Based On β₃-AR To Investigate The Reducing Urination And Nourishing Kidney-yang Mechanism Of SQW

According to TCM's theory, Kidney is the origin of congenital constitution and kidney deficiency is one of the primarily causes of aging. If Kidney-Qi is adequate, the bladder can be much warmed, right astringent, opening and closing of a degree, in order to maintain normal metabolism of liquid water. If congenital deficiency or illness disorders, weak ferrite cause kidney deficiency, Xiayuan virtual cold, loss of bladder opening and closing effect, bladder dysfunction of gasification, which causes polyuria.The formula of Suoquanwan (SQW) is recorded in a Chinese medicine book named Furenliangfan, which collected hundreds of formulas of Chinese medicine combinations. SQW consists of three herb medicinesc Radix Linderase, Radix dioscorteae, and alpinia oxyphylla Miq. The function of SQW is to warm kidney, stop polyuria and hold excessive urination. In clinical practice, SQW is used for the treatment of urinary system diseases alone or be used in combination with other drugs, which is Polyuria and excessive urination due to Yang-deficiency of Kidney, such as enuresis, diabetes insipidus, noeturnalenuresis, neurogenic urinary frequen, chronic glomerulonephritis, nephrotic syndrome, urethral syndrome.There is a higher incidence of bladder dysfunction in the elderly, in addition to such a clear cause as BOO, weak pelvic floor supporting tissues and nerve disorders, but can still be complicated by clinical or unexplained single urinary frequency, urgency, and incontinence and so on, for which neurotransmitters have poor drug efficacy, indicating that there are aging-related changes in detrusor own characteristics. Relaxation mechanism of the bladder, inhibition of cholinergic receptor (M receptor) and exciting ofβ-adrenergic receptor (β-AR) is the most major. However, the number of M receptor and affinity with increasing age did not change significantly, but reactivity ofβ-AR in the detrusor cell decreased, AC activity and cAMP levels decreased correspondingly.β-AR subtypes are involved in the regulation of detrusor relaxation, but detrusor relaxation mediated mainly by theβ3-AR.β3-AR and M receptor antagonistic effect can be maintained bladder compliance and stability. Changes of its structure and function are closely related to urine storage and bladder dysfunction in the elderly.Previous studies of this subject have shown that SQW has a significant anti-diuretic effect on rats with water loading test and mice with a diuretic drug. After treatment, Na+, Cl- ion excretion was significantly reduced, while K+ excretion increased, which is similar to effects of aldosterone. Therefore, the mechanism of synthesis and secretion of aldosterone was investigated in kidney-deficiency rats with polyuria after administration SQW. While the study were processed from improving the HPA axis (hypothalamus-pituitary-adrenal axis) act and micturition central neurotransmitter and its receptor, gene and protein expression side, which also based on the "kidney" and the relevance of the neuroendocrine system. In addition, aquaporin 2 (AQP2) expressions and signal transduction pathway of kidney-deficiency rats with polyuria model after administration SQW also were studied. The theory of "warming kidney and stopping ployuria" were investgated respectively from neuroendocrine, renal tubular reabsorption and secretion of various aspects of starting depth at the molecular level, which laid the foundation of research about the subject.In view of this, the subject based on the neuroendocrine system the early renal tubular function studies, researched the old-Kidney deficiency and ployuria-Bladder detrusor-nerve receptors together, intended to regulate the diastolic function of the bladder detrusor neural receptorβ3-AR and its signal transduction research as a starting point, combined with endocrinology and immunological indicators, researched molecular pharmacology mechanism "nourishing kidney and stopping urination".SQW can reduced the urination volume in the kidney-deficiency rats with polyuria model, which was investigated by studying preliminary urine and urinary Na+, K+, Cl- concentration of natural aging rats in this experiment. Regulation of SQW on kidney-deficiency rats with polyuria model was observed from the endocrine perspective through research the effects of plasma levels of ADH and ALD in rats. Effects of SQW on the content of Cort in serum was observed by ELISA and SWQ play a effect of warming kidney was observed by influence of SQW on thymus, spleen and adrenal weight. Effects of SQW on expression ofβ3-ARmRNA was detected by using fluorescence quantitative PCR and in situ hybridization method in the bladder detrusor of kidney-deficiency rats with polyuria model. Effects of SQW on the content of AC, cAMP and PKA was tested by using ELISA in the bladder detrusor of kidney-deficiency rats with polyuria model. Effects of SQW on the PKA protein expression was detected by western blotting in the bladder detrusor of kidney-deficiency rats with polyuria model. The mechanism of warming kidney and stopping ployuria" theory about SQW was evaluated from the perspective of molecular pharmacology.It is a good and ideal kidney of Yang-deficiency and polyuria model. SQW can significantly reduce the urination volume in kidney-yang deficiency rats with polyuria. SQW can reduce Na+, Cl- emission and increased K+ excretion in kidney-yang deficiency rats with polyuria. SQW can increase the content of ALD and ADH in blood of kidney of Yang-deficiency and polyuria mode. SQW can increase significantly the spleen, thymus, adrenal coefficient and improved kidney spleen and thymus, adrenal atrophy state in kidney-yang deficiency rats with polyuria. SQW can increase the content of Cort in blood of kidney-yang deficiency rats with polyuria. SQW can elevate significantly the expression ofβ3-ARmRNA in bladder detrusor of kidney-yang deficiency rats with polyuria. SQW can also increase the content of AC, cAMP and PKA in bladder detrusor of kidney-yang deficiency rats with polyuria. SQW can elevate significantly the expression of PKA protein in bladder detrusor of kidney-yang deficiency rats with polyuria.β3-AR of adjusting the bladder detrusor relaxation neural receptor and its signal transduction research were selected as a starting point, which deeply interpreted and clarified the basis of molecular pharmacology of SQW treatment diseases of Kidney-deficiency and polyuria, fully illustrated Chinese herbal compound multi-pass, multi-target, the overall adjustment of the treatment characteristics, provided new ideas and methods for the classical formula of SQW and modern research of "warming kidney and stopping ployuria" theory.