基于“心脾同治”理论探讨益气活血方调控冠心病气虚血瘀证大鼠炎症反应的机制
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1. 湖南中医药大学第一附属医院,长沙 410007;2. 湖南中医药大学,长沙 410208

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Inflammatory mechanism of Yiqi Huoxue recipe with qi deficiency and blood stasis syndrome of coronary heart disease based on the theory of simultaneous treatment of heart and spleen
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1. the First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China. 2. Hunan University of Chinese Medicine, Changsha 410208

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    摘要:

    目的 本研究拟探讨益气活血方调控冠心病气虚血瘀证大鼠免疫炎症的机制。 方法 30 只 SD 大鼠随机分为正常组、模型组、中药组。 模型组和中药组均采用“力竭+控食+冠状动脉左前降支结扎法”制备冠心病气虚血瘀证模型,造模周期为 22 d,中药组在造模第 8 天起予以益气活血方预处理 14 d。 于第 25 天取大鼠血清、脾、主动脉及缺血区心肌组织。 采用酶联免疫吸附测定法(ELISA)检测各组大鼠血清中 IL-35、IL-37 含量;HE 染色观察脾、心肌组织病理形态;Masson 染色观察主动脉病理形态;免疫组化法、Western Blot 分别检测脾、心、主动脉IL-35、IL-37 光密度表达及蛋白表达。 结果 (1)模型组较正常组血清中 IL-37、IL-35 显著降低(P< 0. 00001);中药组较模型组 IL-37、IL-35 均显著升高(P< 0. 00001)。 (2)HE 染色显示模型组心肌间质可见大量炎性细胞浸润;脾组织红髓区域变小,白髓可见边缘区(marginal zone,MZ)和动脉周围淋巴鞘之间淋巴小结大量增多。 中药组心肌组织炎性浸润较模型组明显减少,脾组织可见正常分布的脾小梁( trabecula,T)、血窦( splenic sinus,SS),且脾中央动脉(central antery,CA)形态规则。 Masson 染色显示模型组主动脉内膜增生,结构紊乱并存在泡沫细胞、脂质沉淀;中药组主动脉内膜变薄,纤维破损处可见修复痕迹。 (3)模型组大鼠心肌、主动脉、脾组织 IL-35、IL-37 平均光密度值及蛋白含量显著低于正常组(P< 0. 00001);中药组大鼠心肌、主动脉、脾组织 IL-35、IL-37 平均光密度及蛋白含量显著高于模型组(P< 0. 00001)。 结论 CHD 气虚血瘀证模型大鼠可为“心脾同治”理论提供模型支撑;益气活血方对 CHD 气虚血瘀证炎症状态有改善作用,其机制与上调 IL-35、IL-37 水平相关。

    Abstract:

    Objective The aim of this study was to explore the mechanism of Yiqi Huoxue recipe in regulating immune inflammatory response in rats with coronary heart disease syndrome of qi deficiency and blood stasis. Methods Thirty SD rats were randomly divided into control group, model group, and recipe group. The model of qi deficiency and blood stasis syndrome of coronary heart disease was prepared by exhaustion, food control, and ligation of the left anterior descending branch of the coronary artery in model group, and recipe group. recipe group was pretreated with Yiqi Huoxue Recipe for 14 days from day 8 of modeling. On day 25, serum, heart, spleen, and aorta were collected for cytokine quantification and histopathological examination. Results (1)Serum levels of IL-37 and IL-35 were significantly lower in model group, than control group ( P< 0. 00001 ). ( 2 ) After treatment, the serum levels of IL-37 and IL-35 were significantly higher in recipe group than model group(P< 0. 00001). HE staining showed a large degree of inflammatory cell infiltration in the myocardial stroma in model group. The red pulp area of spleen tissue was smaller, and a large number of lymph nodes between marginal zone ( MZ) and the periarterial lymph sheath were seen in white pulp. Hyperplasia, structural disorder, the presence of foam cells, and lipid deposition were seen in the aortic intima in model group. Inflammatory cell infiltration of myocardial tissue was significantly lower in recipe group than in the model group. The intima of the aorta was thin, and scars were seen at damaged fibers. A normal distribution of trabecula(T)and splenic sinus(SS) was seen in spleen tissue and central antery(CA) morphology was regular in recipe group. ( 3) The average optical densities and protein contents of IL-35 and IL-37 in the myocardium, aorta, and spleen of model group were significantly lower than those in control group(P< 0. 00001). The average optical densities and protein contents of IL-35 and IL-37 in the myocardium, aorta, and spleen of rats in recipe group were significantly higher than those in control group (P< 0. 00001). Conclusions CHD model rats with qi deficiency and blood stasis syndrome support the theory of simultaneous treatment of heart and spleen, Yiqi Huoxue recipe improves the inflammatory state of CHD with qi deficiency and blood stasis syndrome, and its mechanism is related to the upregulation of IL-35 and IL-37.

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王瑾茜,喻嵘,李薇,杨蕙,张田田,蔺晓源,胡国恒,陈亚,雷诗卉.基于“心脾同治”理论探讨益气活血方调控冠心病气虚血瘀证大鼠炎症反应的机制[J].中国实验动物学报,2022,30(7):935~941.

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  • 收稿日期:2022-05-12
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  • 在线发布日期: 2023-03-10
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