目的 探讨大黄-丹参药对（RS）调控肌腱蛋白C（TNC）/表皮生长因子受体（EGFR）/信号转导和转录活化因子3（STAT3）通路延缓单侧输尿管梗阻（UUO）大鼠肾纤维化的机制。方法 50只雄性SD大鼠随机分为假手术组、模型组、尿毒清颗粒(阳性药，2.5 g·kg^-1)组和RS低、高剂量(生药3.0、6.0 g·kg^-1)组，每组10只。除假手术组外，其余各组大鼠采用UUO法制备肾间质纤维化模型。各给药组ig相应药物，假手术组、模型组ig等体积纯净水，连续14 d。代谢笼收集各组大鼠24 h尿液，腹主动脉取血分离血清，测定肌酐（Scr）和尿素氮（BUN）水平；酶联免疫吸附法（ELISA）检测尿液和血清中TNC的水平；HE和Masson染色观察各大鼠肾组织病理形态学变化；实时荧光定量法（qRT-PCR）检测肾组织TNC、EGFR、STAT3、TGF-β、纤维连接蛋白（FN）和α-平滑肌肌动蛋白（α-SMA）的m RNA表达；蛋白免疫印记法（Western blotting）检测肾组织TNC、EGFR、STAT3和磷酸化STAT3（p-STAT3）的蛋白表达；免疫组化染色（IHC）观察肾组织TNC、EGFR、p-STAT3的阳性表达。SD大鼠ig RS (生药6.0 g·kg^-1)或纯净水制备含药或空白血清；以人肾皮质近曲小管上皮（HK-2）细胞为研究对象，将细胞分为对照组、模型组(TGF-β 10 ng·mL^-1)、RS含药血清（10%）(RS)组、过表达阴性对照（NC-OE）组、TNC过表达（TNC-OE）组、TNC过表达+RS含药血清（10%）(TNC-OE+RS)组。按相应条件培养后，qRT-PCR检测各组细胞TNC、EGFR和STAT3 mRNA的表达，Western blotting检测TNC、EGFR、STAT3和p-STAT3的表达。结果 体内实验结果表明，与假手术组相比，模型组大鼠血清Scr、BUN、TNC水平和尿液TNC水平均明显升高（P<0.01）；肾组织病理损伤加重，肾纤维化评分显著升高（P<0.01）；肾组织TNC、EGFR、STAT3、TGF-β、FN和α-SMA m RNA水平和TNC、EGFR、p-STAT3蛋白表达水平及其阳性表达占比均明显上调（P<0.01）。与模型组相比，RS低、高剂量组和尿毒清颗粒组大鼠血清Scr、BUN、TNC水平和尿液TNC水平均显著降低（P<0.01）；肾组织损伤减轻，纤维化评分明显降低（P<0.01）；肾组织TNC、EGFR、STAT3、TGF-β、FN和α-SMA m RNA水平和TNC、EGFR、p-STAT3蛋白表达水平及其阳性表达占比均显著下调（P<0.05、0.01）。细胞实验结果表明，与对照组相比，模型组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平明显升高（P<0.01）；与模型组相比，RS组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平显著降低（P<0.05、0.01）。与NC-OE组相比，TNC-OE组细胞TNC、EGFR、STAT3 m RNA表达及TNC、EGFR、p-STAT3蛋白表达水平明显升高（P<0.01）；与TNC-OE组相比，TNC-OE+RS组细胞TNC、EGFR、STAT3 mRNA表达及TNC、EGFR、p-STAT3蛋白表达水平显著降低（P<0.01）。结论 RS能够从体内、外延缓肾纤维化的进展，其作用机制可能与调控TNC/EGFR/STAT3通路有关。

Objective To investigate the mechanism by which the Rheum palmatum-Salvia miltiorrhiza dug pairs（RS） regulates the tendon protein C（TNC）/epidermal growth factor receptor（EGFR）/signal transducer and activator of transcription 3（STAT3） pathway in delaying renal fibrosis in unilateral ureteral obstruction（UUO） rats. Methods Fifty male SD rats were randomly divided into the sham-operated group, the model group, the urine Niaoduqing Granule(positive drug, 2.5 g·kg^-1) group, and the RS low-dose and highdose(raw drug 3.0 and 6.0 g·kg^-1) groups, with 10 rats in each group. Except for the sham-operated group, the other groups of rats were prepared to establish a renal interstitial fibrosis model by UUO. Each administration group was given the corresponding drug, while the sham-operated group and the model group were given the same volume of pure water. The drugs were administered for 14 consecutive days. Metabolic cages were used to collect the 24 h urine of each groups of rats, and blood was taken from the abdominal aorta to separate the serum. The levels of creatinine（Scr） and urea nitrogen（BUN） were measured; the levels of TNC in urine and serum were detected by enzyme-linked immunosorbent assay（ELISA）; HE and