寒痉汤及其拆方对寒凝证高血压大鼠血清炎症因子、血管内皮功能及纤维化的影响

doi:10.6040/j.issn.1671-7554.0.2022.0380

摘要/Abstract

摘要： 目的观察比较寒痉汤及其拆方对寒凝证型高血压大鼠的降压效果以及对其血清炎症因子、血管内皮功能、血管胶原纤维的影响,探究寒痉汤发挥降压作用的相关机制。方法 6~8周龄雄性SD大白鼠40只随机分为正常组、模型组、寒痉汤组、通阳拆方组和通络拆方组,每组8只。正常组给予普通饲料,其他4组给予含 8% 氯化钠高盐饲料,同时每天置于(-15±2)℃冰柜中3 h,持续6周,给药2周,正常组、模型组每天给予等体积生理盐水灌胃。每周用大鼠无创血压分析系统测量血压2次,取平均值。釆用苏木精-伊红和马松(Masson)染色观察各组大鼠主动脉病理形态;采用ELISA法检测血清白介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)、白介素-6(IL-6)、内皮素-1(ET-1)含量,采用比色法检测血清一氧化氮(NO)含量;免疫组化(IHC)检测各组大鼠主动脉中内皮型一氧化氮合成酶(eNOS)、Ⅰ型胶原蛋白(ColI)、转化生长因子-β1(TGF-β1)的表达水平。结果 (1)与正常组比较,模型组血压升高,主动脉内膜不完整、中膜增厚、细胞排列紊乱,血清TNF-α、IL-1β、IL-6、ET-1升高、NO减少,胶原纤维增多(P<0.01);免疫组化显示主动脉eNOS表达降低,ColI、TGF-β1表达升高(P<0.01)。(2)与模型组比较,寒痉汤组、通阳拆方组和通络拆方组血压降低(P<0.01,P<0.05),主动脉结构有不同程度改善,血清 NO升高(P<0.05),其中寒痉汤组TNF-α、IL-1β、ET-1和IL-6降低(P<0.01,P<0.05),通阳拆方组TNF-α、IL-1β、IL-6降低(P<0.05),通络拆方组ET-1、TNF-α降低(P<0.05);IHC显示3个治疗组主动脉ColI表达减少、eNOS表达升高(P<0.05),寒痉汤组、通阳拆方组TGF-β1表达降低(P<0.01,P<0.05);Masson染色显示3个治疗组主动脉胶原沉积减少(P<0.01)。(3)与寒痉汤组比较,通阳拆方组、通络拆方组的降压效果和对主动脉胶原沉积的改善差异有统计学意义(P<0.05,P<0.01)。结论寒痉汤可减少血清IL-1β、TNF-α、IL-6炎症因子,改善内皮功能,减少血管胶原蛋白沉积,改善主动脉纤维化,可能是其发挥降压作用的相关机制。其通阳散寒成分主要发挥减少炎症因子的作用,而通络止痉成分主要是通过调节NO/ET-1水平来协同发挥其降压和保护血管的作用。

关键词: 寒痉汤, 寒凝证高血压, 主动脉纤维化, 炎症因子, 内皮功能

Abstract: Objective To observe and compare the antihypertensive effects of Hanjingtang and its disassembled prescriptions on hypertensive rats with cold coagulation syndrome and the influence on serum inflammatory factors, vascular endothelial function and vascular collagen fiber, so that to explore the antihypertensive mechanism. Methods A total of 40 male SD rats aged 6 to 8 weeks were randomly divided into control group, model group, Hanjingtang group, Tongyang group and Tongluo group, with 8 rats in each group. The control group was fed with ordinary diet, and the other groups were given diet with 8% sodium chloride and high salt, and placed in a freezer at(-15±2)℃ for 3 hours every day for 6 weeks, followed by intragastric administration for 2 weeks. The control group and model group were given equal volume of normal saline. The blood pressure was measured twice a week with the rat noninvasive blood pressure analysis system, and the average value was taken. The changes of aortic pathological morphology and collagen deposition were observed with HE and Masson staining. The IL-1β, TNF- α and ET-1 were detected with ELISA, and NO was detected with colorimetry. The expressions of eNOS, type I collagen(ColI)and TGF-β1 in aortic tissue were detected with IHC. Results (1) Compared with the control group, the model group had raised blood pressure(P<0.01); incomplete aortic intima; thicker intima-media; disordered cell arranging; increased serum TNF-α, IL-1β, IL-6 and ET-1; decreased NO; increased collagen fibers. Meanwhile, IHC showed that eNOS expression decreased, but ColI and TGF-β1 expressions increased in aorta(P<0.01). (2) Compared with the model group, the Hanjingtang, Tongyang and Tongluo groups had decreased blood pressure(P<0.01, P<0.05); improved aortic structure at different levels; increased serum NO(P<0.05); Hanjingtang group had decreased TNF-α, IL-1β, ET-1 and IL-6(P<0.01, P<0.05); Tongyang group had decreased TNF-α, IL-1β and IL-6(P<0.05); Tongluo group had decreased ET-1 and TNF-α(P<0.05). IHC showed that the expression of ColI in aorta decreased but eNOS increased in the three treatment groups(P<0.05). The expression of TGF-β1 decreased in Hanjingtang and Tongyang groups(P<0.01, P<0.05); aortic collagen fibers decreased in the three treatment groups(P<0.01). (3) The antihypertensive effects and improvement of aortic collagen deposition were better in the Tongyang and Tongluo groups than those in the Hanjingtang group(P<0.05, P<0.01). Conclusion Hanjingtang can significantly reduce the serum inflammatory factors of IL-1β, TNF-α and IL-6, improve endothelial function, reduce collagen deposition in blood vessels, improve aortic fibrosis, partially explaining the antihypertensive mechanism of Hanjingtang. The Tongyang ingredients can mainly reduce inflammatory factors, while Tongluo ingredients can regulate NO / ET-1 levels to play a synergistic role.

Key words: Hanjingtang, Hypertension with cold syndrome, Aortic fibrosis, Inflammation factors, Endothelial dysfunction

中图分类号:

R285.5

邬雨洁,张明泉,纪永利,赵璐,王越,陈沙沙. 寒痉汤及其拆方对寒凝证高血压大鼠血清炎症因子、血管内皮功能及纤维化的影响[J]. 山东大学学报 (医学版), 2022, 60(6): 10-18.

WU Yujie, ZHANG Mingquan, JI Yongli, ZHAO Lu, WANG Yue, CHEN Shasha. Effects of Hanjingtang and its disassembled prescriptions on serum inflammatory factors, vascular endothelial function and fibrosis in hypertensive rats with cold coagulation syndrome[J]. Journal of Shandong University (Health Sciences), 2022, 60(6): 10-18.

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