Nan Yang1, Zibo Dong2, Gang Tian2, Maomao Zhu3, Chao Li4, Weiquan Bu5, Juan Chen6, Xuefeng Hou6, Ying Liu6, Gang Wang6, Xiaobin Jia6, Liuqing Di3, Liang Feng7. 1. Post-doctoral Research Center, Nanjing University of Chinese Medicine & Jumpcan Pharmaceuticl Co., Ltd, Jiangsu, Taixing 225441, PR China; School of Pharmacy, Nanjing University of Chinese Medicine, 100# Shizi Road, Nanjing, Jiangsu 210023, PR China; Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, Jiangsu 210028, PR China. 2. Post-doctoral Research Center, Nanjing University of Chinese Medicine & Jumpcan Pharmaceuticl Co., Ltd, Jiangsu, Taixing 225441, PR China; Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, Jiangsu 210028, PR China. 3. School of Pharmacy, Nanjing University of Chinese Medicine, 100# Shizi Road, Nanjing, Jiangsu 210023, PR China. 4. Post-doctoral Research Center, Nanjing University of Chinese Medicine & Jumpcan Pharmaceuticl Co., Ltd, Jiangsu, Taixing 225441, PR China. 5. Department of Pediatrics, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, Jiangsu 210028, PR China. 6. School of Pharmacy, Nanjing University of Chinese Medicine, 100# Shizi Road, Nanjing, Jiangsu 210023, PR China; Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, Jiangsu 210028, PR China. 7. Post-doctoral Research Center, Nanjing University of Chinese Medicine & Jumpcan Pharmaceuticl Co., Ltd, Jiangsu, Taixing 225441, PR China; School of Pharmacy, Nanjing University of Chinese Medicine, 100# Shizi Road, Nanjing, Jiangsu 210023, PR China; Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, Jiangsu 210028, PR China. Electronic address: wenmoxiushi@163.com.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: TMHM is a type of Chinese medicine commonly used in medical practice and has multiple functions, including clearing heat, detoxification, reducing swelling, and tumor therapy. Previous research has demonstrated that the OAC of TMHM (TMHM-OAC) displays advantageous therapeutic action against respiratory inflammation. However, the effect of TMHM-OAC on inflammatory injury and its anti-inflammatory role requires further clarification. MATERIALS AND METHODS: An in vitro inflammation damage model was employed using NHBE cells and 100ng/ml of (LPS). HPLC-DAD was conducted to analyze the components of TMHM-OAC. An ELISA was conducted to determine IL-1β, IL-6, TNF-α, and NO expression. An MTT assay was conducted to determine the cytotoxicity of TMHM-OAC. The levels of IL-1β, IL-6, TNF-α, caspase-3, caspase-8, iNOS, TLR4p-nuclear factor kappa-B kinase (p-IκκB), and p-NF-κB p65 in cellular protein, as well as the mRNA levels, were determined using WB, IF testing, and Q-PCR. RESULTS: TMHM-OAC significantly reduced LPS-induced NHBE cell inflammation, which was reflected in the reduced expression of relevant cytokines such as TNF-α, IL-1β, IL-6 and NO, caspase-3, and caspase-8. In addition, this component suppressed TLR4, p-IKKβ, and p-NF-κB p65 levels in both mRNA and cellular protein. CONCLUSION: TMHM-OAC can reduce LPS-induced inflammation in NHBE cells and this function could be linked to the regulation of the TLR4/IKK/NF-kB pathway.
ETHNOPHARMACOLOGICAL RELEVANCE: TMHM is a type of Chinese medicine commonly used in medical practice and has multiple functions, including clearing heat, detoxification, reducing swelling, and tumor therapy. Previous research has demonstrated that the OAC of TMHM (TMHM-OAC) displays advantageous therapeutic action against respiratory inflammation. However, the effect of TMHM-OAC on inflammatory injury and its anti-inflammatory role requires further clarification. MATERIALS AND METHODS: An in vitro inflammation damage model was employed using NHBE cells and 100ng/ml of (LPS). HPLC-DAD was conducted to analyze the components of TMHM-OAC. An ELISA was conducted to determine IL-1β, IL-6, TNF-α, and NO expression. An MTT assay was conducted to determine the cytotoxicity of TMHM-OAC. The levels of IL-1β, IL-6, TNF-α, caspase-3, caspase-8, iNOS, TLR4p-nuclear factor kappa-B kinase (p-IκκB), and p-NF-κB p65 in cellular protein, as well as the mRNA levels, were determined using WB, IF testing, and Q-PCR. RESULTS:TMHM-OAC significantly reduced LPS-induced NHBE cell inflammation, which was reflected in the reduced expression of relevant cytokines such as TNF-α, IL-1β, IL-6 and NO, caspase-3, and caspase-8. In addition, this component suppressed TLR4, p-IKKβ, and p-NF-κB p65 levels in both mRNA and cellular protein. CONCLUSION:TMHM-OAC can reduce LPS-induced inflammation in NHBE cells and this function could be linked to the regulation of the TLR4/IKK/NF-kB pathway.