| Objective:By using solvent extraction system Yishen Huayu Recipe screening,four different polar fractions obtained (Yishen Huayu Recipe petroleum ether group,ethyl acetate extract Yishen Huayu Recipe group of kidney Founder butanol Fractionstasis, Yishen Huayu Recipe group parts water), dynamic model to observe the ratkidney with unilateral ureteral obstruction (UUO) function, renal morphologicalchanges in renal tissue Col-I,Col-IV,Hyp Pathological changes in the content andprocess of α-SMA, changes in the expression of E-cadherin, and explore various partsbetween its anti-renal interstitial fibrosis possible mechanism.Methods:Yishen Huayu Recipe continuously with80%ethanol reflux extraction,evaporated to a thick paste no alcohol taste, followed by petroleum ether, ethyl acetate,n-butanol, respectively, to obtain petroleum ether, ethyl acetate extract, n-butanolparts, parts of four different polar water sites. While the level of SPF SD rats wererandomly divided into147sham operation group, model group, Yishen Huayu Recipepetroleum ether group, Yishen Huayu Recipe ethyl acetate extract group, kidneycirculation butanol fraction Founder group, kidney water site Huayu Recipe group andvalsartan group, each21, with unilateral ureteral obstruction ligation rat model UUO,only blunt dissection without ligation of the left ureter in the sham group, I UUOgroup are the same.All animals were fed at the beginning of the next day modelingsham group and model group with an equal volume of saline, I drenched kidney wereHuayu Recipe petroleum ether, ethyl acetate, n-butanol, water-saturated4differentparts of the polarity of the drug (equivalent to Yishen Huayu Recipe16g/kg/d),valsartan group fed valsartan suspension13.3mg/kg.d, according2ml/100g/d gavage.All the animals were free access to water. Weighed every three days to adjust the dose.Rats in each group, respectively, after the first3,7,14d, each group were randomlyselected six, abdominal aortic blood detection of renal function; HE and Masson staining were used in light microscopy obstructed kidney pathological changesalkaline hydrolysis method to measure the content of Hyp, ELISA method to measureCol-I,Col-IV levels, detected by immunohistochemical staining α-SMA expression inrenal tissue and E-cadherin, and jointly evaluate the degree of fibrosis in renal injuryin order to improve as primary efficacy, renal function improved as a secondary effect,the site has the best access to effective screening pharmacological activity.Results:(1) The impact of the rats at different time points of renal functionSham group rats serum BUN, Scr and CysC each time point did not changesignificantly; UUO model group, serum BUN, Scr and CysC with time graduallyincreased; serum remaining after each drug group intervention BUN, Scr and CysC ateach time point was significantly lower than the model group; among the various partsof the treatment of Yishen Huayu Recipe group, n-butanol most obvious part of thegroup, and no significant difference between the two valsartan group (P>0.05).HuayuRecipe affect kidney tissue UUO model of renal pathological changes(2)Effects of pathological changes in kidney tissue of rats.HE, Masson staining: sham-operated rats at each time point no significant changein the organizational structure of the kidney; kidney tissue lesions UUO model group,the most serious, and gradually increased with time, mainly for the small tubeexpansion, inflammation, edema etc., fibrosis gradually increased with time; ratkidney tissue lesions each drug intervention group at each time point was significantlylighter than the model group, which in various parts of the treatment of Yishen HuayuRecipe, n-butanol and lightest parts of the lesion group.(3)Changes at different time points in the renal tissue of rats Hyp contentRenal tissue of rats in the sham group at each time point Hyp content did notchange significantly; rat kidney tissue Hyp UUO model group were significantlyincreased, and gradually increased with time; rest of the drug intervention in renaltissue of rats Hyp was significantly lower than the untreated group, in which each partof the treatment of kidney Huayu Recipe group, n-butanol most obvious part of thegroup, and no significant difference between the two valsartan group (P>0.05).(4)Col-I, changes in renal tissue at different time points in each group of rats Col-IVcontentRenal tissue of rats in the sham group Col-I,Col-IV showed no significantchange; kidney tissue UUO model group Col-I,Col-IVwas significantly increased, and increased with time; rest of the drug intervention group were renal tissu eCol-I,Col-IVwas significantly lower than the untreated group, in which each part of the treatment of Yishen Huayu Recipe group, n-butanol most obvious part of the group, and no significant difference between the two valsartan group(P>0.05).(5)Different time points in the renal tissue of rats α-SMA, E-cadherin expressionchange impactsRenal tissue of rats in the sham group only see the middle arterial smooth musclecells have less α-SMA expression, and renal tubular and interstitial cells did notexpress, and no significant change at each time point; UUO model rats3days afterkidney a large brown cells expressing α-SMA-positive interstitial emerged, and withthe extension of time was significantly increased; each drug intervention in renaltissues of rats at each time point α-SMA in significantly less than the model group,which kidney Huayu Recipe various parts of the treatment group, n-butanol mostobvious part of the group, and no significant difference between the two valsartangroup (P>0.05), E-cadherin is the opposite.Renal tissue of rats in the sham group, mainly in the cell membrane E-cadherinexpression, brownish yellow, and each time no significant change; UUO model ratsafter3days, a significant reduction in E-cadherin expression in renal tubularepithelial cells, and with prolonged obstruction continue to decrease; each drugintervention renal tissue expression of E-cadherin positive at each time point wassignificantly more than the untreated group, in which each part of the treatment ofYishen Huayu Recipe group, n-butanol is most effective part of the group obviously,both the valsartan group and no significant difference (P>0.05), α-SMA is theopposite.Conclusion：1. Using solvent extraction system, successful kidney Huayu Recipepetroleum ether, ethyl acetate extract kidney Huayu Fang, kidney and kidneycirculation Founder butanol parts water circulation party site. Four polar sites, thekidney circulation Founder butanol fraction group can significantly reduce damageUUO rat kidney tissue pathology, and decreased serum BUN UUO rats, Scr, CysClevels, lower renal tissue Hyp content, its role valsartan considerable effect, while theremaining three smaller improvement polar fractions, confirming kidney circulationFounder butanol fraction group has better delay between renal interstitial fibrosis.2.kidney circulation Founder butanol fraction group can significantly reducethe renal tissue Col-I, Col-IV levels, inhibit the expression of renal tissue α-S MA, and regulate the expression of E-cadherin, indicating that by reducing ECM accumulation, reduce the degree of renal interstitial fibrosis, so as to achieve the antagonistic renal interstitial fibrosis.3. Kidney circulation Founder alcohol group may be an effective part of the site Huayu Recipe antagonistic kidney renal interstitial fibrosis. |
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