| Objective: Study of kidney deficiency physique,kidney deficiency syndrome in rats growth and development status,and based on parotid water channel protein change,from the point of view of water metabolism,exploring the molecular mechanism of kidney associated with saliva for clinical illness associated with kidney stores like salivary fluid micro differentiation and non-invasive diagnostic technique foundation,provide objective basis for early treatment is associated with kidney disease.Methods: According to the theory of "fear of hurting the kidney",the rat model of congenital deficiency kidney deficiency constitution was established by selecting the newly sexually mature rats and using the modeling method of "cat scare the rat".On this basis,the rat model of kidney deficiency constitution was continuously injected with "hydrocodelasone injection",so as to establish the animal model of kidney deficiency syndrome.Combined with "the kidney stores essence,the main growth" "kidney in liquid for salivary" theory,observation records each general situation of rats,to detect the rat body weight,weight of parotid gland,saliva flow rate,osmotic pressure,saliva TNF alpha saliva,AQP1 protein in the parotid gland,AQP5 protein and nf-kappa Bp65 protein and Bp-p65 nf-kappa protein related indicators of change,explore the internal mechanism of kidney associated with salivary.(1)General observation: During the experiment,rats in the normal control group were energetic,fond of climbing,active,active,fast,fast growth and development,high body weight,no hair loss,thick fur,strong appetite,normal water consumption,normal urine and feces.The body weight of rats in the kidney deficiency group was slightly lighter than that in the normal control group,their activities were reduced,and their spirits were depressed.Some rats had hair loss,poor hair luster,less water and food intake than the normal control group,more urine,and soft stools,which basically matched the macroscopic manifestation of the kidney deficiency group.Rats in the kidney deficiency syndrome group were listless,timid and easily frightened,less active,like huddle and curl up,hair sparse and falling off,water and food intake significantly reduced,urine volume was much,stool was not formed,which was consistent with the macro manifestation of kidney deficiency syndrome.(2)Body weight at the end of modeling: Compared with the normal control group,the body weight of the kidney-deficiency constitution group and kidney-deficiency syndrome group decreased,both of which had statistical significance(P < 0.01).(3)Parotid gland weight: Compared with normal control group,the weight of parotid gland of rats in kidney deficiency constitution group and kidney deficiency syndrome group showed a decreasing trend,with statistical significance P < 0.05).(4)Saliva flow: Compared with the normal control group,the amount of saliva secretion in the kidney deficiency group was decreased,with statistical difference(P <0.05),while the amount of saliva secretion in the kidney deficiency group was significantly decreased,with statistical difference(P < 0.01).(5)Saliva osmotic pressure: Compared with the normal control group,the saliva osmotic pressure of rats in the kidney deficiency group was decreased,with statistical significance(P < 0.05),while the saliva osmotic pressure of rats in the kidney deficiency group was significantly decreased,with statistical significance(P < 0.01).(6)Salivary TNF-α ELISA: Compared with normal control group,saliva TNF-αincreased in renal deficiency constitution group and renal deficiency syndrome group,and there was statistical significance in renal deficiency syndrome group(P < 0.05).(7)All landowners parotid HE staining: normal control group rats neat parotid gland flocculus,a clear demarcation between the interlobular septa,acinar cell morphological rules,the normal structure and abundant cytoplasm,nuclear,no shrinkage,glandular cell at the top of the secretory granules,leap tube and catheter lumen,neat wall cells,capillary endothelial proliferation,squamous epithelial cells,not to the tube cavity is prominent;No obvious inflammation was observed.In the renal deficiency group,there were more acinar cells with reduced cytoplasm,smaller volume and looser arrangement.The leap tube and the lumen of the tube were obvious,the cells in the wall of the tube were arranged neatly,the capillary endothelium was not hyperplastic,the epithelial cells were flat,and did not protrude into the lumen.There is a small infiltration of lymphocytes around the local vessels.In the parotid gland tissue of rats with kidney deficiency syndrome group,there was reduced cytoplasm and loose arrangement of acinus cells,accompanied by punctate infiltration of lymphocytes.(8)Immunohistochemical results: Compared with the normal control group,the protein expressions of AQP1 and AQP5 in the parotid gland of the kidney deficiency constitution group and the kidney deficiency syndrome group had a tendency to decrease,and the average optical density value was decreased,with statistical significance(P <0.01).Compared with the kidney deficiency group,the expression of AQP5 in the kidney deficiency group was decreased,and the average optical density value was statistically significant(P < 0.05).(9)Western blot analysis of AQP1,AQP5,NF-κBp65 and NF-κBP-p65 in parotid gland: Compared with the normal control group,the expressions of AQP1 and AQP5 in renal deficiency group were decreased,while the expressions of NF-κBp65 and NF-κBP-p65 were increased,with statistical differences(P < 0.05 or P < 0.01).The expressions of AQP1 and AQP5 in the kidney deficiency group were significantly decreased,with statistical significance(P < 0.01).The expression of NF-κBp65 and NF-κBp-p65 were significantly increased,and the difference was statistically significant(P < 0.01).Compared with the kidney deficiency group,the expression of APQ1 was decreased and the expression of NF-κBp65 was increased in the kidney deficiency group,with statistically significant differences(P < 0.01).The expression of AQP5 tended to decrease,and the expression of NF-κBP-p65 tended to increase,but there was no statistical significance(P > 0.05)(10)Results of m RNA expression of NF-κBp65 protein in parotid gland: Compared with normal control group,the m RNA of NF-κBp65 protein in parotid gland of rats in kidney deficiency constitution group and kidney deficiency syndrome group was increased,but there was no statistical significance(P > 0.05).Conclusion: With “normal constitution-kidney empty physical-kidney deficiency syndrome” dynamic evolution process of the growth situation,the general condition of rats,parotid gland weight,body weight,viscera than saliva flow,saliva,osmotic pressure,TNF alpha of saliva,salivary gland tissue AQP1 protein,AQP5 protein and nf-kappa Bp65 protein,the nf-kappa indicators,such as Bp-p65 protein has changed,and with the state of the kidney is aggravating,the corresponding index of change is more obvious,this shows a rule that the secretion of saliva is closely associated with the rise and fall of renal function,and as the state of the kidney,The expression of aquaporin in parotid gland decreased gradually,which affected the secretion of saliva.The mechanism may be related to the regulation of the expression of aquaporin by TNF-α/NF-κB signaling pathway. |
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