Dai Sheng-Jie, Zhang Qiao-Yun, Lan Qing, Chen Yong, Zhang You-Zhi, Huang Qi
School of Pharmacy, Hubei University of Science and Technology Xianning 437100, China.
Department of Nephrology, Xianning Central Hospital of Hubei Province Xianning 437100, China.
Zhongguo Zhong Yao Za Zhi. 2022 Mar;47(6):1611-1617. doi: 10.19540/j.cnki.cjcmm.20211122.401.
This study aimed to investigate the effects of geniposide(GP) on the expression of prokineticin(PK2) and prokineticin receptor 1(PKR1) in db/db mice with diabetic nephropathy(DN), so as to explore how the PK2 signaling pathway participated in the pathological changes of DN and whether GP exerted the therapeutic effect through this signaling pathway. Male mice were randomly divided into four groups, namely db/m, db/db, db/db+GP, and db/m+GP groups, with five in each group. The mice in the db/db+GP and db/m+GP groups were gavaged with 150 mg·kg~(-1) GP for eight successive weeks. Afterwards, all the mice were sacrificed and the renal tissues were embedded. The morphological changes in glomerulus and renal tubules were observed by Masson and PAS staining. The expression levels of PK2, PKR1, and Wilm's Tumor Protein 1(WT_1) in podocytes were detected by immunohistochemistry, and the protein expression levels of PK2 and PKR1 in mouse kidney by Western blot. The morphological results showed serious glomerular and tubular fibrosis(Masson), high glomerular and tubular injury score(PAS), increased glomerular mesangial matrix, thickened basement membrane, exfoliated brush border of renal tubules, decreased WT_1 in glomerular podocytes, and massive loss of podocytes in the db/db group. After administration with GP, the glomerular and tubular fibrosis was alleviated, accompanied by improved glomerular basement membrane and renal tubule brush edge, and up-regulated WT_1. As revealed by further protein detection, in the db/db group, the expression levels of PK2 and PKR1 and p-Akt/Akt ratio declined, whereas the ratio of Bax/Bcl-2 rose. Ho-wever, PKR2 and p-ERK/ERK ratio did not change significantly. After administration with GP, the PK2 and PKR1 expression was elevated, and p-Akt/Akt ratio was increased. There was no obvious change in PKR2, Bax/Bcl-2 ratio, or p-ERK/ERK ratio. All these have demonstrated that GP improves the renal damage in DN mice, and PK2/PKR1 signaling pathway may be involved in such protection, which has provided reference for clinical treatment of DN with GP.
本研究旨在探讨栀子苷（GP）对糖尿病肾病（DN）db/db小鼠中前动力蛋白2（PK2）及前动力蛋白受体1（PKR1）表达的影响，以探究PK2信号通路如何参与DN的病理变化以及GP是否通过该信号通路发挥治疗作用。雄性小鼠随机分为四组，即db/m、db/db、db/db+GP和db/m+GP组，每组五只。db/db+GP组和db/m+GP组小鼠连续八周灌胃150 mg·kg⁻¹ GP。之后，处死所有小鼠并将肾组织包埋。通过Masson和PAS染色观察肾小球和肾小管的形态变化。采用免疫组化法检测足细胞中PK2、PKR1及威尔姆斯瘤蛋白1（WT_1）的表达水平，并用蛋白质印迹法检测小鼠肾脏中PK2和PKR1的蛋白表达水平。形态学结果显示，db/db组存在严重肾小球及肾小管纤维化（Masson染色）、肾小球及肾小管损伤评分高（PAS染色）、肾小球系膜基质增多、基底膜增厚、肾小管刷状缘脱落、肾小球足细胞WT_1减少及足细胞大量丢失。给予GP后，肾小球及肾小管纤维化减轻，伴有肾小球基底膜及肾小管刷状缘改善，WT_1上调。进一步的蛋白质检测结果显示，db/db组中PK2和PKR1的表达水平及p-Akt/Akt比值下降，而Bax/Bcl-2比值升高。然而，PKR2及p-ERK/ERK比值无明显变化。给予GP后，PK2和PKR1表达升高，p-Akt/Akt比值增加。PKR2、Bax/Bcl-2比值及p-ERK/ERK比值无明显变化。所有这些均表明，GP可改善DN小鼠的肾损伤，PK2/PKR1信号通路可能参与了这种保护作用，这为临床应用GP治疗DN提供了参考。
