RNA-Seq analysis of kidney from female 5/6Nx B6 mice and female sham B6 mice. Mus musculus

Aim We have reported that D-Ser and D-Ala are protective against renal injury in acute kidney injury model mice. We generated 5/6-Nx-induced chronic kidney disease model mice with more severe renal injury and evaluated the protective effects of D-Ser and D-Ala. Methods Mice WT C57BL/6 mice were purchased from CLEA Japan (Osaka, Japan). All mice were housed and bred at Kanazawa University. D-Ser and D-Ala (Wako, Tokyo, Japan) was administrated orally at a concentration of 20 mM in distilled water. Mice were allowed free access to D-Ala dissolved water. 5/6-Nx Induction Progressive renal failure was induced via 5/6-Nx in a two-step procedure using 6-8 weeks mice. For initial 2/3 nephrectomy, the left kidney was exposed via flank incision. Clamp the renal arteries and veins, Upper and lower kidney poles were cut using bipolar. Furthermore, liquid thrombin (MOCHIDA PHARMACEUTICAL CO., LTD. Tokyo, Japan) was attached to the cut surface to stop bleeding. Right kidney nephrectomy was carried out after two weeks of recovery. Removal of the second kidney represents day 0. Sham surgery was also performed as a two-step procedure in which the respective kidney was exposed and then repositioned. The kidney samples enrolled in this study were treated with D-Ser and D-Ala and non-treated mice.

研究目的：此前我们已有报道称，D-丝氨酸（D-Ser）与D-丙氨酸（D-Ala）对急性肾损伤模型小鼠的肾损伤具有保护作用。本研究构建了肾损伤更为严重的5/6肾切除（5/6-Nx）诱导的慢性肾病模型小鼠，并以此评估二者的肾脏保护效应。
实验方法：野生型C57BL/6小鼠购自日本CLEA公司（日本大阪），所有小鼠均在金泽大学饲养繁育。D-丝氨酸与D-丙氨酸（购自日本和光纯药工业株式会社，日本东京）以20 mM浓度溶于蒸馏水，经口给药；小鼠可自由饮用溶解有D-丙氨酸的水溶液。
5/6肾切除模型构建：采用两步手术法对6~8周龄小鼠诱导进行性肾衰竭。首次手术行左侧2/3肾切除术：经侧腹切口暴露左侧肾脏，夹闭肾动静脉后，使用双极器械切除肾脏上下极，随后在创面涂抹液态凝血酶（购自日本持田制药株式会社，日本东京）以止血。术后恢复两周后行右侧肾脏切除术，右侧肾脏切除当日记为实验第0天。假手术组采用相同两步操作流程，仅暴露对应侧肾脏后即复位，不进行肾组织切除。本研究纳入的肾脏样本分别来自经D-丝氨酸、D-丙氨酸处理的小鼠以及未处理的对照组小鼠。