基于“LncRNA MALAT1-巨噬细胞焦亡”新视角探讨加味四妙勇安汤促进大鼠糖尿病足溃疡愈合机制  

Exploring the Mechanism of Modified Simiao Yong’an Decoction for Promoting Diabetic Foot Ulcers Healing in Rats Based on A New Perspective of“LncRNA MALAT1-Macrophage Pyroptosis”

作　　者：王庆美 徐鹏 付媛媛 潘银燕 WANG Qing-Mei;XU Peng;FU Yuan-Yuan;PAN Yin-Yan(Shandong Provincial Third Hospital,Jinan 250031 Shandong,China;The 960th Hospital of PLA,Jinan 250031 Shandong,China)

机构地区：[1]山东省立第三医院,山东济南250031 [2]解放军第九六○医院,山东济南250031

出　　处：《广州中医药大学学报》2023年第2期413-420,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:82004369)。

摘　　要：【目的】探讨加味四妙勇安汤通过长链非编码RNA肺腺癌转移相关转录本1(LncRNA MALAT1)-巨噬细胞焦亡轴对大鼠糖尿病足溃疡的治疗作用。【方法】将40只SD大鼠随机分为正常组、模型组、siLncRNA MALAT1(慢病毒质粒)组、加味四妙勇安汤组,每组10只。除正常组,其余各组大鼠构建糖尿病足溃疡模型。成功造模后,进行相应干预。观察大鼠治疗前及治疗4、8周后足溃疡创面愈合情况,计算大鼠治疗前后创面愈合面积百分比。干预结束后,采用酶联免疫吸附分析(ELISA)检测大鼠血清白细胞介素(IL)-18、IL-1β水平,免疫荧光化学染色法检测足溃疡创面组织M2巨噬细胞标志蛋白(CD206)、凋亡相关斑点样蛋白(ASC)、Nod样受体家族含pyrin结构域蛋白3(NLRP3)表达,实时聚合酶链反应(RT-PCR)法检测足溃疡创面组织LncRNA MALAT1表达,Western Blot法检测足溃疡创面组织细胞焦亡关键蛋白消皮素D(GSDMD)、NLRP3、proCaspase-1、核转录因子kappaB(NF-κB)p65等的蛋白表达。【结果】(1)与模型组比较,siLncRNA MALAT1组、加味四妙勇安汤组足溃疡创面面积减小。(2)与正常组比较,模型组血清IL-18、IL-1β水平,足溃疡创面组织LncRNA MALAT1表达水平,ASC、NLRP3、GSDMD、pro-Caspase-1、NF-κB p65蛋白表达水平升高,CD206蛋白表达水平降低(均P<0.05);与模型组比较,siLncRNA MALAT1组、加味四妙勇安汤组血清IL-18、IL-1β水平,足溃疡创面组织LncRNA MALAT1水平,ASC、NLRP3、GSDMD、pro-Caspase-1、NF-κB p65蛋白表达水平降低,CD206蛋白表达水平升高(均P<0.05)。加味四妙勇安汤组上述各指标与siLncRNA MALAT1组比较,差异均无统计学意义(P>0.05)。【结论】加味四妙勇安汤可通过下调LncRNA MALAT1抑制巨噬细胞焦亡,从而促进大鼠糖尿病足溃疡愈合。Objective To investigate the therapeutic effect of modified Simiao Yong’an Decoction on diabetic foot ulcers in rats via long-stranded non-coding RNA lung adenocarcinoma metastasis-associated transcript 1(LncRNA MALAT1)-macrophage pyroptosis axis. Methods Forty SD rats were randomly divided into the normal group,the model group, the siLncRNA MALAT1(lentiviral plasmid)group, and modified Simiao Yong’an Decoction group, with 10 rats in each group. Except for the normal group, the rat model of diabetic foot ulcer was constructed in all groups. After successful modeling,the corresponding interventions were carried out. The healing of the foot ulcers was observed before treatment and after 4 and 8 weeks of treatment,and the percentage of healed area was calculated before and after treatment. After the intervention,serum levels of interleukin(IL)-18 and IL-1β were measured by enzyme-linked immunosorbent assay(ELISA)and the expression of M2 macrophage marker protein(CD206), apoptosis-associated spot-like protein(ASC)and Nod-like receptor family pyrincontaining protein 3(NLRP3)in the healing tissue of foot ulcers were measured by immunofluorescence chemical staining. The expression of LncRNA MALAT1 in the healing tissue of foot ulcers was detected by polymerase chain reaction(RT-PCR), and the protein expressions of gasdermin D(GSDMD), NLRP3, pro-Caspase-1 and nuclear transcription factor kappaB(NF-κB)p65 in the healing tissue of foot ulcers were detected by Western Blot.Results(1)The wound area in the siLncRNA MALAT1 group and the modified Simiao Yong’an Decoction group was reduced compared with the model group.(2)Compared with the normal group,the serum levels of IL-18,IL-1β,the LncRNA MALAT1 expression level in the healing tissue of foot ulcers,and the protein expression levels of ASC, NLRP3, GSDMD, pro-Caspase-1 and NF-κB p65 were increased and CD206 protein was decreased in the model group(all P<0.05);compared with the model group,the serum levels of IL-18,IL-1β,the LncRNA MALAT1 expression level in the he

关 键 词：加味四妙勇安汤 糖尿病足 溃疡愈合 LncRNA MALAT1 巨噬细胞 细胞焦亡 大鼠 

分 类 号：R285.5[医药卫生—中药学]