火坦解毒通络汤对 H 型高血压大鼠的降压作用及机制
Summary
在这里，我们提出了一种诱导 H 型高血压的方案，并评估了胃内给药的火坦解毒通络汤 （HTJDTLD） 的抗高血压效果。在 H 型高血压大鼠中，HTJDTLD 具有有效的降压作用，可能与抑制内质网 （ER） 应激诱导的细胞凋亡通路激活有关。
Abstract
H 型高血压是一种以血浆同型半胱氨酸 （Hcy） 水平升高为特征的特殊高血压，已成为全球面临的重大公共卫生挑战。本研究探讨了常用于治疗血管狭窄的高效中药方剂火坦解毒通络汤 （HTJDTLD） 的降压作用和潜在机制。蛋氨酸诱导大鼠 H 型高血压，HTJDTLD 经胃内给药。然后，通过无创大鼠尾部测压法测量大鼠尾动脉的收缩压和舒张压。通过苏木精-伊红 （HE） 染色进行主动脉的组织学评估。采用酶联免疫吸附试验 （ELISA） 检测 Hcy 水平，定量逆转录酶聚合酶链反应 （qRT-PCR） 和蛋白质印迹法测定葡萄糖调节蛋白 78 （GRP78） 、肿瘤坏死因子 （TNF） 受体相关因子 2 （TRAF2） 、c-Jun N 末端激酶 （JNK） 和 caspase-3 的 mRNA 和蛋白水平。结果显示，蛋氨酸治疗后 HTJDTLD 可显著降低血压，减轻组织病理学病变，降低 Hcy 水平。此外，HTJDTLD 显著抑制 GRP78 、 JNK 、 TRAF2 和 caspase 3 的基因和蛋白表达，这些基因主要参与内质网 （ER） 应激诱导的细胞凋亡途径。总体而言，结果表明 HTJDTLD 对 H 型高血压大鼠具有有效的降压作用，并揭示了与抑制 ER 应激诱导的细胞凋亡通路激活相关的降压机制。
Introduction
高血压是心脏病发作、中风和肾功能衰竭的主要风险因素，已成为影响全球 10 亿人的重大公共卫生挑战1。同型半胱氨酸 （Hcy） 是一种含硫醇基团的氨基酸，是蛋氨酸代谢的重要代谢中介。血浆 Hcy 水平升高的高血压定义为 H 型高血压，这可能是中风等心脑血管疾病发生和复发的重要危险因素 2,3。最近的研究报告指出，H 型高血压共存可能会加重心脑血管疾病的副作用4。值得注意的是，中国 75% 的 H 型高血压患者患有原发性高血压，严重影响生活质量5。目前，H型高血压的治疗主要包括西医。但可能会引起一定的不良反应和依从性差，不能再满足H型高血压综合管理的需要。
中医 （TCM） 在中国是一种独特的资源，已有 2,000 多年的历史。由于西医对高血压控制的需求未得到满足，临床医生已经开始考虑中医在预防和治疗 H 型高血压中的潜在作用6。火坛解毒通络汤 （HTJDTLD） 是由邓悦教授根据其广泛的临床专业知识配制的中药配方7.经过 20 多年的临床应用，HTJDTLD 在治疗心脑血管疾病方面显示出显著的疗效1。然而，HTJDTLD 是否对 H 型高血压有治疗作用尚未见报道。因此，我们旨在探讨 HTJDTLD 对 H 型高血压大鼠的降压作用和特异性机制，并确定治疗 H 型高血压的潜在治疗药物。
Protocol
所有动物实验均经长春中医药大学机构动物护理和使用委员会 （IACUC） 批准。材料表列出了这些材料。 1. 动物和治疗 将总共 50 只成年自发性高血压大鼠 （SHR） （雄性，50 天龄） 随机分为 5 组，包括对照组 （CON）、蛋氨酸 （MET）、MET + HTJDTLD + 马来酸依那普利 （EM）、MET + EM 和 MET + HTJDTLD 组。注意：实验大鼠被安置在受控环境中，旨在?…
Representative Results
如 表 4 和 表 5 所示，MET 组收缩压 （SBP） 和舒张压 （DBP） 在 1 至 4 周内显著高于 CON 组。HTJDTLD 治疗后，大鼠 SBP 和 DBP 显著低于 MET 组。值得注意的是，HTJDTLD 和 EM 联合使用比单独使用 HTJDTLD 治疗具有更强的降压效果。 经 HE 染色和 Masson 三色染色，主动脉血管壁子宫内膜不完整且不光滑，中层明显增厚。与 CON 组相比，MET 组出现平滑肌细胞增殖和?…
Discussion
高血压是最常见的心血管疾病之一，影响三分之一的成年人口，并增加中风、冠心病以及心肾衰竭的风险8。H 型高血压是一种特殊类型的高血压，是指原发性高血压和同型半胱氨酸水平升高同时发生，多年来引起了广泛关注9。近年来，口服 EM 联合降压药在控制血压和降低血液 Hcy 水平方面显示出更好的疗效，但仍存在许多禁忌症、副作用、耐药性以及头痛、头…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作得到了吉林省国家自然科学基金 （no.YDZJ202301ZYTS189）。
Materials
| 1st Strand cDNA Synthesis SuperMix for qPCR | Yeasen,China | 11149ES | cDNA synthesis kit |
| Anti-beta-actin antibody | Bioss, China | bs-0061R | |
| Anti-caspase-3 antibody | Bioss, China | bs-0081R | |
| Anti-GPR78 antibody | Abcam, USA | ab108513 | |
| Anti-JNK antibody | Abcam, USA | ab76572 | |
| Anti-p-JNK antibody | Bioss, China | bsm-52462R | |
| Anti-rabbit IgG antibody | Bioss, China | bs-0295G-HRP | |
| Anti-TRAF2 antibody | Bioss, China | bs-22372R | |
| Bio-Rad CFX96 Touch system | Bio-Rad | CFX96 | real-time PCR detection system |
| ECL Western Blot Substrates | Merck, MA, USA | WBULP-10ML | |
| Enalapril maleate folic acid tablets | Yangzijiang Pharmaceutical Company, China | 20040991 | |
| FastStart SYBR Green Master | Sigma | FSSGMMRO | |
| Fructus Trichosanthis | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Hirudo | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| intelligent noninvasive sphygmomanometer | Beijing Softron Biotechnology company | BP-2010A | |
| Lonicerae Japonicae Flos | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Methionine | Sigma, USA | M9500 | |
| Radix Angelicae Sinensis | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Radix et Rhizoma Glycyrrhizae | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Radix et Rhizoma Nardostachyos | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Radix et Rhizoma Rhodiolae Crenulatae | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Radix et Rhizoma Salviae Miltiorrhizae | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Radix Scrophulariae | The First Affiliated Hospital of Changchun University of Traditional Chinese Medicine, China | No catalog number | |
| Rat Hcy ELISA Kits | Shanghai Meimian Industrial Company, China | MM-0293R2 | |
| RIPA buffer | Shanghai Beyotime Biotechnology company | P0013B |
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Cite This Article
Shen, J., Hu, C., Li, Y., Shang, X., Deng, Y., Guo, J., Zhang, L., Wang, J., Zhang, W. The Antihypertensive Effects and Mechanisms of Huotan Jiedu Tongluo Decoction in Rats with H-Type Hypertension. J. Vis. Exp. (207), e65932, doi:10.3791/65932 (2024).