RESUMO
BACKGROUND: Diabetes occurs due to insulin deficiency or less insulin. To manage this condition, insulin administration as well as increased insulin sensitivity is required, but exogeneous insulin cannot replace the sensitive and gentle regulation of blood glucose levels same as ß cells of healthy individuals. By considering the ability of regeneration and differentiation of stem cells, the current study planned to evaluate the effect of metformin preconditioned buccal fat pad (BFP) derived mesenchymal stem cells (MSCs) on streptozotocin (STZ) induced diabetes mellitus in Wistar rats. MATERIALS & METHODS: The disease condition was established by using a diabetes-inducing agent STZ in Wistar rats. Then, the animals were grouped into disease control, blank, and test groups. Only the test group received the metformin-preconditioned cells. The total study period for this experiment was 33 days. During this period, the animals were monitored for blood glucose level, body weight, and food-water intake twice a week. At the end of 33 days, the biochemical estimations for serum insulin level and pancreatic insulin level were performed. Also, histopathology of the pancreas, liver and skeletal muscle was performed. RESULTS: The test groups showed a decline in the blood glucose level and an increase in the serum pancreatic insulin level as compared to the disease group. No significant change in food and water intake was observed within the three groups, while body weight was significantly reduced in the test group when compared with the blank group, but the life span was increased when compared with the disease group. CONCLUSION: In the present study, we concluded that metformin preconditioned buccal fat pad-derived mesenchymal stem cells have the ability to regenerate damaged pancreatic ß cells and have antidiabetic activity, and this therapy is a better choice for future research.
Assuntos
Diabetes Mellitus Experimental , Células-Tronco Mesenquimais , Metformina , Ratos , Animais , Metformina/farmacologia , Metformina/uso terapêutico , Ratos Wistar , Glicemia , Células-Tronco Mesenquimais/patologia , Insulina/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/patologia , Peso CorporalRESUMO
The present study is the first report of utilizing Tithonia rotundifolia weed as a substrate for inulinase production from Fusarium solani JALPK. It also deals with the statistical optimization of culture conditions to enhance the enzyme yield. Amongst the 11 variables screened by Plackett- Burman design, Inulin in combination with Agave sisalana extract, Tithonia rotundifolia extract and NaNO3 had a significant influence on inulinase production and their concentrations were further optimized employing Box Behnken design. An enhancement of inulinase production from 970â¯EU/mL to 3261.011â¯EU/mL was gained after media optimization. Amongst the screened carbon sources Tithonia rotundifolia was found to be very effective in stimulating elevated inulinase synthesis. The Tithonia rotundifolia weed extract was treated with inulinase from Fusarium solani JALPK to form fructose which was estimated spectrophotometrically. This liberated fructose was also confirmed by osazone formation test and FTIR. HPTLC analysis of product revealed the exoinulinase nature of the enzyme produced by Fusarium solani JALPK since fructose was the only end product after hydrolysis of inulin rich weed in fermented broth. Thus the elevated extracellular inulinase yielding novel property of Fusarium solani JALPK (KY914560) contributes in considering it as a potential candidate with food, pharmaceutical and bioremediation applications.
Assuntos
Proteínas Fúngicas/metabolismo , Fusarium/enzimologia , Glicosídeo Hidrolases/metabolismo , Extratos Vegetais/química , Plantas Daninhas/química , Agave/química , Agave/microbiologia , Meios de Cultura/química , Meios de Cultura/metabolismo , Fermentação , Frutose/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Fusarium/química , Fusarium/genética , Fusarium/metabolismo , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/genética , Hidrólise , Inulina/química , Inulina/metabolismo , Extratos Vegetais/metabolismo , Plantas Daninhas/microbiologiaRESUMO
Tithonia rotundifolia is an easily available and abundant inulin rich weed reported to be competitive and allelopathic. This weed inulin is hydrolyzed by inulinase into fructose. Response surface methodology was employed to optimize culture conditions for the inulinase production from Arthrobacter mysorens strain no.1 isolated from rhizospheric area of Tithonia weed. Initially, Plackett- Burman design was used for screening 11 nutritional parameters for inulinase production including inulin containing weeds as cost effective substrate. The experiment shows that amongst the 11 parameters studied, K2HPO4, Inulin, Agave sisalana extract and Tithonia rotundifolia were the most significant variables for inulinase production. Quantitative effects of these 4 factors were further investigated using Box Behnken design. The medium having 0.27% K2HPO4, 2.54% Inulin, 6.57% Agave sisalana extract and 7.27% Tithonia rotundifolia extract were found to be optimum for maximum inulinase production. The optimization strategies used showed 2.12 fold increase in inulinase yield (1669.45 EU/ml) compared to non-optimized medium (787 EU/ml). Fructose produced by the action of inulinase was further confirmed by spectrophotometer, osazone, HPTLC and FTIR methods. Thus Tithonia rotundifolia can be used as an eco-friendly, economically feasible and promising alternative substrate for commercial inulinase production yielding fructose from Arthrobacter mysorens strain no.1.