茯苓对阴虚水肿模型大鼠及其热象的影响

Effect of Poria cocos on yin deficiency edema rats and heat syndrome

目的观察茯苓对阴虚水肿模型大鼠的影响,研究茯苓利水作用特点及机制,并评价茯苓对阴虚水肿所表现出热象的影响。方法尾iv阿霉素联合ig甲状腺片复制阴虚水肿模型大鼠,给予茯苓水煎液或六味地黄丸干预4周,观察大鼠水肿情况和一般生长状态,检测24 h尿量和尿蛋白,测量肛温,进行冷热板实验计算热板停留时间比;检测血清总蛋白(total cholesterol,TP)、白蛋白(albumin,Alb)、尿素氮(blood urea nitrogen,BUN)、肌酐(creatinine,Cr)、环磷酸腺苷(cyclic adenosine monophosphate,c AMP)、环磷酸鸟苷(cyclic guanosine monophosphate,c GMP)水平;测定肾组织超氧化物歧化酶(superoxide dismutase,SOD)活性及活性氧(reactive oxygen species,ROS)、丙二醛(malondialdehyde,MDA)水平;苏木素-伊红(HE)染色观察肾组织病理变化;Western blotting检测肾脏组织Klotho和棕色脂肪组织中解偶联蛋白1(uncoupling protein 1,UCP1)、过氧化物酶体增殖物激活受体γ辅助激活因子-1α(peroxisome proliferator activated receptorγcoactivator-1α,PGC-1α)蛋白表达;qRT-PCR检测肾脏组织Klotho和棕色脂肪组织中UCP1、PGC-1αm RNA表达。结果茯苓可使模型大鼠水肿情况好转,显著增加24 h尿量(P<0.05),减小24 h尿蛋白(P<0.05);显著升高血清中TP、Alb(P<0.05),降低BUN、Cr(P<0.05);显著升高肾脏SOD活性(P<0.05),降低MDA、ROS水平(P<0.05);对大鼠肛温、热板停留时间比、血清中c AMP、c GMP水平及c AMP/cGMP值无明显影响;显著上调肾脏Klotho m RNA及蛋白表达(P<0.05),对棕色脂肪组织UCP1、PGC-1αmRNA及蛋白表达无明显影响;光镜下可见肾小球系膜增生改善,系膜基质减少。结论在阴虚水肿模型大鼠上,茯苓通过抗氧化应激改善阿霉素诱导的肾损伤而表现出利水作用,但茯苓对病证的热象无明显影响。

Objective To observe the effect of Poria cocos on yin deficiency edema model rats,study the characteristics and mechanism of P.cocos diuresis,and evaluate the effect of P.cocos on yin deficiency edema.Methods Adriamycin was injected into tail vein and thyroxine tablets were given by gavage to replicate yin deficiency edema model rats.P.cocos decoction or Liuwei Dihuang Pills(六味地黄丸)was given by gavage for four weeks.The edema and general growth state of rats were observed,urine volume and proteinuria in 24 h were detected,anal temperature was measured,and residence time ratio of hot plate was calculated by cold-hot plate experiment.Serum total protein(TP),albumin(Alb),blood urea nitrogen(BUN),creatinine(Cr),cyclic adenosine monophosphate(cAMP)and cyclic guanosine monophosphate(cGMP)were detected.The activity of superoxide dismutase(SOD)and levels of reactive oxygen species(ROS)and malondialdehyde(MDA)in kidney tissue were measured.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of kidney tissue.Western blotting was used to detect Klotho protein expression in kidney and uncoupling protein 1(UCP1),peroxisome proliferator activated receptorγcoactivator-1α(PGC-1α)protein expressions in brown adipose tissue.qRT-PCR was used to detect Klotho mRNA expression in kidney and UCP1,PGC-1αmRNA expressions in brown adipose tissue.Results P.cocos improved the edema of model rats,significantly increased 24 h urine volume(P<0.05),decreased 24 h urine protein(P<0.05).Serum TP and ALB were significantly increased(P<0.05),BUN and Cr were decreased(P<0.05).Activity of SOD in kidney was significantly increased(P<0.05),levels of MDA and ROS were decreased(P<0.05);There was no significant effect on anal temperature,hot plate retention time ratio,serum cAMP and cGMP levels and cAMP/cGMP in rats.Klotho mRNA and protein expressions in kidney were significantly increased(P<0.05),but UCP1 and PGC-1αmRNA and protein expressions in brown adipose tissue were not significantly affected.Microscopically,glomerular mesangial hyperplasia was improved and mesangial matrix was decreased.Conclusion P.cocos can improve the kidney injury induced by doxorubicin through antioxidant stress in yin deficiency edema model rats,but it has no obvious effect on the thermal image of disease.