Huang Hefei, He Xuemei, Shi Lingzhu, Yu Jingtao, Lu Zibin, Cao Huihui, Ou Jinying, Chen Xi, Yan Lijun, Yang Jiabin, Zhao Wei, Liu Junshan, Yu Linzhong
Third Level Research Laboratory of State Administration of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, People's Republic of China.
Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, People's Republic of China.
Chin Med. 2024 Feb 14;19(1):24. doi: 10.1186/s13020-024-00893-2.
Encephalitis caused by dengue virus (DENV) is considered a manifestation of severe dengue. Tanreqing injection (TRQ) is a well-known Chinese patented medicine, which has been used to treat brain-related disorders by inhibiting inflammation. Nevertheless, the effects of TRQ on DENV encephalitis have not been studied. The aim of this study was to evaluate the effects of TRQ on DENV encephalitis and to explore its potential mechanisms.
The cytotoxicity of TRQ was examined by MTT assay, and the anti-DENV activities of TRQ in BHK-21 baby hamster kidney fibroblast were evaluated through CCK-8 and plaque assays. The expression levels of NO, IL1B/IL-1β, TNFα and IL6 were measured by qRT‒PCR and ELISA in the BV2 murine microglial cell line. The inhibitory effects of TRQ on NLRP3 inflammasome activation in BV2 cells were examined by Western blotting, qRT‒PCR and ELISA. The effects of TRQ on HT22 mouse hippocampal neuronal cells were examined by CCK-8 assay, morphology observation and flow cytometry. Moreover, a DENV-infected ICR suckling mouse model was developed to investigate the protective role of TRQ in vivo.
TRQ decreased the release of NO, IL6, TNFα and IL1B from BV2 cells and inhibited the activation of NLRP3. The presence of the NLRP3 agonist nigericin reversed the anti-inflammatory activities of TRQ. Furthermore, TRQ inhibited the death of HT22 cells by decreasing IL1B in DENV-infected BV2 cells. In addition, TRQ significantly attenuated weight loss, reduced clinical scores and extended the survival in DENV-infected ICR suckling mice. Critically, TRQ ameliorated pathological changes in ICR suckling mice brain by inhibiting microglia and NLRP3 activation and decreasing the production of inflammatory factors and the number of dead neurons.
TRQ exerts potent inhibitory effects on dengue encephalitis in vitro and in vivo by reducing DENV-2-induced microglial activation and subsequently decreasing the inflammatory response, thereby protecting neurons. These findings demonstrate the potential of TRQ in the treatment of dengue encephalitis.
登革病毒（DENV）引起的脑炎被认为是重症登革热的一种表现形式。痰热清注射液（TRQ）是一种著名的中成药，已被用于通过抑制炎症来治疗脑部相关疾病。然而，TRQ对DENV脑炎的影响尚未得到研究。本研究的目的是评估TRQ对DENV脑炎的影响，并探讨其潜在机制。
通过MTT法检测TRQ的细胞毒性，并通过CCK-8和噬斑试验评估TRQ在BHK-21幼仓鼠肾成纤维细胞中的抗DENV活性。通过qRT-PCR和ELISA在BV2小鼠小胶质细胞系中检测NO、IL1B/IL-1β、TNFα和IL6的表达水平。通过蛋白质免疫印迹法、qRT-PCR和ELISA检测TRQ对BV2细胞中NLRP3炎性小体激活的抑制作用。通过CCK-8试验、形态学观察和流式细胞术检测TRQ对HT22小鼠海马神经元细胞的影响。此外，建立了DENV感染的ICR乳鼠模型，以研究TRQ在体内所起的保护作用。
TRQ减少了BV2细胞中NO、IL6、TNFα和IL1B的释放，并抑制了NLRP3的激活。NLRP3激动剂尼日利亚菌素的存在逆转了TRQ的抗炎活性。此外，TRQ通过降低DENV感染的BV2细胞中的IL1B来抑制HT22细胞的死亡。另外，TRQ显著减轻了DENV感染的ICR乳鼠的体重减轻，降低了临床评分并延长了生存期。至关重要的是，TRQ通过抑制小胶质细胞和NLRP3的激活，减少炎性因子的产生和死亡神经元的数量，改善了ICR乳鼠脑内的病理变化。
TRQ通过减少DENV-2诱导的小胶质细胞激活并随后减少炎症反应，从而在体外和体内对登革热脑炎发挥强大的抑制作用，进而保护神经元。这些发现证明了TRQ在治疗登革热脑炎方面的潜力。
