| ObjectiveUsing ZDF rats who was induced by high fat diet as T2DM rat model,To investigate the effects of Gegen QinLian Decoction?GQD?on intestinal microflora and LPS/TLR-4/NF-?B signaling pathway in the ZDF rats for metabolic endotoxemia of T2DM,and to analyze its mechanism of action on T2DM.Methods8 male ZDF?fa/+?rats and 24 male ZDF?fa/fa?in rats fed normal diet 3 days after adaptation,ZDF?fa/+?rats were given normal diet,ZDF?fa/fa?fed with high fat diet for 21 days,21 days after the blood glucose was measured,random blood samples were more than16.7mmol/L that model was induced successfully.The model was successfully induced and 8male ZDF?fa/+?rats as normal control group(0.9%NaCl,10ml·kg-1),24 male ZDF?fa/fa?rats were randomly divided into model group(0.9%NaCl,10ml·kg-1),metformin group(0.16g·kg-1),GQD group(13.8g·kg-1),by gavage.After the drug was given,the normal control group was given to the normal diet,and the model group and the administration group were still fed with high fat feed.1 times a day,record the daily amount of drinking water,food intake,observe the spirit,hair and so on.Weight and fasting blood glucose were measured weekly,and the medicine was administered for 8 weeks.After 8 weeks of administration,the feces of rats were collected by the tail reflection method in the aseptic cryopreservation tube,and stored at-80?.After 14 hours of fasting,the OGTT test was performed.Automatic biochemical measurement of TG,TC,HDL-C,LDL-C,FFA,FINS.ELISA was used to detect the amount and activity of LCAT and LPL in serum,and the levels of protein?Zonulin?,two amine oxidase?DAO?,lipopolysaccharide?LPS?and fecal LPS.Calculate AUC,IRI,ISI.The pathological changes of the pancreas,liver and ileum were observed by HE staining.The changes of intestinal flora were analyzed by 16S rDNA sequencing.The protein expression of TLR4 and NF-?B in the ileum,pancreas and liver tissues was detected by WB.The expression of TLR4 and NF-?B protein in the ileum,pancreas and liver were observed by immunohistochemistry.Results1.Sugar and lipid metabolism results:The weight and FBG of the model group increased significantly at all stages to compare with the normal group?P<0.01?.the body weight and FBG in the GQD group decreased significantly at all stages to compare with the model group,?P<0.05?.Levels of model group in TG,TC,HDL-C,LDL-C,FFA,FINS,HbA1C were significantly increased to compare with the normal group?P<0.01?;Levels of GQD group inTG TC,LDL-C has decreased,but the difference was not statistically significant to compare with the model group?P>0.05?,level of in HDL-C,FFA,FINS,Hb A1C were ont significantly decreased to compare with the model group?P<0.05?;levels of TC and TG in GQD group were decreased to compare with metformin group?P<0.05?,but no statistically significant differences in the level of LDL-C to compare with metformin group?P>0.05?.The IRI in the model group was significantly higher to compare with the normal group?P<0.01?.the IRI of metformin group and GQD group decreased significantly to compare with the model group?P<0.01?.the ISI in the model group decreased significantly to compare with the normal group?P<0.01?.the difference of ISI in metformin group was not statistically significant to compare with the model group?P>0.05?,and the ISI of GQD group was significantly higher to compare with the model group?P<0.01?.the level and the activity of LCAT in the model group was increased to compare with the normal group?P<0.05?.the level and the activity of LCAT in GQD group were decreased to compare with the model group?P<0.05?.the level and activity of LPL in the model group were decreased to compare with the normal group?P<0.01?.the level and activity of LPL in the GQD group were increased to compare with the model group?P<0.05?.HE results showed that,the liver and pancreatic tissue of the rats in the model group were severely damaged to compare with the normal group.the liver and pancreatic tissue of GQD group were less damaged to compare with the model group.2.Intestinal flora results:the proportion of Gram-negative bacteria of the model group was significantly higher to compare with the normal group?P<0.05?.the proportion of Gram-negative bacteria in GQD group decreased significantly to compared with the model group?P<0.05?.the level of Prevotella and Christensenella in the model group decreased significantly to compared with the normal group?P<0.05?.the level of Prevotella and Christensenella in GQD group increased significantly to compared with the model group?P<0.05?.3.metabolic endotoxemia results:the serum levels of Zonulin and DAO,model group were significantly increased to compare with the normal group?P<0.01?;GQD group were significantly decreased to compare with the model group?P<0.05?;the serum Zonulin,GQD group decreased significantly to compare with the metformin group?P<0.01?.the LPS in the stool and serum,the model group was increased significantly to compare with the normal group?P<0.01?;GQD group was decreased significantly to compare with the model group?P<0.01?;GQD group was decreased significantly to compare with the metformin group?P<0.05?.The results of WB showed that the relative expression of TLR-4 and NF-?B in the ileum,liver and pancreas,the model group was significantly higher than that in the normal group?P<0.01?;metformin group and GQD group decreased to varying degrees to compare with the model group?P<0.05?.Immunohistochemical results showed that was compared with the normal group,the average optical density of TLR-4 and NF-?B in the ileum,liver and pancreas tissues of model group increased significantly?P<0.01?.the average optical density of TLR-4 and NF-?B in ileum,liver and pancreas tissues of metformin group and GQD group was decreased to varying degrees to compared with the model group?P<0.05?.Conclusion1.Gegen Qin Lian decoction can effectively control the body weight of ZDF rats,improve the metabolism of sugar and improve the metabolism of some fat.2.Gegen Qin Lian decoction can reduce the proportion of Gram-negative bacteria in the intestinal flora of ZDF rats,and increase the abundance of Prevotella and Christensenella.3.Gegen Qin Lian decoction can effectively improve the"metabolic endotoxemia"in the body of ZDF rats. |
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