Li Zhong, Cheng Jun-Min, Peng Hong-Xing, Jiang Xing-Yu, Gong Jian, Xiao Min
Jiangxi Provincial Hospital of Integrated Chinese and Western Medicine Nanchang 330000, China the Fourth Affiliated Hospital of Jiangxi University of Chinese Medicine Nanchang 330000, China.
Hubei Provincial Hospital of Traditional Chinese Medicine Wuhan 430074, China.
Zhongguo Zhong Yao Za Zhi. 2022 Nov;47(22):6191-6198. doi: 10.19540/j.cnki.cjcmm.20220613.706.
This study aims to explore the effect of Sini Decoction on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB) signaling pathway in the mice with allergic asthma(AA). Forty-eight SPF-grade BALB/c mice were randomly assigned into a blank control group, a model group, a dexamethasone group, and high-, medium-, and low-dose Sini Decoction groups, with 8 mice in each group. The sensitization solution made of ovalbumin and aluminum hydroxide powder was injected intraperitoneally in other groups except the blank control group which was injected with an equal volume of normal saline. The solution(or normal saline) was injected three times in total with an interval of 7 days. At the same time of sensitization, external cold stimulation and ice water were administered in a 4 ℃ climate box for 20 min every day. After modeling, the mice in each group were administrated with corresponding drugs by gavage for 3 weeks. At the end of administration, pentobarbital sodium(30 mg·kg~(-1)) was used for anesthesia, and then the samples were collected for the determination of various indexes. The phenol red test was conducted to evaluate tracheal excretion function. The histopathological changes of lung tissue were observed via hematoxylin-eosin(HE) staining. Masson staining was employed to reveal the deposition of blue collagen fibers around bronchi in lung tissue and the area occupied by blue collagen fibers was calculated. Immunofluorescence method was used to measure the expression of bronchial type Ⅰ collagen(Col-Ⅰ) and α-smooth muscle actin(α-SMA). The protein and mRNA levels of TLR4, NF-κB, cysteinyl aspartate specific proteinase-1(caspase-1), and interleukin-13(IL-13) were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction(real-time PCR), respectively. Compared with the model group, Sini Decoction significantly increased the phenol red excretion from trachea, lowered the lung inflammation score, reduced subepithelial collagen deposition, and decreased Col-Ⅰ and α-SMA levels. Furthermore, the decoction down-regulated the protein and mRNA levels of TLR4, NF-κB, caspase-1, and IL-13 in mouse lung tissue. In conclusion, Sini Decoction can improve air remodeling by inhibiting the TLR4/NF-κB signaling pathway.
本研究旨在探讨四逆汤对过敏性哮喘(AA)小鼠Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路的影响。将48只SPF级BALB/c小鼠随机分为空白对照组、模型组、地塞米松组、四逆汤高、中、低剂量组,每组8只。除空白对照组腹腔注射等体积生理盐水外,其他组腹腔注射由卵清蛋白和氢氧化铝粉末制成的致敏液。共注射3次,间隔7天。致敏同时,将小鼠置于4℃气候箱中每天给予外部冷刺激和冰水20分钟。造模后,各组小鼠灌胃给予相应药物3周。给药结束后,用戊巴比妥钠(30 mg·kg⁻¹)麻醉小鼠,然后采集样本测定各项指标。进行酚红试验以评估气管排泄功能。通过苏木精-伊红(HE)染色观察肺组织的组织病理学变化。采用Masson染色显示肺组织支气管周围蓝色胶原纤维的沉积情况,并计算蓝色胶原纤维所占面积。采用免疫荧光法检测支气管Ⅰ型胶原(Col-Ⅰ)和α-平滑肌肌动蛋白(α-SMA)的表达。分别通过蛋白质免疫印迹法和实时荧光定量聚合酶链反应(实时PCR)测定TLR4、NF-κB、半胱天冬酶-1(caspase-1)和白细胞介素-13(IL-13)的蛋白和mRNA水平。与模型组相比,四逆汤显著增加气管酚红排泄量,降低肺部炎症评分,减少上皮下胶原沉积,并降低Col-Ⅰ和α-SMA水平。此外,四逆汤下调小鼠肺组织中TLR4、NF-κB、caspase-1和IL-13的蛋白和mRNA水平。综上所述,四逆汤可通过抑制TLR4/NF-κB信号通路改善气道重塑。