Objective:To investigate the role and mechanism of PM(2.5)exposure on airway inflammation in juvenile rats based on macrophage recruitment induced by JNK/CCl2 signaling pathway.Methods:A total of 50 juvenile SD rats were randomly divided into 5 groups(n=10).The control group received no treatment,the PM(2.5)group received PM(2.5)particulate matter expo-sure,and the PM(2.5)+Anisomycin group received PM(2.5)exposure and Anisomycin,an activator of JNK,intravenously.Rats in the PM(2.5)+SP600125 group received PM(2.5)exposure and intravenous administration of the JNK inhibitor SP600125,and rats in the PM(2.5)+Pirfenidone group received PM(2.5)exposure and intravenous administration of Pirfenidone,a CCl2 inhibitor.The rats were euthanized and lung tissue was harvested.JNK,phosphorylated JNK(p-JNK)and CCl2 protein expressions were detected by Western blot.Hematoxylin-eosin(HE)staining was used to detect the pathological changes of lung airway tissue and score the pulmo-nary bronchial inflammation.The number of macrophages in alveolar lavage fluid was analyzed by flow cytometry.The levels of IL-6,IL-1β,and TNF-α in alveolar lavage fluid were determined by ELISA.Results:The expression levels of JNK,p-JNK,and CCl2 among all groups(F=205.296,950.408,260.019;all P<0.001)and macrophage content(F=48.414;P<0.001),pulmonary bronchial inflammation score(F=101.703;P<0.001)and IL-6(H=44.890;P<0.001),IL-1β(H=42.071;P<0.001),TNF-α(F=297.154;P<0.001)were statistically significant.Compared with the control group,the expressions of JNK/CCl2 pathway proteins JNK,p-JNK,and CCl2 in PM(2.5)group were significantly up-regulated(all P<0.05),while the content of macrophages was increased(P<0.05),and the pulmonary and bronchial inflammation score was significantly increased(P<0.05).The levels of IL-6,IL-1β,and TNF-α were up-regulated(all P<0.05).Compared with PM(2.5)group,the content of macrophages in PM(2.5)+Anisomycin group was sig-nificantly increased(P<0.05),the pulmonary bronchial inflammation score was significantly increased(P<0.05),and the levels of IL-6,IL-1β,and TNF-α were increased(all P<0.05).The expression levels of JNK,p-JNK,and CCl2 were increased(all P<0.05).Compared with PM(2.5)group,the content of macrophages in PM(2.5)+SP600125 group and PM(2.5)+Pirfenidone group were signifi-cantly decreased(P<0.05),and the pulmonary bronchial inflammation score was significantly decreased(P<0.05).In addition,the levels of IL-6,IL-1β,and TNF-α were significantly decreased(all P<0.05).Compared with PM(2.5)group,the expression levels of JNK,p-JNK,and CCl2 in PM(2.5)+SP600125 group were down-regulated(all P<0.05),and the expression level of CCl2 in PM(2.5)+Pirfenidone group was down-regulated(all P<0.05).Conclusion:JNK/CCl2 pathway induces macrophage recruitment and pro-motes allergic airway inflammation induced by PM(2.5)particulate matter exposure in juvenile rats.

