Use of 1-deoxynojirimycin for preparing diabete and disney disease drug
A technology of deoxynojirimycin, diabetic nephropathy, applied in the field of pharmacy
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Embodiment 1
[0027] Example 1 Effect of 1-deoxynojirimycin on proliferation of rat mesangial cells cultured in high glucose
[0028] 1. Materials and Reagents
[0029] The rat mesangial cell line was donated by Professor Mei Changlin, Department of Nephrology, Shanghai Changzheng Hospital Affiliated to Second Military Medical University. Reagents: 1-deoxynojirimycin is a product of Wako, Japan; MTT and DMSO are products of AMRESCO; Trypan Blue is a product of Sigma, USA. The microplate reader was produced by Labsystem MK3 in Finland.
[0030] 2. Method
[0031] 2.1 The effect of high glucose on the proliferation of mesangial cells
[0032] Experimental grouping: normal glucose group (NG): the concentration of D-glucose is 5 mmol / L. High glucose group (HG): D-glucose concentration is 30mmol / L. Each group has 6 replicate wells. Hypertonic group (mannitol group): D-glucose (5mmol / L)+mannitol (25mmol / L).
[0033] Experimental method: Cells were inoculated in 96-well cell culture plates ...
Embodiment 21
[0060] Example 21 - Effect of deoxynojirimycin on the expression of α-SMA in rat mesangial cells cultured in high glucose
[0061] 1. Materials and reagents:
[0062] α-smooth muscle actin Wuhan Boster Biological Company, diluted 1:200 with sterile water when used. Rat type IV collagen (type IV collagen), rat fibronectin (Fibronectin, FN) ELISA kits were purchased from Shanghai Senxiong Biological Co., Ltd. UltraSensitive TM S-P immunohistochemical staining kit, diaminobenzidine (DAB) chromogen, Fuzhou Maixin Biotechnology Co., Ltd.
[0063] 2. Method
[0064] 2.1 Experimental grouping and drug intervention training
[0065] Experimental grouping: ① normal glucose group (NG): 5mmol / L D-glucose; ② high glucose group (HG): 30mmol / L D-glucose; ③ mannitol group (M): 5mmol / L D-glucose + 25mmol / L L mannitol; ④ DNJ group: D-glucose 30mmol / L + DNJ 5mmol / L, and a normal glucose negative control group (without primary antibody).
[0066] Drug intervention time: Immunocytochemical...
Embodiment 3
[0075] Example 3 1-deoxynojirimycin on the synthesis of type IV collagen and fibers in rat mesangial cells cultured in high glucose
[0076] The effect of connexin
[0077] 1. Materials and Reagents
[0078] Rat type IV collagen typeIVcollagen, rat fibronectin (Fibronectin) ELISA kits were purchased from Shanghai Senxiong Technology Industrial Co., Ltd.
[0079] 2. Experimental method
[0080] 2.1 ELISA method to determine the expression of type IV collagen and fibronectin
[0081] MSc cells were inoculated in 24-well plates according to conventional methods, and after being synchronized in the G0 phase, they were divided into normal glucose concentration (NG) group: containing D-glucose 5mmol / L; high glucose concentration (HG) group: containing D-glucose 30mmol / L L; Mannitol group (M) group: D-glucose 5mmol / L+25mmol / L mannitol; DNJ group: D-glucose 30mmol / L+1-deoxynojirimycin (DNJ). After 24h, 48h, and 72h of intervening cells, the supernatant was col...
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