Greetings first
I am working with a model of diabetes 2 using Streptozotocin in citrate buffer (4.5) and Nicotinamide in saline (0.9), in the proportions of 65 mg / kg and 225 mg / kg respectively, both intraperitoneally and leaving 15 minutes of at each other,. However, I had serious problems at the time of the induction. In my first attempt I induced 42 rats with the described method and 5 days later I evaluated the fasting glucose (12 h) and all the rats were in the ranges of normoglycemic. In this case I attributed the problem to the time I had the STZ in solution (greater than 20 min). However in my second attempt (6 rats) I used STZ freshly diluted in fresh buffer and also covered the container with aluminum foil and my results were:
r1: 140 mg / dl of glucose in blood
r2: 129
r3: 515
r4: 95
r5: 141
r6: 496
It should be noted that for the measurement I used an Accucheck Active brand blood glucose meter and the blood was obtained from the tip of the tail. In addition, the measurement was made 3 days after the induction and 120 minutes after having eaten food.
I have several doubts about these results,:
1- Rats 3 and 6 have glucose levels that are considered diabetic, but I do not know if they are too high for a type 2 model and maybe have obtained a type 1 model.
2-According to the review of Asghar Ghasemi, 2014 most reports use parameters within ≥ 250-140 to consider diabetics to rats, in this case could consider rats 1,3,5,6 diabetics, although their values are so different?
3- The 120 minutes of fasting that I used to make the measurement are considered non-fasted or in any case what exactly does that term mean?
4- What would be the best fasting period to be able to perform the glucose measurement?
I hope you can help me with my problem, because this project is my final undergraduate project.
Thank you very much in advance
Victor, there is no issue in your protocol, all is good. But check the glucose level of same animals after 10-12 days (approx time to reach steady hyperglycemic state ). I think you will get hyperglycemia in animals above 250 mg/dl for sure.
Victor, there is no issue in your protocol, all is good. But check the glucose level of same animals after 10-12 days (approx time to reach steady hyperglycemic state ). I think you will get hyperglycemia in animals above 250 mg/dl for sure.
Dear Dr. Victor
I agree with Dr. Kuldeep answers
There is no problem in your protocol procedures. Measure the glucose levels of same animals after at least 10-12 days (approx time to reach steady hyperglycemic state )..
Best regards
Dear Dr. Victor,
1. What is the age of rats used in the experiment? I believe older rats are not suitable for experiments. You might want to see the following paper:
Rat’s age versus human’s age: What is the relationship? March 2012 Arquivos brasileiros de cirurgia digestiva : ABCD = Brazilian archives of digestive surgery 25(1):49-51DOI: 10.1590/S0102-67202012000100011
http://www.scielo.br/pdf/abcd/v25n1/en_11.pdf
2. Did you try the neo-natal rats for inducing diabetes with STZ?
Dear Victor,
Try to use distilled water for disolving stz and give a dose of 55mg/kg. We used this and succeeded.
thank you all for your answers, they have helped me a lot
:)
Hello Victor Marcelo Utrilla , how r u? I hope great!
Hello Victor Marcelo Utrilla , how r u? I hope great!
I worked with experimental diabetes models and I was really terrible when the induction doesn´t occurs!! Sometimes it could be caused for a little missing step that you can consert! So, let me help you! I worked with type 1 diabetic model and used Alloxan for this porpoise!
The question is: Which type of diabetes would you like to create? Which interquartile glycemic measurement to follow? It could be severe (>500mg/dl), moderate (250-400mg/dl) or mild diabetes (150-250mg/dl). Ok, that’s the first point to consider when you choose your concentration and route of administration!!!!
Helping in your questions:
1- Rats 3 and 6 have glucose levels that are considered diabetic, but I do not know if they are too high for a type 2 model and maybe have obtained a type 1 model. – you’ll have to observe them very closely!! How are they clinically? Does the insulin necessary? When they became insulin dependent, I mean, with symptons of DKA and couldn’t survive without treatment – seems to be a type 1 behaviour!
2-According to the review of Asghar Ghasemi, 2014 most reports use parameters within ≥ 250-140 to consider diabetics to rats, in this case could consider rats 1,3,5,6 diabetics, although their values are so different? – supposedly, yes! Will you have to cheking their glucose concentrations daily and accompanying the symptons (behavior, ingestion of food and water, weight) – the mild models can recover the hyperglycemic state very fast!
3- The 120 minutes of fasting that I used to make the measurement are considered non-fasted or in any case what exactly does that term mean? This time it’s good to cheking the results of your induction – the question is: Do are they submitted to fasting period before the injection? Some researchers suggest 12-14 hours fasting before the experiment and It’s a point considering determinant to your success!!!! If they doesn’t had this previous fasting period, they coudn’t became diabetic!
4- What would be the best fasting period to be able to perform the glucose measurement? If these previous recommendations would complete, in 48 hours you’ll have your diabetic models!!! I recommend to check each 3 days to attend their status!
I hope that I could helped!
Don't hesitate in contact if it's necessary!!
Good look!
Best regards!!
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