2. Methods
2.1. Animals and procedures: 4-5 weeks old male Kunming mice weighing 19-21g were provided by Experimental Animal Centre of Kunming prior to treatment with LOS(10,20mg) or MET(20,40mg), the mice were either treated with vehicle (n=16) or ISO (n=80) for 2 weeks. The mice were fed food and had free access to tap water. The mice were maintained under stable room temperature and a regular 12h dark and light rhythm. Usage of experimental animal was in accordance with the compliance of the National Guidelines for the care and use of Laboratory animals and the guidelines of China Pharmaceutical University approved by licensed number SYXK (Su) 2016-0011 according to the board members of China Pharmaceutical University [29].
2.2. Materials and reagents: DL-Isoproterenol hydrochloride (15627-5G) were brought from Sigma. Metoprolol (Betaloc Zok 50mg) 47.5 mg were brought from AstraZeneca. Losartan Potassium tablets were brought from Nanjing Biological technology Co. Ltd. Other reagents used were Normal Saline and Distilled water to dissolve the tablets [30]. 2.3. Surgical procedure and drug administration: Firstly, 19-21g male mice was taken. These mice were divided into 6 different groups. Firstly, ISO was given via intraperitoneal injection according to the average weight of mice for 2 weeks [30]. After 2 weeks, all the mice were weighed and the …show more content…
Heart tissue was taken out from the Refrigerator and 0.1gm was weighed. Total RNA was extracted using TRIZON Reagent according to the instructions and quantitated by Micro-spectrophotometer. RNA having a ratio of A260/280 1.9-2.0 was reversely transcribed by HiFi-MMLV cDNA kit. Quantitative real-time polymerase chain reaction was performed on a real-time fluorescent quantitative PCR system using ultra SYBR mixture with specific primers[40]. The sequences of primers for mice were as
2.1. Animals and procedures: 4-5 weeks old male Kunming mice weighing 19-21g were provided by Experimental Animal Centre of Kunming prior to treatment with LOS(10,20mg) or MET(20,40mg), the mice were either treated with vehicle (n=16) or ISO (n=80) for 2 weeks. The mice were fed food and had free access to tap water. The mice were maintained under stable room temperature and a regular 12h dark and light rhythm. Usage of experimental animal was in accordance with the compliance of the National Guidelines for the care and use of Laboratory animals and the guidelines of China Pharmaceutical University approved by licensed number SYXK (Su) 2016-0011 according to the board members of China Pharmaceutical University [29].
2.2. Materials and reagents: DL-Isoproterenol hydrochloride (15627-5G) were brought from Sigma. Metoprolol (Betaloc Zok 50mg) 47.5 mg were brought from AstraZeneca. Losartan Potassium tablets were brought from Nanjing Biological technology Co. Ltd. Other reagents used were Normal Saline and Distilled water to dissolve the tablets [30]. 2.3. Surgical procedure and drug administration: Firstly, 19-21g male mice was taken. These mice were divided into 6 different groups. Firstly, ISO was given via intraperitoneal injection according to the average weight of mice for 2 weeks [30]. After 2 weeks, all the mice were weighed and the …show more content…
Heart tissue was taken out from the Refrigerator and 0.1gm was weighed. Total RNA was extracted using TRIZON Reagent according to the instructions and quantitated by Micro-spectrophotometer. RNA having a ratio of A260/280 1.9-2.0 was reversely transcribed by HiFi-MMLV cDNA kit. Quantitative real-time polymerase chain reaction was performed on a real-time fluorescent quantitative PCR system using ultra SYBR mixture with specific primers[40]. The sequences of primers for mice were as