Elisa detection kit

First, 50 mg of NP tissues collected from each group was placed in a homogenization tube and added 1 mL of precooled PBS buffer. The tissues were then homogenized and broken in a low-temperature homogenizer. After the completion of homogenization, the homogenization tube was centrifuged at 12,000 g for 10 min at 4 °C to remove cell debris. The supernatant was dispensed into new centrifuge tubes; part of the supernatant was stored at −80 ℃, while the other part was detected for the expression of necrosis factor α (TNF-α), interleukin 6 (IL-6), interleukin 1β (IL-1β), and prostaglandin E2 (PGE2) by using the ELISA detection kit (Nanjing Jiancheng Bioengineering Institute, China).

Serum biochemical detection index kit such as total cholesterol (TC), triglyceride (TG) and ELISA detection kit were purchased from Nanjing Jiancheng Institute of Bioengineering (Jiangsu, China). Blood glucose assay kit was obtained from Jiangsu Yuyue Medical Equipment & Supply Co., Ltd. (Jiangsu, China). All other chemical reagents were analytical grade.

The blood sample was allowed to stand for 30 min and then centrifuged at 4°C at 2000 r/min for 20 min. The supernatant was collected into new centrifuge tubes and stored in a −80°C refrigerator. The serum levels of motilin (MTL), vasoactive intestinal peptide (VIP), leptin, gastrin (GAS), calcitonin gene-related peptide (CGRP), and somatostatin (SS) were detected in accordance with the instructions of the corresponding ELISA detection kit (Jiancheng Bioengineering, Nanjing, Jiangsu, China). The absorbance value at 450 nm was also detected.

Type 2 diabetic mice are known to exhibit specific characteristics, including weight loss, hyperglycemia, and hyperlipidemia. To verify the successful induction of the diabetic model, we measured the levels of body weight, fasting blood glucose, triglyceride, and cholesterol, in our experimental mice. Two weeks after STZ injection, the blood glucose level was determined with a blood glucose meter (Abbott Diabetes Care Ltd., Doncaster, VIC, Australia). The serum levels of triglyceride (TG) and total cholesterol (T-CHO) were measured by commercial detection kits according to the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute, Jiangsu, China).
In addition, the levels of glycated hemoglobin and insulin are considered to be very important detection indicators for diabetes. The concentrations of glycated hemoglobin A1c (HbA1c) and insulin were determined 7 days after the administration of nmFGF1, with a1cNow + meter (PTS Diagnostics, Whitestown, IN, United States) and an enzyme-linked immunosorbent assay (ELISA) detection kit (Nanjing Jiancheng Bioengineering Institute, Jiangsu, China), respectively.

A total of 50 mg of kidney tissue were added to PBS buffer, fully homogenized in a tissue homogenizer, and centrifuged at 4 C, 12000 r/min for 30 min. After that, the supernatant was collected for testing. The level of interleukin (IL)-1β, IL-6, and IL-10 in the kidneys of rats in each group was determined according to the instructions of the corresponding ELISA detection kit (Nanjing Jiancheng, China).