Ameliorative effects of traditional Chinese medicine formula NRICM101 on periodontal inflammation: In vitro and in vivo

Fang-Wei Lin , Yann-Jen Chen , Hui-Rong Cheng , Chi-Tsung Wu , Hsien-Chung Chiu
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Abstract

Introduction

Taiwan Chingguan Yihau (NRICM101) is a novel anti-inflammatory traditional Chinese medicine formula developed in 2020 for COVID-19 pandemic. This study aimed to evaluate the effect of NRICM101 on periodontal inflammation, using human gingival fibroblasts (HGFs) exposed to Porphyromonas gingivalis lipopolysaccharide (Pg LPS) and rat models of ligature-induced periodontitis.

Methods

In vitro, HGFs were stimulated with Pg LPS and NRICM101. Cell viability and the expression of pro-inflammatory cytokines, including interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and IL-1β were measured using MTS assay and reverse transcription quantitative polymerase chain reaction (RT-qPCR). In vivo, Sprague-Dawley rats were divided into control, ligature placement, and NRICM101 treatment groups. Ligature placement was performed at day one and oral administration of NRICM101 (50 or 100 mg/kg body weight/day) or saline began one day prior to ligation and the following days. On day 8, the rats were sacrificed, periodontal tissue specimens were analyzed using microcomputed tomography (Micro-CT) imaging, RT-qPCR, and immunohistochemical (IHC) staining to evaluate the effects of the treatments.

Results

NRICM101 reduced HGFs viability above 1.0 mg/mL, while Pg LPS caused similar effects at concentrations above 1.0 μg/mL. Co-treatment with Pg LPS (0.5–1.0 μg /mL) and NRICM101 (0.1–0.5 mg/mL) showed no cytotoxicity. In vivo, NRICM101 attenuated alveolar bone loss and significantly decreased IL-6, TNF-α, and partially IL-1β expression, as confirmed by micro-CT, RT-qPCR, and IHC analyses.

Discussion

This study demonstrates that NRICM101 exerts protective anti-inflammatory effects in both cellular and animal models of periodontitis. It suppressed pro-inflammatory cytokine expression in HGFs and mitigated alveolar bone loss in ligature-induced periodontitis rats. The effects on IL-6 and TNF-α were pronounced, while the partial reduction of IL-1β may reflect additional regulatory mechanisms such as inflammasome activation, which warrants further investigation. These findings indicate that NRICM101 reduces inflammation without cytotoxicity and supports its potential application in managing chronic periodontal inflammation.

Abstract Image

中药方剂NRICM101对牙周炎症的改善作用:体外和体内研究
台湾青观益口(NRICM101)是2020年针对新冠肺炎大流行研发的新型抗炎中药配方。本研究旨在通过暴露于牙龈卟啉单胞菌脂多糖(Pg LPS)的人牙龈成纤维细胞(HGFs)和结痂性牙周炎大鼠模型,评估NRICM101对牙周炎症的影响。方法体外用Pg LPS和NRICM101刺激hgf。采用MTS法和逆转录定量聚合酶链反应(RT-qPCR)检测细胞活力和促炎细胞因子,包括白细胞介素-6 (IL-6)、肿瘤坏死因子-α (TNF-α)和IL-1β的表达。在体内,Sprague-Dawley大鼠分为对照组、结扎组和NRICM101治疗组。在结扎的第一天进行结扎,并在结扎前一天和之后的几天开始口服NRICM101(50或100 mg/kg体重/天)或生理盐水。第8天处死大鼠,采用显微计算机断层扫描(Micro-CT)成像、RT-qPCR和免疫组化(IHC)染色对牙周组织标本进行分析,评价治疗效果。结果snricm101在1.0 mg/mL以上可降低hgf活力,Pg LPS在1.0 μg/mL以上可降低hgf活力。Pg LPS (0.5 ~ 1.0 μg /mL)与NRICM101 (0.1 ~ 0.5 mg/mL)共处理无细胞毒性。显微ct、RT-qPCR和免疫组化分析证实,在体内,NRICM101可减轻牙槽骨丢失,并显著降低IL-6、TNF-α和部分IL-1β的表达。本研究表明,NRICM101在牙周炎细胞和动物模型中均具有保护性抗炎作用。抑制HGFs中促炎细胞因子的表达,减轻结扎性牙周炎大鼠牙槽骨丢失。对IL-6和TNF-α的影响是明显的,而IL-1β的部分减少可能反映了其他的调节机制,如炎性体激活,这需要进一步的研究。这些发现表明,NRICM101可以减少炎症而不产生细胞毒性,并支持其在治疗慢性牙周炎症方面的潜在应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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