| Purpose:Based on TGF-β/ Smad signal pathway was used to explore the mechanism of the combination of Huangqi Gegen Decoction and Yuanzhi Decoction in inhibiting vascular EndMT in atherosclerotic rats with diabetes.Objective: To investigate the mechanism of inhibiting endothelial cell mesenchymalization(EndMT)in diabetic atherosclerotic rats based on the TGF-β/Smad signaling pathway by combining Huangqi Ge Gen Tang and Yuanzhi Tang.Material and method: 70 SPF-grade SD rats were randomly divided into blank control group,model group,positive control group,group 1 of the combined formula of Huangqi Ge Gen Tang and Yuan Zhi Tang,and group 2 of the combined formula of Huangqi Ge Gen Tang and Yuan Zhi Tang according to blood glucose.On the 29 th day of the experiment,the blank control group was given an intraperitoneal injection of citrate buffer;the remaining groups were given an intraperitoneal injection of 2% concentration of STZ to induce diabetic atherosclerosis in the rat model.After 49 days of continuous gavage,rat aortic tissues were subjected to HE staining,and serum glucose,CHO,TG,Hb A1 c,HDL-C,LDL-C levels were detected in each group;TGF-β,Vimentin,Twist1,Snail1 levels were detected by ELISA;liver SREBP-1c and The relative expression of rat aortic Vimentin,GLUT-4,AMPK and liver SREBP-1c m RNA was measured by realtime RT-PCR.The experimental data were analyzed using the statistical software SPSS 17.0.Results:1.Effects on blood glucose and blood lipids in rats(1)(1)Effects on blood glucose in rats: before modeling and after 28 days of high-fat diet,the groups no difference in blood glucose(P > 0.05).On the 49 th day of treatment,the model group had increased blood glucose compared to the blank control group(P < 0.01).Compared with the model group,the blood glucose in the positive control group,the combined formula1 group and the combined formula 2 group decreased significantly(P < 0.01).(2)Effects on Hb A1 c in rats: compared with the blank control group,Hb A1 c was higher in the model group than in the blank control group(P < 0.01).Hb A1 c was lower in the model group than in each of the administered groups(P < 0.01).Compared with the combined formula 1 group,the combined formula 2 group was more effective in reducing serum Hb A1 c in rats(P < 0.05).(2)Effects on blood lipids in diabetic atherosclerotic rats:(1)Effects on TG in rats: The TG of the model group was higher than that of the blank control group(P < 0.05).Compared with the model group,the serum TG in the positive control group,the combined formula 1group and the combined formula 2 group was significantly lower(P < 0.05).(2)Effects on serum CHO in rats: CHO was high in the model group versus the blank control group(P <0.05).Compared with the model group,all the administered groups could significantly reduce CHO(P < 0.01).(3)Effect on HDL-C in rats: HDL-C was significantly lower in the model group compared with the blank control group(P < 0.05).Compared with the model group,the serum HDL-C of rats in the positive control group,the combined formula 1 group and the combined formula 2 group increased significantly(P < 0.05).(4)Effects on LDL-C in rats:LDL-C was significantly lower in the model group compared with the blank control group(P< 0.05).LDL-C was significantly lower in each administration group compared to the model group(P < 0.05).2.Effect on the histomorphology of aorta of diabetic atherosclerotic rats:The aortic cells in the blank control group were neat,morphologically clear and structurally intact,with small lipid nuclei,and no plaque formation was found;the aortic cells in the model group were disordered,morphologically blurred and structurally incomplete,with large lipid nuclei,a large number of foam cells,lipid crystals and unstable plaques;the aortic cells in the positive control group and the combined formula 1 group were mildly more disordered,morphologically blurred and structurally intact than those in the model group.The aortic cells in the positive control group and the combined formula 1 group were slightly disordered,the morphology was slightly blurred,the structure was slightly intact,the lipid nuclei were large,foam cells were visible,and the plaque was slightly unstable;the aortic cells in the combined formula 2 group were neater,the morphology was clearer,the structure was more intact,the lipid nuclei were small,and the plaque was stable.3.Effects on the expression of Vimentin,Glut4,AMPK and hepatic SREBP-1C m RNA in rat aorta(1)Effects on the expression of Vimentin in rat aorta: Vimentin was significantly increased in the model group compared with the blank control group(P < 0.01).Vimentin was significantly decreased in g each administration group compared with the model group(P <0.05).The Vimentin expression level in the positive control group was significantly higher than that in the combined formula 1 group(P < 0.05).Compared with the combined formula 1group,the combined formula 2 group had a better effect in reducing Vimentin in rat aorta,and the difference was statistically significant(P < 0.05).(2)Effect on the expression of Glut4 in rat aorta: Glut4 was significantly reduced in the model group compared to the blank control group(P < 0.01).Compared with the model group,aortic Glut4 was significantly increased in the positive control group,the combined formula 1group and the combined formula 2 group(P < 0.05).Glut4 was significantly lower in the positive control group compared with the Hopping Formula 2 group(P < 0.05).Compared with the Hopping Formula 1 group,the Hopping Formula 2 group had a better effect in elevating rat aortic Glut4(P < 0.05).