creatinine (CRE) and blood urea nitrogen (BUN) were determined. Kidney tissue sections were examined with (H&E), and Masson staining to evaluate the pathological effect of TDC on SHR’s kidneys. After TDC treatment, the differentially expressed proteins in the kidneys of SHR were identified by the TMT-based quantitative proteomics analysis, which may provide the targets and possible mechanisms of TDC action. In addition, Western blot analysis, RT-qPCR, and ELISA assays were carried out to further verify the proteomics findings. Finally, two different models involving in vitro renal injuries were established using human kidney HEK293 cells; and the molecular mechanism of TDC kidney protection was demonstrated. RESULTS: Seven chemical compounds, namely Notoginsenoside R1, Ginsenoside RG1, Ginsenoside Re, Ginsenoside Rb1, Sodium Danshensu, Protocatechualdehyde, and Salvianolic acid B, were identified and quantified from the water-soluble extracts of TDC by HPLC. In vivo study using rats showed that TDC effectively reduced BP, BUN, and CRE levels and attenuated renal fibrosis in SHR, and ameliorated damage to the kidneys. Proteomics and subsequent bioinformatics analyses indicated that periostin-mediated inflammatory response and TGFβ/Smad signaling pathway proteins were closely related to the therapeutic effect of TDC in rat kidneys. Western blot analysis and RT-qPCR showed that TDC markedly downregulated the mRNA and protein expression of periostin in renal tissues compared to the untreated SHR. In addition, TGF-β and COL1A1 mRNA levels also decreased in SHR renal tissues following TDC treatment. In vitro studies showed that low to medium doses of TDC down-regulated the expression of periostin in the injury model of HEK293 cell. In addition, medium to high doses of TDC significantly inhibited collagen deposition in TGFβ1-induced HEK293 cell fibrosis. CONCLUSIONS: Major components from the compound herbal medicine Tengdan Capsule are identified and quantified. TDC effectively lowers blood pressure and protects against renal damage caused by hypertension in SHR. Mechanistically, TDC blocks periostin by regulating the TGF-β/Smad signaling pathway in the kidney, both in vivo and in vitro. Preventing periostin-mediated renal fibrosis and inflammation might be a promising strategy for treating a hypertensive renal injury.
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影响因子：
3.2
作者：
Farris AB;Colvin RB
通讯作者：
Colvin RB
影响因子：
--
作者：
Chen H;Yin J;Deng Y;Yang M;Xu L;Teng F;Li D;Cheng Y;Liu S;Wang D;Zhang T;Wu W;Liu X;Guan S;Jiang B;Guo D
通讯作者：
Guo D
影响因子：
15.9
作者：
KAGAMI, S;BORDER, WA;NOBLE, NA
通讯作者：
NOBLE, NA
影响因子：
2
作者：
He, Yu L.;Wen, Jian G.;Bauer, Stuart B.
通讯作者：
Bauer, Stuart B.
DOI：
10.1152/ajprenal.00273.2016
发表时间：
2016-12-01
影响因子：
4.2
作者：
Crowley, Steven D.;Jeffs, Alexander D.
通讯作者：
Jeffs, Alexander D.