防己黄芪汤治疗尿酸性肾病的初步机制研究文献综述

防己黄芪汤治疗尿酸性肾病的初步机制研究研究目的： 1．获得LRRK1基因调控肾脏尿酸代谢、进而调控UAN发生发展的可靠证据，丰富和完善UAN病理机制研究，为确立LRRK1基因相关通路作为UAN防治的新靶点提供更充分的科学依据。

2. 明确FHT是否通过LRRK1基因调控肾脏尿酸转运系统进而达到治疗UAN的目的，并更进一步明确LRRK1通过何种具体机制调控肾脏尿酸转运系统。

研究内容：动物整体实验利用OX建立HUA小鼠模型，通过对比分析正常组、HUA模型组、阳性药组和FHT给药组肾脏中LRRK1、PDZK1、NHERF1、EGFR及尿酸转运系统相关因子的表达，发现FHT降尿酸、促进尿酸排泄的可能分子靶点。

研究方案： 动物整体实验，建立HUA小鼠模型，证实FHT降尿酸、促进尿酸的排泄作用。

雄性SPF级昆明小鼠随机分为5组，每组12只，分别为正常对照组（生理盐水10 ml/kg）、模型对照组（OX 250 mg/kg）、FHT低、中、高剂量组、阳性药对照组（OX 250 mg/kg 别嘌呤醇10 mg/kg）。

给药周期为7 天。

在第5天给药1h后，每组分别将8只小鼠置入代谢笼，适应1天后，于给药第6天收集24 h尿液，2000g常温离心10 min，取上清液，测量尿液体积后按试剂盒方法测定尿液生化指标。

小鼠腹腔注射10%水合氯醛麻醉后采集血样，处死小鼠，取肾脏，检测血清生化指标，并采用实时定量PCR聚合酶链式反应法（qPCR）和蛋白质印迹法（WB）检测肾LRRK1、PDZK1、NHERF1、EGFR及尿酸转运系统相关因子的mRNA、蛋白及磷酸化蛋白的表达水平；免疫组化法（IHC）检测LRRK1在肾脏的分布。

主要成果形式：一篇毕业论文主要参考文献：[1] Hao, S, Zhang, C, Song, H, Natural Products Improving Hyperuricemia with Hepatorenal Dual Effects.. Evid Based Complement Alternat Med., 2016. 2016(10): p. 7390504.[2] Liu, YL, Pan, Y, Wang, X, et al., Betaine Reduces Serum Uric Acid Levels and Improves Kidney Function in Hyperuricemic Mice. Planta Med., 2014. 80(1): p. 39-47.[3] Ding, XQ, Pan, Y, Wang, X, et al., Wuling San ameliorates urate under-excretion and renal dysfunction in hyperuricemic mice. Chin J Nat Med., 2013. 11(3): p. 214-221.and renal dysfunction in hyperuricemic mice. Chin J Nat Med., 2013. 11(3): p. 214-221.[4] Sakiyama, M, Matsuo, H, Shimizu, S, et al., A common variant of leucine-rich repeat-containing 16A (LRRC16A) gene is associated with gout susceptibility. Hum Cell., 2014. 27(1): p. 1-4.[5] Vieira, AT, Galvao, I, Macia, LM, et al., Dietary fiber and the short-chain fatty acid acetate promote resolution of neutrophilic inflammation in a model of gout in mice. J Leukoc Biol., 2017. 101(1): p. 275-284.[6] Kocic, Gordana, Veljkovic, Andrej, Kocic, Hristina, et al., Depurinized milk downregulates rat thymus MyD88/Akt/p38 function, NF-kappa B-mediated inflammation, caspase-1 activity but not the endonuclease pathway: in vitro/in vivo study. Sci Rep., 2017. 7: p. 41971.[7] M, Luo, Sunil Yeruva, YJ, Liu, et al., IL-1beta;-Induced Downregulation of the Multifunctional PDZ Adaptor PDZK1 Is Attenuated by ERK Inhibition, RXRalpha;, or PPARalpha; Stimulation in Enterocytes. Front Physiol., 2017. 8: p. 61.[8] Mejia-Rangel, J, Cordova, E, Orozco, L, et al., NHERF1 regulates the progression of colorectal cancer through the interplay with VEGFR2 pathway. Oncotarget, 2017. 8(5): p. 7753-7765.[9] Mejia-Rangel, J, Cordova, E, Orozco, L, et al., Pro-adhesive phenotype of normal endothelial cells responding to metastatic breast cancer cell conditioned medium is linked to NF kappa B-mediated transcriptomic regulation. Int J Oncol., 2016. 49(5): p. 2173-2185.[10] Hanafusa, H, Matsumoto, K, LRRK1 regulates spindle orientation by phosphorylating CDK5RAP2. Cell Cycle., 2015. 14(21): p. 3349-3350.[11] Morimoto, K, Baba, Y, Shinohara, H, et al., LRRK1 is critical in the regulation of B-cell responses and CARMA1-dependent NF-kappa B activation. Sci Rep., 2016. 6.[12] Liu, N., L. Wang, T. Yang, et al., EGF Receptor Inhibition Alleviates Hyperuricemic Nephropathy. J Am Soc Nephrol., 2015. 26(11): p. 2716-2729.[13] Zeng, M, Chen, BF, Qing, YF, et al., Estrogen Receptor Beta Signaling Induces Autophagy And Downregulates Glut9 Expression. Nucleosides Nucleotides Nucleic Acids., 2014. 33(7): p. 455-465.[14] Ren, XY, Yuan, L, Shen, SJ, et al., c-Met and ER beta expression differences in basal-like and non-basal-like triple-negative breast cancer. Tumour Biol., 2016. 37(8): p. 11385-11395.[15] Kedashiro, S, Pastuhov, SI, Nishioka, T, et al., LRRK1-phosphorylated CLIP-170 regulates EGFR trafficking by recruiting p150(Glued) to microtubule plus ends. J Cell Sci., 2015. 128(2): p. 385-396.[16] Wang, R, Ma, CH, Zhou, F, et al., Siwu decoction attenuates oxonate-induced hyperuricemia and kidney inflammation in mice. Chin J Nat Med., 2016. 14(7): p. 499-507.[17] Yang, H, Gao, LH, Niu, YF, et al., Mangiferin Inhibits Renal Urate Reabsorption by Modulating Urate Transporters in Experimental Hyperuricemia. Biol Pharm Bull., 2015. 38(10): p. 1591-1598.[18] Wang, M, Zhao, J, Zhang, N, et al., Astilbin improves potassium oxonate-induced hyperuricemia and kidney injury through regulating oxidative stress and inflammation response in mice. Biomed Pharmacother., 2016. 83: p. 975-988.本课题工作进度：2018.3~2018.4：采用OX建立HUA小鼠模型，初步明确FHT通过干预LRRK1基因而调控肾脏尿酸转运系统，进而缓解UAN的作用机制和分子靶点。

2018.5：完成毕业论文。