Aim To study the anti-inflammatory activ¬ity of diterpenes from Tripterygium wilfordii on lipopo- lysaccharide ( LPS)-induced macrophage and its mech¬anism. Methods MTT assay was used to detect the cytotoxicity of compounds. The Griess method was used to detect the NO on LPS-induced RAW264. 7 cells. ELISA was applied to determine the contents of inter- leukin 6 (IL-6) , tumor necrosis factor a ( TNF-a ) , interleukin lp (IL-lfj) and interleukin 18 (IL-18) in cell culture supernatant. Western blot was used to de¬tect IkBcx, .INK, ERK, p38, STAT3 and their phos-phorylation in LPS-induced RAW264.7, as well as the effect on COX-2, iNOS, NLRP3, caspase-1 , cleaved- caspase-1. Flow cytometry was employed to detect the effects of compounds on the phagocytosis of RAW 264. 7 cells. Results Hypoglicin II (1) and ent-pimara-8 (14) , 15-diene-19-ol (6) , two diterpenoid compounds from Tripterygium wilfordii could effectively inhibit the expression of inflammatory mediators ( COX-2 and iN- OS) and inflammatory cytokines (IL-6, IL-lp, IL- 18) in LPS-induced RAW264. 7 cells. Further re¬search found that the phosphorylation of IkBcx , JNK, ERK, P38, STAT3 and NLRP3 was all inhibited; however, there was no significant effect on the expres¬sion of IkBcx, JNK, ERK, P38 and STAT3. At the same time, they also inhibited the phagocytosis of mac-rophages. Conclusions The anti-inflammatory mecha¬nism of Tripterygium wilfordii diterpenoids 1 and 6 might be through inhibiting the production of NLRP3 inflammatory bodies, inflammatory mediators (COX-2 and iNOS) and inflammatory cytokines (IL-6, IL-lp and IL-18) , which is closely related to inhibiting the activation of MAPK, NF-kB and STAT3 pathway.

OBJECTIVE: To explore the effect of abnormal miRNA expression on the proliferation of pediatric acute lymphoblastic leukemia (ALL) cells and its related mechanism. METHODS: 15 children with ALL and 15 healthy subjects were collected from the Second Affiliated Hospital of Hainan Medical University from July 2018 to March 2021. MiRNA sequencing was performed on their bone marrow cells, and validated using qRT-PCR. MiR-1294 and miR-1294-inhibitory molecule (miR-1294-inhibitor) were transfected into Nalm-6 cells, and the proliferation of Nalm-6 cells was detected by CCK-8 and colony formation assays. Western blot and ELISA were used to detect apoptosis of Nalm-6 cells. Biological prediction of miR-1294 was performed to find the target gene, which was verified by luciferase reporter assay. Si-SOX15 was transfected into Nalm-6 cells, Western blot was used to detect the expression of Wnt signaling pathway-related proteins and to verify the effect of si-SOX15 on the proliferation and apoptosis of Nalm-6 cells. RESULTS: Compared with healthy subjects, 22 miRNAs were significantly upregulated in bone marrow cells of ALL patients, of which miR-1294 was the most significantly upregulated. In addition, the expression level of SOX15 gene was significantly reduced in bone marrow cells of ALL patients. Compared with the NC group, the miR-1294 group showed increased protein expression levels of Wnt3a and β-catenin, faster cell proliferation, and more colony-forming units, while caspase-3 protein expression level and cell apoptosis were reduced. Compared with the NC group, the miR-1294-inhibitor group showed reduced protein expression levels of Wnt3a and β-catenin, slower cell proliferation, and fewer colony-forming units, while caspase-3 protein expression level was increased and apoptosis rate was elevated. miR-1294 had a complementary base-pair with the 3'UTR region of SOX15 , and miR-1294 directly targeted SOX15 . The expression of miR-1294 was negatively correlated with SOX15 in ALL cells. Compared with the si-NC group, the si-SOX15 group showed increased protein expression levels of Wnt3a and β-catenin, accelerated cell proliferation, and decreased caspase-3 protein expression level and cell apoptosis rate. CONCLUSION MiR-1294 can target and inhibit SOX15 expression, thus activating the Wnt/β-Catenin signaling pathway to promote the proliferation of ALL cells, inhibit cell apoptosis, and ultimately affect the disease progression.
Humans ; Child ; beta Catenin/genetics* ; Wnt Signaling Pathway ; Caspase 3/metabolism* ; Cell Line, Tumor ; MicroRNAs/genetics* ; Cell Proliferation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Apoptosis ; SOX Transcription Factors/metabolism*

Adult ; Humans ; Acute Disease ; Leukemia