| Objective:To research the effects of different extracts of Zhuang-Medicine Cardiospermum halicacabum L.on hypoglycemic therapeutical effect,oxidative stress and inflammatory injury of Mice with Type 2 Diabetes Mellitus induced by high-fat and high-sugar diet(HFD)combined with streptozotocin(STZ),and to preliminarily explore its possible action mechanism,so as to provide experimental basis for its clinical development and application.Method:1.Model establishment,grouping and administration methods: In this study,70 healthy Kunming male mice(KM)were used to establish diabetes mellitus model.Firstly,they were fed adaptively for 3 days.Ten mice were randomly selected into the normal group and fed with standard pellet feed.The remaining 60 mice were fed with high-fat and high-sugar feed for 40 days.After40 days,STZ(60mg·kg-1)was injected into the abdominal cavity of high-sugarand high-fat mice for 2 times.Each time interval was 3 days.One week later,fasting blood glucose(FBG)was detected after fasting and water was allowed for 12 hours.Mice with FBG greater than 11.1 mmol·L-1 were selected as successful mouse models of type 2 diabetes.50 model mice were randomly divided into 5 groups according to FBG value: model group,metformin group,Ethanol extracted substance of Cardiospermum halicacabum L.(EESCHL)group,Ethyl acetata extracted substance of Cardiospermum halicacabum L.(EAESCHL)group and N-butanol extracted substance of Cardiospermum halicacabum L.(NBESCHL)group.The mice continued to be fed with normal diet in the normal group,while the mice continued to be fed with high-fat and high-sugar diet in the model group.Metformin group and different extracts of Cardiospermum halicacabum L.groups were given corresponding concentrations of Metformin and different extracts of Cardiospermum halicacabum L.,once a day.2.During the period,the mice were observed daily and their body weight changes were recorded.FBG was detected once a week by tail vein blood collection and recorded.On the 14 day,oral glucose tolerance test(OGTT)was carried out after fasting and water was allowed for 12 hours,After the collection of serum samples,organ samples were taken and organ index was calculated.Samples of pancreas and liver tissues of mice were collected in each group and sectioned after fixation to observe and compare the histopathological morphology changes.3.The level of Serum total cholesterol(TC),triglyceride(TG),high density lipoprotein-cholesterol(HDL-C),low density lipoprotein-cholesterol(LDL-C),albumin(ALB),globulin(GLB),total protein(TP),blood urea nitrogen(BUN),creatinine(CR),alanine aminotransferase(ALT),aspartateaminotransferase(AST),alkaline phosphatase(ALP)were detected by automatic biochemical analyzer.4.The changes of fasting insulin(FINS),glycosylated hemoglobin(HbAlc),inflammatory factor interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA),then calculate the insulin resistance index(IRI).5.The levels of oxidative factor superoxide dismutase(SOD),Maleic dialdehyde(MDA),glutathione(GSH)and Glutathione-Px(GSH-Px)in liver tissue were detected by enzyme-labeled kit.Results:1.The effect on general symptoms: Compared with the model group,with the prolongation of administration time,the mental state of mice were improved in different extracts of Cardiospermum halicacabum L.groups,and the amount of diet,drinking water and urine decreased,which improved the clinical symptoms of type 2 diabetes mellitus mice to a certain extent.2.The effect on blood glucose: Different extracts of Cardiospermum halicacabum L.have a decreased effect on blood glucose in type 2 diabetic mice.On the 7th day of administration,the FBG of the mice in NBESCHL group had decreased significantly(P<0.05).On the 14 th day of administration,the FBG of the mice in each group was significantly decreased(P<0.01).The OGTT test showed that the blood glucose of each extract group decreased at 30 min,60min and 120 min,and the blood glucose value decreased rapidly at 120 min,and the overall decline trend was slightly weaker than that of the metformin group.Compared with the model group,HbAlc and IRI in each group of mice were compared.HbAlc was significantly decreased in NBESCHL group and inEAESCHL group(P<0.05 or P<0.01),and the IRI decreased significantly in each drug-administered group(P<0.05).The long-term glycemic control of type2 diabetes was obvious.effect.3.The effect on organ index : Compared with the model group,the thymus and spleen index of mice had little change in different extracts of Cardiospermum halicacabum L.groups,while the weight of liver and kidney and the index of mice decreased significantly in EAESCHL group and NBESCHL group(P < 0.01).4.Effects on blood lipids: Compared with the normal group,the TG,TC,LDLC and HDLC of the model group showed obvious abnormalities(P<0.01),and the TG,TC,LDLC and HDLC of the different extracts of Cardiospermum halicacabum L.groups showed the downward trend,but the difference was not significant(P>0.05).5.Effects on liver and kidney function: Compared with the normal group,the AST,ALT,and ALP of the model group were significantly increased(P<0.01),and the AST,ALT,and ALP of the different extracts of Cardiospermum halicacabum L.groups were different degrees.The decrease of AST,ALT and ALP in NBESCHL group was significantly decreased(P<0.05).The ALB of the model group was significantly decreased(P<0.01),and the GLB and TP decreased.The levels of ALB,GLB and TP in type 2 diabetic mice were increased.The ALB levels in EAESCHL group and NBESCHL group were significantly increased(P<0.05).The renal function indicators showed that the BUN and CR of the mice in each experimental group did not change much.6.Effects on tissue structure: HE staining results showed that compared with the model group,the damage of the pancreas and liver tissue structure of the mice in each of the extracts of Cardiospermum halicacabum L.was significantly alleviated.7.The effect on liver oxidative factors: Compared with the normal group,the SOD of the model group was significantly decreased(P<0.05),MDA was significantly increased(P<0.01),and the SOD of the different extracts of Cardiospermum halicacabum L.groups was increased,while the MDA was decreased.There was a decrease in SOD in EAESCHL group and NBESCHL group(P<0.05),and a significant decrease in MDA in EESCHL group and NBESCHL group(P<0.05).The model group had a slight decrease in GSH-PX,The decrease was significant in GSH.(P<0.05).The GSH-PX and GSH of the different extracts of Cardiospermum halicacabum L were increased to different extents.The GSH of EAESCHL group was increased(P<0.01),and the GSH of NBESCHLgroup and the metformin group were also increased Significantly(P <0.05).8.Effects on inflammatory cytokines: the different extracts of Cardiospermum halicacabum L.decreased the levels of IL-6 and TNF-α in serum,but there was no significant difference(P > 0.05).Conclusion:Different extracts of Cardiospermum halicacabum L.can improve the general clinical symptoms of type 2 diabetes mellitus mice,such as the improvement of glucose metabolism in the body,the reduced of insulin resistance,the improvement of liver function,and protective effect on the damage of islet cells and liver cells.In addition,different extracts can also reduce oxidative damage in type 2 diabetes mellitus mice,improve their antioxidant capacity.It is speculated that the hypoglycemic mechanism of Cardiospermum halicacabum L.may be related to the regulation of glucose metabolism,the protection of islet function and the antioxidant activity of theorganism.Among the three different extracts from the Cardiospermum halicacabum L.,NBESCHL group has the more obvious therapeutic effect,which needs to be further explored and developed. |
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