Effect of Ganlu Xiaodu Micropills and its decoction parts on the expression of inflammatory factors in NR8383 cells treated with Mycoplasma pneumoniae
ZHENG Hao1 ZHANG Baoqing2▲ ZHOU Xu2
1.The First Clinical Medical School, Shandong University of Traditional Chinese Medicine, Shandong Province, Jinan 250355, China;
2.Department of Pediatrics of Traditional Chinese Medicine, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Shandong Province, Jinan 250014, China
Abstract：Objective To study the effect of Ganlu Xiaodu Micropills and its decoction on the expression of inflammatory factors in rat alveolar macrophages (NR8383) infected with Mycoplasma pneumoniae (MP). Methods Thirty-five female Wistar rats of SPF grade aged eight weeks, weighing (200±10) g, were selected. According to the random number table method, they were divided into blank serum group (normal saline 2 ml/d), Ganlu Xiaodu Micropills serum group (33 g/[kg·d]), Azithromycin serum group (0.063 mg/[kg·d]), decoction part 1 serum group (12.3 g/[kg·d]), decoction part 2 serum group (20.8 g/[kg·d]), with seven rats in each group. Each group of doses was given once a day for one week to prepare drug- containing serum. NR8383 cells were treated with blank serum and drug-containing serum, respectively, CCK-8 assay was used to detect and compare the difference of viability between NR8383 cells treated with different serum and untreated cells. NR8383 cells were divided into control group, model group, Ganlu Xiaodu Micropills group, Azithromycin group, and decoction part 1, 2 groups. Except for the control group, the other groups were treated with 106-107 CCU/ml MP bacterial solution for four hours to establish MP infection model, and then the corresponding drug-containing serum was added and incubated for 24 hours. The cells and supernatant were collected, and qPCR and Western blot were used to detect the mRNA and protein expression of Toll-like receptor 4 (TLR4), MyD88, and nuclear factor κB (NF-κB) in each group. The expression levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-10 in each group were detected by enzyme-linked immunosorbent assay. Results The survival rate of cells treated with each serum were lower than those of cells without treatment (P＜0.05). There was no significant difference in the survival rate of cells treated with different serum (P＞0.05). The expression levels of TLR4, MyD88, and NF-κB-p50 mRNA and protein in the model group were higher than those in the control group, and the expression levels of TLR4, MyD88, and NF-κB-p50 mRNA and protein in the Ganlu Xiaodu Micropills group and the decoction part 2 group were lower than those in the model group, the expression of MyD88 mRNA and TLR4 protein in the decoction part were lower than those in the model group (P＜0.05). The mRNA expression of TLR4 and MyD88, and the protein levels of TLR4, MyD88, NF-κB-p50 in Ganlu Xiaodu Micropills group were lower than those in decoction part 1 group, and the mRNA expression of NF-κB-p50 and the protein level of TLR4 were higher than those in decoction part 2 group (P＜0.05). The levels of TNF-α and IL-1β in Ganlu Xiaodu Micropills group and decoction part 2 group were lower than those in model group, the level of IL-1β in decoction part 1 group was lower than that in model group, and the level of IL-10 in decoction part 2 group was lower than that in model group (P＜0.05). The levels of TNF-α, IL-1β, and IL-10 in Ganlu Xiaodu Micropills group were lower than those in decoction part 1 group, and the levels of TNF-α and IL-1β were higher than those in decoction part 2 group (P＜0.05). Conclusion MP infection can promote the expression of TNF-α, IL-1β, and IL-10 in NR8383 cells. Ganlu Xiaodu Micropills and its decoction containing serum can reduce the inflammatory response by inhibiting the expression of TNF-α and IL-1β, and the mechanism may be related to the TLR4/MyD88/NF-κB pathway.
郑豪1 张葆青2▲ 周旭2. 甘露消毒丹及其拆方对肺炎支原体干预的NR8383细胞炎症因子表达的影响[J]. 中国医药导报, 2023, 20(18): 4-9.
ZHENG Hao1 ZHANG Baoqing2▲ ZHOU Xu2. Effect of Ganlu Xiaodu Micropills and its decoction parts on the expression of inflammatory factors in NR8383 cells treated with Mycoplasma pneumoniae. 中国医药导报, 2023, 20(18): 4-9.
京公网安备 11010502046598号