| Euryale ferox Salisb. is a mature seed of Euryale genus of Nymphaeaceae. It isorigin produced in East Asia and India. It is mainly produced in Henan, Ahhui,Hubei and other places in China. It is used as medicinal plants of treatment for somechronic diseases, which has a high pharmacological value. The outer package ofeuryale ferox seeds is a hard shell. In the processing of food industrial production,the euryale ferox shells (ES) are a large number of abandoned, which caused a wasteof shells resources. The hypoglycemic and molecular mechanisms of triterpenoid-rich extracts from ES is analyzed in streptozotocin-induced (STZ) diabetic mice,which is provided the experimental basis for development and utilization of ES.The broken shells were ground into fine powder and soaked in70%ethanol at65C for16h for the extraction. The concentrated extract was then diluted withwater, and the solution was extracted3times with petroleum ether, ethyl acetate andwater-saturated butanol, in that order. To determine the content of triterpenoid in ESby Spectrophotometry and investigate the constituents of triterpenoid in extracts byhigh performance liquid chromatography-mass spectrometry (LC-MS). Body weight,organ weight index and lipid profile parameters were observed in normal anddiabetic mice. The hypoglycemic activity was assessed by fasting blood glucose(FBG) and fasting insulin (FINS) to calculate the insulin sensitivity index (ISI). ThemRNA expression of Fox01, PI-3K, PKB, GLUT2were examined by RT-PCR andprotein level in the liver were determined using enzyme linked immunosorbent assay.In addition, the potentially regulative mechanisms on insulin resistance (IR) werediscussed.The results showed that the total mass fractions triterpenoid in ES extracts was36.7%, which was determined using5%vanillin-acetic acid and perchloric acid ascolor system. LC-MS in positive mode was used to get the mass spectrum of theanalyte. The molecular mass of the main component was at about325and512, predicted molecular formula may be C23H18O13and C25H20O12. The extracts of EScould inhibit reduction in the body weight of diabetic mice and regulate glucosemetabolism, which suggested that the ES extracts could effectively reverse theabnormal enlargement of the liver and spleen. The ISI and C-P levels in thehigh-dose feeding group was significantly higher than that in the model group(p<0.05), with both nearing the normal level. The hypolipidemic action after thisextract supplementation was confirmed by significant (p<0.05) decreases in thelevels of total cholesterol (T-CHO), low density lipoprotein (LDL-C) andtriglycerides (TG) and increase in low-density lipoprotein (HDL-C) compared withthe untreated diabetic mice (p <0.01). These included that the ES extracts reducedthe mRNA expression of Fox01, and raised the gene expression of PI-3K and PKB.Additionally, an significantly increase in the phosphorylation of the PI-3K and PKBprotein level was observed (P<0.05), which also reduced the protein expression ofFox01.In summary, the study of the hydroalcoholic extracts of ES by LC–MSsuggested the presence of triterpenoids. In STZ-induced type2diabetes mice, thetriterpenoid-rich extracts from the ES could inhibit organ enlargement caused bydiabetes and restored the essential function of the liver and kidney, changes in thelipid profile play an important role in converting glucose into glycogen, it alsoprovided a protective effect to the diabetic mice similar to normal and enhanced theISI. The ES extracts possibly acts to restore the intracellular energy balanceconfirmed by inhibition of Fox01, regulating glucose uptake by way of PI-3K andPKB pathway activation, which screened molecular targets about protective effectson beta cells and improving function of ES extracts on insulin resistance (IR).Moreover, we confirmed that ES extracts regulated the expression level oftranscription and translation about IR. |
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