| Objective: A systematic study on the levels of urinary regulatory hormones,hepatic glucose ATP and Na-K-ATPase activities,the expression of various water transporters in the liver,the content of water transporter mRNA,and the level of phosphorylation in rats with dampness block and middle jiao syndrome.To explore the effect of syndrome of dampness in middle Jiao on the activity of Na-K-ATPase in liver,the effect of urine production regulating hormone,water transport protein and the intervention of Pingwei Powder,and to explore whether Pingwei Powder can regulate the imbalance of urine production regulation hormone by affecting Na-K-ATPase activity.Further affect the liver water-sugar co-transporter to regulate the state of water and liquid metabolism,and reveal some of the molecular mechanism of cell water and liquid metabolism imbalance caused by syndrome of dampness in middle Jiao and the role of Pingwei Powder in regulating cell transmembrane water transport.Method: Spleen and stomach dampness obstruction after making model,through the activity of Na K ATPase in liver was determined by chemical colorimetry after modeling in middle Jiao,glucose content in liver was detected by peroxidase method,and ATP content in liver was detected by high performance liquid chromatography(HPLC).Determination of SGLT1,GLUT2,NKCC1 in liver tissue by enzyme-linked immunosorbent assay(ELISA).The content of mRNA in liver tissue was determined by Polymerase Chain Reaction(PCR).The phosphorylation level of SGLT1,GLUT2,NKCC1 in liver tissue was determined by western blot(WB)method.Results:1 Evaluation of the Syndrome of dampness in middle Jiao model:(1)Observation through macroscopic signs,rats with dampness obstruction and middle Jiao syndrome rats significantly reduce the weight,food intake and drinking water,and poor mental state.(2)The objective index detection,gastric emptying disorder,content of serum D-xylose absorption,hepatic glucose accumulation and ATP production decreased Na-K-ATPase activity.(3)The immune expression ofvarious water transporters in the liver tissue of the spleen and stomach was abnormal.(4)After taking Chinese medicine,feeding,drinking water,weight,gastric residual rate,the intestinal propulsion rate,Speculated that Immune function,serum D-xylose absorption,liver glucose utilization,Reduction of ATP formation and reduction of sodium pump activity.It was speculated that the syndrome of dampness in middle Jiao model rats building success.2 rine production and regulation of hormone content:After animal model building,rats with dampness obstruction and middle Jiao syndrome rats compared with no module,the contents of renin,angiotensin,antidiuretic hormone,endothelin and epinephrine decreased,aldosterone and potassium natriuretic peptide increased,and atrial natriuretic peptide did not change in Rats with dampness obstruction and middle Jiao syndrome.The levels of renin,angiotensin and atrial natriuretic peptide in the rat self-repair group were higher than those in the Rats with dampness obstruction and middle Jiao syndrome,aldosterone,potassium diuretic peptide,antidiuretic hormone,endothelin and adrenaline were lower than those in Rats with dampness obstruction and middle Jiao syndrome.The contents of renin,angiotensin,aldosterone and atrial natriuretic peptide in the high,middle and low dose groups were higher than those in the rat self-repair group,and the levels of antidiuretic hormone and adrenaline in the medium and low dose groups were lower than those in the Rat self-repair group.The content of endothelin in medium dose group of Pingwei Powder was higher than that of Rat self-repair group,the content of potassium diuretic peptide in medium dose group of Pingwei Powder was higher than that of self repairing group,the content of endothelin in low dose group of Pingwei Powder was lower than that of self repairing group,and that of Pingwei Powder was high.The content of potassium diuretic peptide in low dose group was lower than that in self repairing group.3 Expression of liver water transport protein,content of mRNA and phosphorylation activity:(1)Changes of mRNA content and phosphorylation activity of water-sugar co-transporterCompared with the normal group,the expression of SGLT1 protein was up regulated in the liver of rats with damp-trapped spleen and stomach,the content of SGLT1 decreased,and the phosphorylation activity of SGLT1 decreased.After the recovery,the expression of SGLT1 protein in the liver of the recovery group was up-regulated and the content of SGLT1 decreased,and the phosphorylation activity of SGLT1 