(3)Effects on AMPK expression in rat aorta: compared with the blank control group,AMPK expression was decreased in the model group(P < 0.01).The expression of AMPK was significantly increased in the g-administered group compared with the model group(P <0.05).Compared with the Hopping Formula 1 group,the Hopping Formula 2 group had a better effect in enhancing AMPK in rat aorta,and the difference was statistically significant(P<0.05).(4)Effect on the expression of SREBP-1C in rats: compared with the blank control group,SREBP-1C was significantly increased in the model group(P < 0.01).Compared with the model group,positive control group,combined formula 1 group,combined formula 2group rat liver SREBP-1C m RNA significantly decreased(P < 0.05).Compared with the positive control group,the SREBP-1C level was increased in the Hopping Formula 1 and Hopping Formula 2 groups(P < 0.05).Compared with the Hopping Formula 1 group,the Hopping Formula 2 group was more effective in reducing SREBP-1C m RNA in rat liver,and the difference was statistically significant(P < 0.05).4.Effects on the expression of serum Vimentin,TGF-β,Twist1 and Snail1 m RNA in rats(1)Effects on serum Vimentin m RNA expression in rats: Vimentin was increased in the model group compared to the blank control group(P < 0.01).Serum Vimentin was significantly lower in the positive control,combined formulation 1 and combined formulation2 groups compared with the model group(P < 0.01).Compared with the positive control group,the Vimentin level in the Hopping Formula 1 group was significantly increased(P <0.01).Compared with group 1,group 2 was more effective in reducing serum Vimentin in rats(P < 0.05).(2)Effects on serum TGF-β m RNA expression in rats: TGF-β levels were elevated in the model group compared to the blank control group(P < 0.01).Compared with the model group,TGF-β was significantly lower in the positive control group,combined formulation 1 group,and combined formulation 2 group(P < 0.01).Compared with the positive control group,the TGF-β levels in the Hopping Formula 1 and Hopping Formula 2 groups were significantly increased(P < 0.01).The serum TGF-β m RNA expression was significantly lower in the Hopping Formula 2 group compared with the Hopping Formula 1 group(P < 0.05).(3)Effects on serum Twist1 m RNA expression in rats: Twist1 was elevated in the model group compared with the blank control group(P < 0.01).Twist1 was decreased in the positive control group,combined formula 1 group and combined formula 2 group compared to the model group(P < 0.01).Twist1 levels were elevated in the combined formula 1 group and combined formula 2 group compared with the glycoprotein granule group(P < 0.01).The expression of Twist1 m RNA in serum of rats in the Hopping Formula 2 group was significantly lower compared with that in the Hopping Formula 1 group(P < 0.05).(4)Effects on serum Snail1 m RNA expression in rats: Compared with the blank control group,Snail1 levels were elevated in the model group(P < 0.01).Snail1 levels were significantly lower in the positive control,combined formulation 1 and combined formulation2 groups compared with the model group(P < 0.01).Compared with the positive control group,the Snail1 level in the Hopping Formula 1 group were increased(P < 0.01).and decreased in the Hopping Formula 2 group(P < 0.05).The serum Snail1 m RNA expression was significantly lower in the Hopping Formula 2 group compared with the Hopping Formula1 group(P < 0.05).5.Effects on the expression of Smad2,Smad3 and hepatic SREBP-1C in rat aorta(1)Smad2 levels were increased in the model group compared to the blank control group(P < 0.05).Compared with the model group,the aortic Smad2 levels were significantly lower in the combined formula 1 group,and the combined formula 2 group(P < 0.05).(2)Compared with the blank control group,aortic Smad3 was significantly elevated in the model group(P < 0.05).Compared with the model group,the aortic Smad3 level was significantly lower in the Hopping Formula 1 and Hopping Formula 2 groups(P < 0.05).(3)SREBP-1C levels were increased in the model group compared to the blank control group(P < 0.01).Compared with the model group,the aortic SREBP-1C levels were significantly lower in the Hopping Formula 1 and Hopping Formula 2 groups(P < 0.05).Compared with the positive control group,the SREBP-1C level was significantly increased in the Hopping Formula 1 group(P < 0.05).Conclusion:1.The combination of Huangqi Gegen Decoction and Yuanzhi Decoction can effectively inhibit the interstitial formation of aortic endothelial cells in atherosclerotic rats with diabetes.In the aspect of inhibiting endothelial cell interstitial fibrosis,the effect of Hefang 2 group was better.2.The mechanism of the combination of Huangqi Gegen Decoction and Yuanzhi Decoction in inhibiting ENDMT in atherosclerotic rats with diabetes may be related to the activation of TGF-β/ Samd signal pathway.That is,by lowering TGF-β And then inhibit the expression of downstream Smad2 and Smad3,control the formation of Smad trimer,inhibit the expression of ENDMT related genes such as Snail1 and Twist1,and thus inhibit the process of ENDMT.3.The combination of Huangqi Gegen Decoction and Yuanzhi Decoction can effectively reduce the blood glucose and lipid levels of atherosclerosis rats with diabetes.In terms of regulating blood sugar and blood lipids,the effect of the two groups is better.Its regulatory mechanism may be related to the activation of AMPK signaling pathway.That is,regulate glucose and lipid metabolism by up-regulating the expression of AMPK m RNA,thereby increasing the downstream GLUT4 and inhibiting the expression of SREBP-1C.The effect is better in adjusting Hb A1 C combination formula group 2.4.The combination of Huangqi Gegen Decoction and Yuanzhi Decoction can prevent and treat atherosclerosis in diabetes by improving glucose and lipid metabolism and inhibiting the process of aortic ENDMT. |
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