紫苏叶对感染性肺损伤作用及作用机制

The effect and mechanism of perilla leaves on infectious lung injury

关键词:  紫苏叶  感染性肺损伤  细胞因子风暴  c-Myc

Key words:  perilla leaves  infectious lung injury  cytokine storm  c-Myc

摘要:

摘要：目的 本文旨在探讨紫苏叶（Perilla frutescens (L.) Britt.）对LPS导致的炎性肺损伤的药效及作用机制。方法 实验分为空白对照组、模型组、阳性药组及紫苏叶给药组。以BEAS-2B细胞为宿主细胞，利用LPS（5 μg·mL?1）建立细胞炎性损伤模型。通过CCK-8、LDH法及流式细胞术评价细胞损伤情况，并通过酶联免疫吸附实验法检测炎症因子的表达；采用LPS（5 mg·kg?1）建立小鼠肺部炎性损伤模型，并检测肺泡灌洗液（BALF）中总细胞数量与总蛋白浓度水平，评价其肺部损伤状况。利用网络药理学预测紫苏叶提取物治疗感染性肺损伤的潜在作用靶点，并通过Western-blot验证其作用机制。结果 体外实验结果显示，紫苏叶可以改善细胞生长情况、提高细胞活力、抑制细胞凋亡及改善细胞损伤情况，调控细胞损伤后炎症因子白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）的水平；在体内实验结果证实紫苏叶能够减少BALF中总细胞数量，降低BALF中总蛋白浓度水平，减轻小鼠肺组织细胞凋亡。Western-blot结果提示紫苏叶能够抑制c-Myc、p-MEK1/2、SRF的表达。结论 紫苏叶能有效抑制LPS引起的BEAS-2B细胞损伤，降低炎症因子水平，减少BALF中总细胞数量与总蛋白浓度，改善肺组织损伤。其机制与紫苏叶调控c-Myc、p-MEK1/2、SRF蛋白表达相关。

Abstract:

ABSTRACT: OBJECTIVE This study was to investigate the efficacy and mechanism of action of perilla leaves(Perilla frutescens (L.) Britt.) on LPS-induced inflammatory lung injury. METHODS Experimental groups were control, model, positive drug, and perilla leaves extract group. BEAS-2B cells were used as host cells to establish a cell inflammatory injury model using LPS (5 μg·mL?1). Cell damage was evaluated by CCK-8, LDH assay, and flow cytometry, and the expression of inflammatory factors was detected by ELISA. A mouse lung inflammatory injury model was established using LPS (5 mg·kg?1), and the total cell number and total protein concentration in bronchoalveolar lavage fluid (BALF) were measured to evaluate lung injury. Finally, Network pharmacology was used to predict potential targets of perilla leaves extract in the treatment of infectious pneumonia, and the mechanism of action was validated by Western blot. RESULTS Perilla leaves improved cell growth, increased cell viability, inhibited cell apoptosis, and improved cell damage in vitro, regulating the levels of inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) after cell damage; In vivo experiments confirmed that perilla leaves reduced the total cell number in BALF, decreased the total protein concentration in BALF, and alleviated lung tissue cell apoptosis. Western-blot suggesting that perilla leaves could inhibit the expression of c-Myc, p-MEK1/2, and SRF. CONCLUSION Perilla leaves effectively inhibited LPS-induced BEAS-2B cell damage, reduced inflammatory factor levels, decreased the total cell number and total protein concentration in BALF, and improved lung tissue damage. The mechanism of action may be related to the regulation of c-Myc, p-MEK1/2, and SRF protein expression by perilla leaves.