杞黄地黄丸中所含的黄耆和石榴籽, 源自于《小儿疾病治疗大全》中记载的 "六味地黄丸". 在传统上，它们被广泛应用于滋阴补肾的功效。 我们先前的研究表明杞黄地黄丸可保护腺嘌呤致慢性肾病 (CKD) 大鼠免受损害。然而，至今对于杞黄地黄丸影响CKD大鼠肾间质纤维化的机制仍没有明确的解释。本研究的目标是探索杞黄地黄丸对CKD大鼠肾间质纤维化的改善作用及潜在机制。通过腺嘌呤诱导大鼠CKD, 对两周后进行血液、尿液与肾组织的生化分析。通过大鼠的体重和肾指数变化观察大鼠的生理状态。通过酶联免疫吸附试验 (ELISA) 检测前炎症细胞因子，用苏木精伊红 (H&E) 和马松三色染色法评估肾组织的损伤和纤维化程度。为了确定与纤维化信号通路相关的指标和蛋白质水平，我们采用了实时聚合酶链式反应 (Rt-PCR)、西方印迹 (WB) 和免疫荧光技术。肾间质纤维化导致细胞功能受损，血尿素氮 (BUN)、尿蛋白 (UP)、白细胞介素-1β (IL-1β)、白细胞介素-6 (IL-6)和肿瘤坏死因子 alpha (TNF-α) 的水平上升，同时上调了胶原蛋白Ⅰ (COL-1)、纤维连蛋白 (FN)、α-平滑肌肌动蛋白 (a-SMA)、转化生长因子-β1 (TGF-β1)、c-Jun N-末端激酶 (JNK)、p38和细胞外调节蛋白激酶 (ERK)，下调了E-钙黏蛋白的表达。杞黄地黄丸显著改善了CKD大鼠的肾功能，表现为BUN、UP和肾指数的下降。此外，与肾组织形态学变化相符，经杞黄地黄丸干预后，CKD大鼠的肾间质纤维化显著改善，主要表现为COL-1、FN和a-SMA阳性表达的减少，以及E-钙黏蛋白水平的提高。同时，杞黄地黄丸减少了腺嘌呤诱导的CKD大鼠中的典型前炎症细胞因子IL-1β、IL-6和TNF-α。此外，杞黄地黄丸还下调了TGF-β1、JNK、p38和ERK。综上所述，杞黄地黄丸可能减少了腺嘌呤诱导的CKD大鼠中的炎症，改善了细胞外基质 (ECM) 成分的沉积，并减少了上皮间质转化标记蛋白的水平。此外，杞黄地黄丸的干预显著减少了炎症细胞因子的释放，抑制了TGF-β1/MAPK信号通路。根据结果，杞黄地黄丸可能对腺嘌呤诱导的肾纤维化有治疗作用。版权所有 © 2023. 由Elsevier B.V.出版。

Herb couple Rehmannia glutinosa Libosch and Cornus officinalis Sieb (RC), originated from "Liuwei Dihuang Pill" which recorded in Key to Therapeutics of Children's Diseases. Traditionally, they have been used widely for their ability to nourish yin and energize the kidneys. Our previous study indicated that the RC could protect against adenine induced Chronic kidney disease (CKD) rats. Nevertheless, there is still no clear explanation of the mechanisms by which RC affects renal interstitial fibrosis in CKD rats.Current Work aims to explore the amelioration and potential mechanism of RC on renal interstitial fibrosis in CKD rats.CKD rats were induced by adenine. Two weeks after administration, blood, urine, and kidney tissue were collected for biochemical analysis. Observing the physiological state of rats through the changes of rat body weight and renal index. The pro-inflammatory cytokines were measured by enzyme linked immunosorbent assay (ELISA), while renal tissue damage and fibrosis were assessed with Hematoxylin-eosin staining (H&E) and Masson's trichrome staining. In order to determine the levels of indicators and proteins associated with fibrosis signaling pathways, real time PCR (Rt-PCR), Western blot (WB), and immunofluorescence were employed.The renal interstitial fibrosis led to impaired cellular functions with increased the levels of Blood Urea Nitrogen (BUN), Urine protein (UP), Interleukin-1β (IL-1β), Interleukin-6 (IL-6), and Tumor Necrosis Factor alpha (TNF-α). and simultaneous up-regulated collagenⅠ(COL-1), fibronection (FN), α-smooth muscle actin (a-SMA), transforming growth factor-β1 (TGF-β1), c-Jun N-terminal kinase (JNK), p38 and extracellular regulated protein kinases (ERK), down-regulated the expression of the E-cadherin proteins. RC notably improved renal dysfunction in CKD rats as indicated by decreases in BUN, UP, and renal index. In addition, consistent with the morphological changes of renal tissue, renal interstitial fibrosis in CKD rats after RC intervention was significantly improved, mainly manifested by a decrease in the positive expression of COL-1, FN, and a-SMA, and increased levels of E-cadherin protein. Meanwhile, RC reduced the classical pro-inflammatory cytokines IL-1β, IL-6, and TNF-α in adenine-induced CKD rats. Additionally, RC administration also down-regulated TGF-β1, JNK, p38 and ERK.In conclusion, RC may reduce inflammation in adenine induced CKD rats, improve extracellular matrix (ECM) components deposition, and diminish epithelial-mesenchymal transition (EMT) marker protein levels. Furthermore, RC intervention significantly reduces the release of inflammatory cytokines and inhibits the TGF-β1/MAPK signaling pathway. Based on the results, RC might be useful in the treatment of adenine induced renal fibrosis.Copyright © 2023. Published by Elsevier B.V.