Application of sglt2 inhibitors in regulating inflammation
An inhibitor and inflammation technology, applied in the application field of sodium-glucose co-transporter 2 inhibitors in the regulation of inflammation, can solve the problems of poor patient compliance and lack of anti-inflammatory drugs, and achieve the effect of improving inflammatory symptoms
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Embodiment 1
[0046] Example 1. In vitro lipopolysaccharide-induced inflammatory cell model creation and activity evaluation
[0047] 1. Experimental materials
[0048] 1. Compound
[0049] SGLT2 inhibitors (CAN, Empagliflozin, Dapagliflozin, Bexagliflozin, Ipragliflozin and Tofogliflozin hydrate), Metformin hydrochloride, Rosglitazone, Glibenclamide, Salsalate, Liraglutide, Sitagliptin and Insulin were purchased from MedChemExpress (MCE) in the United States; chloroform was from Tianjin Hongyan Chemical Reagent Factory, dimethyl sulfoxide (DMSO), absolute ethanol and isopropanol were purchased from Sangon Bioengineering (Shanghai) Co., Ltd.; lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (Lot.No. L4391), dissolved in ultrapure water and stored at -20°C.
[0050] 2. Cell source
[0051] Mouse monocytes / macrophages (RAW264.7) and human peripheral blood mononuclear cells (THP-1) were purchased from the Cell Resource Center, Shanghai Institutes of Biological Sciences, Chinese Aca...
Embodiment 2
[0103] Example 2, Research on anti-inflammatory mechanism of SGLT2 inhibitor represented by CAN
[0104] 1. Experimental materials
[0105] 1. Biochemical reagents
[0106] p62 (PM045) was from Medical & Biological Laboratories; LC3 (L7543) was from Sigma-Aldrich; β-actin antibody (A1978) was from Sigma-Aldrich; the glucose kit was purchased from Zhongsheng Beikong Biotechnology Co., Ltd.; Intracellular glucose metabolism test kit (Agilent Seahorse XFpGlycolysis Stress Test Kit, XFp FluxPak, 103022-100, American Agilent Technologies, Inc. company); ECL reagent (Pierce TM ECL Western blotting substrate) was purchased from Thermo Fisher Scientific, USA).
[0107] 2. Primer sequence
[0108] p62 (Forward: 5'-GAACTCGCTATAAGTGCAGTGT-3'; Reverse: 5'-AGAGAAGCTATCAGAGAGGTGG-3') was purchased from ThermoFisher Scientific (Invitrogen TM )synthesis. The Actin primers in Table 1 of Example 1 were used as controls.
[0109] 3. Instrument
[0110] Fluorescence confocal microscope (FV1...
Embodiment 3
[0157] Example 3: Production of LPS-induced pulmonary inflammation model and activity evaluation in mice
[0158] 1. Experimental materials
[0159] 1. Experimental animals and food
[0160]NIH male mice (about 20 g) were purchased from Guangdong Medical Experimental Animal Center (SPF grade, license number: Yue Jian Zheng Zi, SCXK (Guangdong) 2013-0002). SPF grade food was purchased from Guangdong Medical Experimental Animal Center.
[0161] 2. Chemical reagents
[0162] 4% paraformaldehyde: 4g of paraformaldehyde, prepared with 100ml of normal saline. Anesthetic: 10% urethane injection: 10g urethane, prepared with 100ml normal saline.
[0163] 2. Experimental method
[0164] After the animals in the experimental group and the control group were divided into balanced groups according to body weight, the experiment was started. Firstly, appropriate amounts of LPS and CAN were dissolved in PBS to prepare relevant solutions. 12 hours in advance and 1 hour in advance, the ...
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