高脂及劳倦伤脾大鼠有机阴离子转运肽oatp4a1 mRNA及蛋白表达

目的 观察有机阴离子转运肽oatp4a1在脾虚模型大鼠各组织中的表达。探讨其在水液湿浊转运中的作用机制。方法 选取6只SPF级大鼠制备劳倦伤脾模型，另选取12只大鼠随机分为空白对照组(空白组)、高脂饮食组(高脂组)，每组6只。劳倦伤脾模型组(劳倦组)单号日期采用特制束缚筒，每次3 h，双号日期令大鼠在冷水［(10±1)℃］中游泳7 min，连续2周。劳倦组、空白组常规标准大鼠颗粒饲料喂养12周，高脂组大鼠以高脂饲料喂养12周，均自由饮食饮水。采用RT-PCR和Western blot法检测肺、脾、肝、肾、胃、小肠、大肠组织oatp4a1 mRNA、蛋白表达。结果 劳倦组大鼠扎堆懒动，烂便，毛色粗糙, 饮食饮水量减少，体重下降(P<0.05)，与临床脾虚证表现基本相符, 提示劳倦伤脾动物模型造模全部成功。高脂组大鼠第9周之后开始出现食欲不振，精神萎靡，嗜卧懒动，毛发疏松粗糙，晦暗无光泽等脾虚症状，形成痰湿证大鼠。与空白组比较，高脂组第9周平均体重升高(P<0.05)。oatp4a1 mRNA在肺、脾、肝、肾、胃、小肠、大肠7种脏器中都有表达；各组各组织oatp4a1 mRNA表达量比较，差异均无统计学意义(P>0.05)；高脂组大鼠肺、肾oatp4a1 mRNA表达量均高于肝(P<0.05)。结论 oatp4a1可能是高脂伤脾大鼠机体参与湿浊转运的物质基础之一，其中肺、肾、大肠在湿浊转运中可能起着重要作用。

Organic Anion Transporting Polypeptide(oatp4a1) mRNA and Protein Expressions in High Fat and Over-fatigue Impairing Pi Rats

Objective To study organic anion transporting polypeptide(OATP) superfamily member 4a1(oatp4a1) mRNA expression in the Pi deficiency model rats,thus exploring its mechanism for transporting and transforming the dampness.Methods Six SD rats of SPF grade were used to prepare over-fatigue impairing Pi model.Another 12 SD rats were randomly divided into the blank control group and the high fat diets group,6 in each.The special binding tube was used for the over-fatigue impairing Pi model group on the odd day,3 h each time.Then the rats were forced to swim in the cold water(10 ℃±1 ℃) for 7 min on the even day,for 2 successive weeks.Rats in the model group and the blank control group were granulatedly fed for 12 weeks,while rats in the high fat group were fed with high fat diet for 12 weeks.All rats were free to take food and drink water.The mRNA and protein expressions of oatp4a1 were detected in the Fei,Pi,Gan,Shen,Wei,Xiaochang,and Dachang using Real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)and Western blot.ResultsRats in the model group were idled together with lousy defecate and coarse skin.They ate and drank less,and lost body weight(P<0.05).They were consistent with clinical manifestations of Pi deficiency syndrome,indicating that the over-fatigue impairing Pi animal model was successfully established.Rats in the high fat group started to have poor appetite and languish spirit,move lazily and addict to sleep,have coarse,dark,and colorless hair 9 weeks later,indicating phlegm dampness syndrome.Compared with the blank control group,the average body weight increased in the high fat group at the 9th week(P<0.05).The oatp4a1 mRNA expressed in the Fei,Pi,Gan,Shen,Wei,Xiaochang,and Dachang.There was no statistical difference in the oatp4a1 mRNA expression among all tissues(P>0.05).The oatp4a1 mRNA expressions were higher in the Fei and Shen of the high fat group than in the Gan(P<0.05).Conclusions oatp4a1 might be one of the basic substances in the transportation and transformation of phlegm dampness.Of them,Fei,Shen,and Dachang might play important roles in the transportation and transformation of phlegm dampness.