Effect Of Alcohol Extract Of Garcinia Kola Heckel On T2DM Combined With Nafld And Preparation Of Its Granulesabstract

Recently,with the rising incidence of T2 DM combined with NAFLD,the accidence of serious complications have elevated thereupon for threatening people’s health.With the deep development of natural products,many natural products have shown good protective effects on T2 DM combined with NAFLD,and have been widely recognized.As a traditional folk medicine in Africa,Garcinia kola Heckel(GK),modern research on it has shown that it has various pharmacological activities,such as anti-inflammatory,antioxidant,hypolipidemic,and hypoglycemic.However,there are currently no pharmacological studies of GK treatment on T2 DM combined with NAFLD.Therefore,this study aims to explore the therapeutic effect and potential improvement mechanism of ethanol extract of Garcinia kola Heckel(EGK)on T2 DM combined with NAFLD via in vitro and in vivo experiments,and optimize the preparation process of EGK granules to provide a research basis for its subsequent clinical application.In this study,the model of T2 DM combined with NAFLD was established by HGHFD/STZ to study the effect of the EGK on T2 DM combined with NAFLD.After HGHFD treatment for 8 weeks and STZ inducement,the disorder of blood glucose and hyperlipidemia was regarded as the model established successfully.The successfully rats were orally administered with EGK for 8 weeks,and finally euthanized to collect serum and liver tissue.The serum level of ALT,AST,the hepatic level of ALT,AST,TC,TG,HDL-c,LDL-c,GSH-Px,SOD,MDA,CAT,and TNF-α,IL-1β,IL-6,CRP were determined by the kit and ELISA.The histopathological changes of the rat liver were examined by H&E,oil red O,and PAS staining.In vitro experiments,HepG2 cells were induced by FFA,and the level of TC,TG,HDL-c,LDL-c,GSH-Px,SOD,MDA,CAT,and TNF-α,IL-1β,IL-6,CRP in the cells were detected by kit and ELISA methods.Additionally,western blot,immunohistochemistry,and immunofluorescence were used to investigate the effects of EGK on the relevant proteins expression of SREBP-1c pathway(FAS,ACC,and SCD-1)in vivo and in vitro.Finally,by designing orthogonal experiments,the extraction process of EGK was optimized with the factors of solid-liquid ratio,ethanol concentration,extraction time,and extraction frequency as influencing factors,and the comprehensive scores of total flavonoid yield and extraction yield as evaluation indicators,the preparation process of EGK granules was optimized based on the factors of drug to auxiliary ratio,auxiliary material type,auxiliary material ratio,ethanol concentration,and the comprehensive scores of molding rate,angle of rest,moisture absorption rate,and bulk density as evaluation indicators.The experimental results showed that 8-weeks treatment with EGK has significantly decreased the level of blood glucose in rats,liver function indicators such as ALT,AST have been significantly improved,lipid accumulation indicators as TC,TG,LDL-C,HDL-C,oxidative stress indicators as SOD,GSH-Px,CAT,MDA,and inflammatory indicators as TNF-α,IL-6,IL-1β,CRP were significantly improved.Be consistent with above results,HE stainning showed that EGK improved the aggregation and infiltration of inflammatory cells in liver,and PAS staining and oil red O staining showed that EGK effectively improved glycogen accumulation and lipid accumulation in the liver.In addition,EGK significantly reduced lipid accumulation,and inhibited the oxidative stress,and inflammatory reactions in HepG2 cells induced by FFA.Immunohistochemical,immunofluorescence,and western blot results all showed that EGK could improve the abnormal expression of SREBP-1c,ACC,FAS,and SCD-1 proteins in vivo and in vitro.The optimization results of the preparation process of EGK granules show that the optimal extraction process for EGK is as follows: solid-liquid ratio is 1:20,ethanol concentration is 60%,extraction time is 2 h,and extraction times is thrice.The optimal preparation process for EGK granules is as follows: the ratio of drug to adjuvant is 1:1,the ratio of dextrin to sugar powder is 2:1,and ethanol concentration is 90%.After verification under the optimal extraction process conditions,the total flavonoid extraction rate is 85.28%,and the extract yield is 35.43%.Under the optimal preparation process conditions,the molding rate is 88.53%,the angle of repose is23.62 degrees,the moisture absorption rate is 6.30%,and the bulk density is 0.60 g/m L,indicating that the optimal process is stable and feasible.To sum up,EGK can dose-dependently improve liver dysfunction by modulating lipid accumulation,improving the imbalance of oxidative stress and inflammatory reaction in T2 DM combined with NAFLD rats,which suggesting that EGK has hypolipidemic,antioxidant,and anti-inflammatory effects and EGK has a good protective effect on the T2 DM combined with NAFLD rat.The in-vitro experiments results also indicate that EGK exhibit a good protective effect in FFA-induced HepG2 through improving the lipid disorder,oxidative stress and inflammation.As an important pathway to regulate lipid metabolism-SREBP-1c,EGK significantly improved the related protein expression of SREBP pathway in vitro and in vivo.Thus,we infer that the protective effect of EGK on T2 DM combined with NAFLD rats may be achieved by mediating the protein expression of SREBP-1c signaling pathway.Combined with our research on the preparation process of EGK granules,it provides a certain research foundation for the clinical application of EGK.