The data of study about the chitosan-selenium nanoparticles on depression-like behaviour induced by fluoride in mice via JAK2-STAT3 pathway

Published: 6 August 2021| Version 1 | DOI: 10.17632/brjjkmdb62.1
Contributor:
Jinming Wang

Description

150 mg/L NaF exposure significantly decreased the protein expressions of JAK2 and STAT3 in Raw data 39-40. However, the expressions of p-JAK2 and p-STAT3 were increased in the F group compared to their respective total proteins, indicating that F affected JAK2 and STAT3 and its phosphorylation form, promoting the activation of the JAK2-STAT3 signaling pathway in Raw data 41-42. F and 2 mg/kg·bw CS-SeNPs both enhanced the protein expression levels of IL-1β (1.623 times, 2.089 times) and IL-6 (2.05 times, 4.556 times) whereas 1 mg/kg·bw CS-SeNPs diminished the IL-1β secretion induced by F in Raw data 44-45. 1 mg/kg·bw CS-SeNPs diminished F-enhanced phosphorylation of STAT3, significantly reduced F-induced IL-1β as well as IL-6 production, indicating that 1 mg/kg·bw CS-SeNPs alleviated cortical inflammatory impairment triggered by F through the JAK2/STAT3 signaling pathway. These data provide credible data support for related pathway studies for comparison or analysis.

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The data can be obtained as follows: Raw data 34-38:Total RNA from the 50 mg cortex was extracted using 1 mL Trizol reagent, and was reversely transcribed at 37 ℃ for 15 min, 85 ℃ for 5 s and 4 ℃ for 10 min with the PrimeScript™ kit (Takara, Dalian, China). QRT-PCR was conducted using SYBR Premix Ex Taq™ II (Takara, Dalian, China) on the Mx3000P™ QPCR system (Stratagene, California, USA). The reaction conditions are: (1) predegeneration raction 1 cycle: 95 ℃ for 30 s for, (2) Polymerase chain reaction 40 cycle: 95℃ for 5s, 60℃ for 30s, 72℃ for 30s, (3) Dissociation protocol 1 cycle: 95℃ for 15s, 60℃ for 1 min, 95℃ for 15s. The CDS sequences were retrieved from the NCBI and primer were designed by Primer 3 plus, according to their mRNA sequences of genes (Table 2). The relative mRNA expression of the target genes was calculated using the 2-∆∆CT method to β-actin, a housekeeping gene. (1)△CT (control group) =CT (target gene) -CT (internal reference gene) (2)△CT (experimental group) =CT (target gene) -CT (internal reference gene) (3) Calculate the mean value of △CT in the control group (4)△△CT=△CT (control group and experimental group) - the mean value of control group △CT (5)2-△△CT value was the relative expression level of the target gene. The raw data were analysed by the GraphPad Prism 6.01 software. Raw data 39-46:To get the total protein, 40 mg of cortex tissue was weighed and placed into 396 mL RIPA lysis buffer at 4 ℃ for homogenization containing 4 mL PMSF and 4 mL phosphatase inhibitor liquid A and B. 70 μg total protein was added to 10% SDS polyacrylamide electrophoresis gels. After performing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the electrophoretically separated components of JAK2, STAT3, p-JAK2 and p-STAT3 were transferred from the gel to the NC membrane at 300 mA for 30 min, and other target proteins were transferred to the NC membrane at 35 V for 70 min. The NC membrane was placed face-up in 5% skim milk powder for blocking and incubated at 4 ℃overnight with a primary antibody diluted to the appropriate concentration, namely rabbit anti-STAT3, p-STAT3, p-JAK2 polyclonal antibody (1 : 500, ABclonal, Wuhan, China), rabbit anti-JAK2 monoclonal antibody (1 : 500, ABclonal, Wuhan, China), rabbit anti-β-actin, IDO1 polyclonal antibody (1:1000, proteintech, Wuhan, China), rabbit anti-IL-6, TNF-α, IL-1β polyclonal antibody (1:500, Bioss, Beijing, China). The ECL reagent was evenly coated on the NC membrane for 2 min, and images were collected, and the optical density of the target band was analyzed using a Fluor Chem Q System (Alpha Innotech, CA, USA). The raw data were analysed by the GraphPad Prism 6.01 software.