| ObjectiveEdema is one of the main clinical manifestations of nephrotic syndrome. Deficiency of spleen yang kidney yang is the most common pathogenesis of edema in nephrotic syndrome. Both warming yang to resolve fluid retention and warming yang to promote diuresis are often used in treating edema. From the perspective of traditional Chinese medicine, both of two methods belong to warming methods and are applicable to regulate watermetabolism imbalance syndromes caused by deficiency of yang. Warming yang to resolve fluid retention is to regulate water metabolism imbalance syndromes caused by deficiency of heart and spleen yang which located at upper-jiao and middle-jiao, but warming yang to promote diuresis is to regulate water metabolism imbalance syndromes caused by deficiency of kidney yang which located at lower-jiao. But from the view of modern medical research, differences between two kinds of treatmentmechanism is still not clear. Acoording to water metabolism theory of traditional Chinese medicine and modern medicine understanding of aquaporins (AQP), we infer that these two methods regulate water metabolism by adjusting aquaporins, and hypothesize that "warming yang to resolve fluid retention regulate water metabolism by adjusting aquaporins in digestive tract mainly, and warming yang to promote diuresis regulate water metabolism by adjusting aquaporins in kidney mainly". This study tried to clarify intrinsic basis of the effect difference between these two methods by expression of aquaporins in digestive tract and kidney, AVP and cAMP which are main adjustment factors of aquaporins. First, we studied expression of aquaporins and level of AVP and cAMP of edema patients with primary nephrotic syndrome and non-edema patients with chronic nephritis. In order to reveal internal basis of clinical effect differences between these two methods, we compared difference of expression of aquaporins and level of AVP and cAMP of adriamycin nephropathy rats that were treated by warming yang to resolve fluid retention and warming yang to promote diuresis respectively.MethodsIn the clinical research, studied cases were hospitalized patients who were from department of nephrology of Guangdong Provincal Hospital of Traditional Chinese Medicine during2013June to2014February. Studied cases were dividied into two groups, edema with primary nephrotic syndrome groups and non-edema with chronic nephritis groups. Control cases were health residents from Guangzhou community who were surveyed in2012May. All the studied cases leaved urine, blood samples and kidney tissue samples, and control cases leaved urine and blood samples. We tested serum AVP and cAMP and urine AQP2by Elisa, and then tested expression of AQP1in kidney by immunohistochemistry.In the experimental study,32male SD rats were divided into4groups: blank control group, pathologic control group, warming yang to resolve fluid retention group, warming yang to promote diuresis group. Pathologic control group, warming yang to resolve fluid retention group and warming yang to promote diuresis group were treated by adriamycin induced nephropathy model. Then Lingguizhugan Decoction, Zhenwu Decoction were used as the carrier of warming yang to resolve fluid retention and warming yang to promote diuresis respectively.All the rats wer treated for4weeks after modeling success. At the end of the experiment, we tested serum AVP, cAMP level and urine AQP2level by Elisa, and tested expression of aquaporins in stomach, kidney and ileum by immunohistochemistry and weston-blot, and detected relative expression of mRNA of aquaporins in stomach, kidney and ileum by real time PCR.ResultIn the clinical study,15edema patients with primary nephrotic syndrome and15non-edema patients with chronic nephritis,10healthy people were included. Compared with healthy control group, urine AQP2of edema group of primary nephrotic syndrome increased significantly (P<0.01), and serum cAMP decreased significantly (P<0.01).And urine AQP2of non-edema group of chronic nephritis also increased (P<0.05), and serum cAMP decreased (P<0.05). But there was no statistically significant of urine AQP2and serum cAMP between of these two groups. There was no statistically significant difference of serum AVP between these three groups. The expression of AQP1which mainly expressed in basolateral membrane of proximal tubular epithelial cells in renal cortex of edema group of primary nephrotic syndrome increased(P<0.05).In the experimental research, there were no statistically significant differences of urinary volume, serum AVP and cAMP between warming yang to resolve fluid retention group and warming yang to promote diuresis group. Compared to blank control group,24hours urine AQP2of warming yang to promote diuresis group increased significantly (P<0.01) and warming yang to resolve fluid retention group increased (P<0.05), but there was no statistically significant difference between them. In kidney of Adriamycin nephropathy rat model, Immunohistochemistry indicated that warming yang to promote diuresis inhibited the high expression of AQP2(P<0.05), and warming yang to resolve fluid retention inhibited the high expression of AQP1in ileum grands (P<0.05). Westonblot indicated that warming yang to promote diuresis inhibited the high expression of AQP4in kidney of Adriamycin nephropathy rat model (P<0.05). Real time PCR indicated that there was no significantly difference between warming yang to resolve fluid retention and warming yang to promote diuresis on regulating AQPl/2/3/4mRNA of kidney and AQP1/3/4mRNA of stomach and ileum of Adriamycin nephropathy rat model. ConclusionUrine AQP2and expression of AQP1in renal cortex tissue increased in patients with nephrotic syndrome. It provided clinical basis for the further study of mechanism of warming yang to resolve fluid retention and warming yang to promote diuresis that act on edema of nephrotic syndrome. Both warming yang to resolve fluid retention and warming yang to promote diuresis regulared expression of aquaporins in kidney. But warming yang to resolve fluid retention regulared expression of AQP1in digestive tract mainly. Warming yang to promote diuresis regulared expression of AQP2and AQP4in kidney mainly. And it is generally consistent with the hypothesis of "warming yang to resolve fluid retention regulate water metabolism by adjusting aquaporins in digestive tract mainly, and warming yang to promote diuresis regulate water metabolism by adjusting aquaporins in kidney mainly"... |
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