decreased.Compared with the natural recovery group,the expression of SGLT1 in the liver of the traditional Chinese medicine group was up-regulated,the content of SGLT1 increased,the content of mRNA was increased,the phosphorylation activity of SGLT1 increased and decreased.After dampness of spleen and stomach,compared with blank group,the expression of GLUT2 protein in the liver of rats with dampness trapped spleen and stomach showed that the content of GLUT2 decreased and the activity of phosphorylation decreased,while that of rats with dampness trapped spleen and stomach was compared with that of rats with dampness trapped spleen and stomach.After natural recovery,the expression of GLUT2 protein in the liver decreased,the content of mRNA was increased,the phosphorylation activity increased,and compared with the recovery group,the protein expression of GLUT2 in liver is down-regulated after different doses of Pingwei Powder.The content of GLUT2 is high and the content of GLUT2 is increasing.The activity of GLUT2 phosphorylation in liver of low dose group of Pingwei San was increased,and the activity of phosphorylation of GLUT2 in liver of low dose group was higher than that of medium dose group.(2)The content of immune expression,the content of mRNA and the change of phosphorylation activity of water salt cotransporterAfter modeling,compared with the control group,the expression of NKCC1 protein in the liver of rats with dampness trapped spleen and stomach was up-regulated,the content of NKCC1 was increased,the content of nkCC1 was increased and the phosphorylation activity of NKCC1 was decreased.Compared with the rats with dampness trapped spleen and stomach syndrome,the down-regulation of NKCC1 protein expression in the liver after recovery could decrease the content ofNKCC1 and increase the phosphorylation activity of NKCC1,as compared with the natural recovery group.The protein expression of NKCC1 was up-regulated in high dose group of Pingwei Powder,and down-regulated in low dose group and medium dose group of Pingwei Powder.The content of NKCC1 was increased in different dosage groups,and the content of NKCC1 was higher in Pingwei San group.The activity of NKCC1 phosphorylation in liver of middle dose group was decreased,while that of NKCC1 in low dose group was higher than that in low dose group.(3)The content of aquaporin immuno-expression and the change of mRNA content and phosphorylation activity of aquaporin.The results showed that after dampness trapped spleen and stomach,the expression of aquaporin1 and aquaporin4 in liver was increased,and the content of aquaporin1 and aquaporin4 in liver were decreased,and the activity of phosphorylation was decreased after taking different doses of Pingwei Powder.The immunoexpression of aquaporin1 and aquaporin4 decreased,and the content and phosphorylation activity of aquaporin1 and aquaporin4 increased.The results showed that after dampness obstruction and middle Jiao syndrome,the expression of aquaporin1 and aquaporin4 in liver was increased,and the content of aquaporin1 and aquaporin4 in liver were decreased,and the activity of phosphorylation was decreased after taking different doses of Pingwei Powder.The immunoexpression of aquaporin1 and aquaporin4 decreased,and the content and phosphorylation activity of aquaporin1 and aquaporin 4 increased.Conclusion:Syndrome of dampness in middle Jiao model not only affect the metabolism hormones of renin,angiotensin,aldosterone,endothelin,antidiuretic hormone,atrial natriuretic peptide and potassium uridine peptide,on the other hand,it leads to abnormal expression of liver water transport protein..The expression of liver water-sugar co-transporter SGLT1 and water-salt co-transporter NKCC1 was up-regulated,the expression of liver water-sugar co-transporter GLUT2 was down-regulated,and the contents of liver water-sugar co-transporter SGLT1 and GLUT2 were decreased.The content of mRNA in liver SGLT1 and GLUT2、NKCC1was decreased and reduced phosphorylation levels of GLUT2、NKCC1 and SGLT1 in the liver.Increased hepatic glucose content decreased ATP generation,Na-K-ATP enzyme activity decreased.Pingweisan can improve energy metabolism,enhanced Na-K-ATP enzyme activity in the liver,regulating serum fluid metabolism in regulating hormone balance,regulate the amount of urine cell and electrolyte balance,while the expression by regulating the liver water transport protein content and phosphorylation activity,ion balance inside and outside cells,so that the body of the whole water metabolism balance. |
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