Study On The Changes Of Glucose Transport And Energy Metabolism In Spleen-yin Deficiency Rats And The Effect Of ZiBu PiYin Recipe

Objective:To investigate the changes of gastrointestinal glucose transport and mitochondrial energy metabolism in rats with spleen-yin deficiency,and to observe the interventional effects of nourishing spleen-yin prescription.Methods:Healthy SPF-grade Wistar male rats were first fed adaptively for 3 days,and then randomly divided into two groups:6 in the normal group and 14 in the model group.Classical compound factors were used to build the model,that is,a rat model of spleen-yin deficiency was established by using a diet that was not moderate,overworked and combined with yin medicine.The modeling time is 38 days,divided into 3 time periods:?The 1-14th day is the model of spleen deficiency,full food 1 day+fasting 2 days,that is,three days as a cycle,and swim every day exhaust.?On the 15th to 24th days,the spleen and yin deficiency model was established.The rats with the spleen and qi deficiency model in the model group were classified into the spleen and yin deficiency model.?The 25th to 38th day was the intervention stage of nourishing spleen yin prescription:the model group was randomly divided into spleen yin deficiency group and nourishing spleen yin prescription group,6 rats in each group.The Zibu Piyin Recipe group was given intragastrically 1 mL/100 g of the Zibu Piyin Recipe,and the normal and spleen-yin deficiency groups were given the same volume of saline.Gastrointestinal tissues were obtained by Western blotting and qPCR technology to detect the expression of glucose transporter protein and verify the intervention effect of nourishing spleen and yin prescriptions;ELISA method was used to detect skeletal muscle mitochondrial respiratory chain complex and ATP synthase activity to reflect changes in mitochondrial energy metabolism.Result:1.Changes of Glucose Transporters(GLUTs)in Spleen-Yin Deficiency Rats and Intervening Effects of Nourishing Spleen-Yin Recipe(1)Western blotting method was used to detect the expression of GLUT1 and GLUT4 protein in rat stomach:Compared with the normal group,the expression levels of GLUT1 and GLUT4 protein in the stomach of the spleen-yin deficiency group were significantly reduced(P<0.001,P<0.01);In the spleen-yin deficiency group,the expressions of gastric GLUT1 and GLUT4 protein were up-regulated in the nourishing spleen-yin prescription group(P<0.001,P<0.05).(2)Western blotting was used to detect the expression of GLUT1 and GLUT5 in the jejunum of rats:Compared with the normal group,the expression levels of GLUT1 and GLUT5 in the jejunum of the spleen-yin deficiency group were reduced(P<0.05);compared with the spleen-yin deficiency In the group,the jejunum GLUT1 and GLUT5 protein expressions of the nourishing spleen yin prescription group were all up-regulated(P<0.05).(3)qPCR method to detect the changes of GLUT1 and GLUT4 mRNA expression in rat stomach:Compared with the normal group,rats in the spleen-yin deficiency group had lower gastric GLUT1 mRNA expression(P<0.01),and GLUT4 mRNA expression had a downward trend;compared with the spleen-yin In the deficiency group,the expressions of GLUT1 and GLUT4 mRNA in the nourishing spleen yin prescription group were increased(P<0.05);compared with the normal group,the expressions of GLUT1 and GLUT4 mRNA in the nourishing spleen yin prescription group were also increased(P<0.05).(4)qPCR method to detect the expression changes of GLUT1 and GLUT5 mRNA in the jejunum of rats:Compared with the normal group,the expression of GLUT1 and GLUT5 mRNA in the jejunum of the spleen-yin deficiency group was reduced(P<0.05);compared with the spleen-yin deficiency group The expression of GLUT1 and GLUT5 mRNA in the nourishing spleen yin prescription group increased significantly(P<0.01);compared with the normal group,the expression of GLUT 1 mRNA in the nourishing spleen yin prescription group increased(P<0.05),and the expression of GLUT5 mRNA increased.High trend.2.Changes of Mitochondrial Energy Metabolism in Spleen-Yin Deficiency Rats and Intervention Effect of Nourishing Spleen-Yin Recipe(1)Activity detection of skeletal muscle mitochondrial respiratory chain complexes ?,?,?,and ?:Compared with the normal group,the skeletal muscle mitochondrial respiratory chain complexes ?,?,?,and ? activities increased P<0.01);After the intervention of nourishing spleen yin prescription,the activity of mitochondrial respiratory chain complexes ?,?,? and ? in spleen yin deficiency group was significantly reduced compared with that in spleen yin deficiency group(P<0.01),no significant difference compared with the normal group.(2)Detection of mitochondrial ATP synthase activity in skeletal muscle:Compared with the normal group,the mitochondrial ATP synthase activity in the skeletal muscle of the spleen-yin deficiency group was significantly increased(P<0.01);compared with the spleen-yin deficiency group,it nourished the spleen The mitochondrial ATP synthase activity of the yin prescription group decreased(P<0.01),and there was no significant difference compared with the normal group.Conclusion:1.The decrease of GLUT1,GLUT4 and jejunal GLUT1,GLUT5 protein and mRNA expression in rats with spleen-yin deficiency may be one of the important factors leading to digestive and absorption disorders of gastrointestinal glucose and other nutrients.Nourishing spleen and yin prescription can improve the glucose transport ability of spleen-yin deficiency rats by up-regulating the expression of GLUT1,GLUT4 and jejunum GLUT1,GLUT5 protein and mRNA in rats.2.In the state of spleen-yin deficiency,rats' mitochondrial energy metabolism increases abnormally.Nourishing spleen-yin prescription can reduce the abnormal activity of mitochondrial respiratory chain complex,weaken ATP synthesis,and improve mitochondrial energy metabolism to reduce spleen-yin deficiency syndrome on skeletal muscle mitochondria.The damage thus achieves a protective effect on mitochondrial function and structure.