Masson staining were used to observe the morphological changes of renal tissue; real-time fluorescence quantitative method（qRT-PCR） was used to detect the mRNA expression of TNC, EGFR, STAT3, TGF-β, fibronectin（FN）, and α-smooth muscle actin（α-SMA） in renal tissue; Western blotting was used to detect the protein expression of TNC, EGFR, STAT3, and phosphorylated STAT3（p-STAT3）; immunohistochemical staining（IHC） was used to observe the positive expression of TNC, EGFR, and p-STAT3 in renal tissue. SD rats were given RS(raw drug 6.0 g·kg^-1) or pure water to prepare the drug-containing or blank serum; human renal cortical proximal tubular epithelial（HK-2） cells were used as the research object. The cells were divided into the control group, the model group(TGF-β 10 ng·mL^-1), the 10% RS drug-containing serum（RS） group, the negative control group（NC-OE）, the TNC overexpression group（TNC-OE）, and the TNC overexpression + RS drug-containing serum（10%）（TNC-OE + RS） group. After corresponding conditions of cultivation, qRT-PCR was used to detect the expression of TNC, EGFR, and STAT3 mRNA in each group of cells, and Western blotting was used to detect the expression of TNC, EGFR, STAT3, and p-STAT3. Results The results of the in vivo experiments showed that compared with the sham operation group, the levels of serum Scr, BUN, TNC, and urine TNC in the model group were significantly increased（P ＜ 0.01）; The renal tissue pathological damage was aggravated, and the renal fibrosis score was significantly increased（P ＜ 0.01）; The mRNA levels of TNC, EGFR, STAT3, TGF-β, FN, and α-SMA in renal tissue and the protein expression levels and positive expression ratios of TNC, EGFR, p-STAT3 were significantly upregulated（P ＜ 0.01）. Compared with the model group, the serum Scr, BUN, TNC levels, and urine TNC levels in the RS low-dose and high-dose groups and the urine Niaoduqing Granule group were significantly decreased（P ＜ 0.01）; the renal tissue damage was alleviated, and the fibrosis score was significantly reduced（P ＜ 0.01）; The mRNA levels of TNC, EGFR, STAT3, TGF-β, FN, and α-SMA in renal tissue and the protein expression levels and positive expression ratios of TNC, EGFR, p-STAT3 were significantly downregulated（P ＜ 0.05, 0.01）. The results of the cell experiments showed that compared with the control group, the mRNA expressions of TNC, EGFR, and STAT3 in the model group and the protein expressions of TNC, EGFR, and p-STAT3 were significantly increased（P ＜ 0.01）; Compared with the model group, the mRNA expressions of TNC, EGFR, and STAT3 and the protein expressions of TNC, EGFR, and p-STAT3 in the RS group were significantly decreased（P ＜ 0.05, 0.01）. Compared with the NC-OE group, the mRNA expressions of TNC, EGFR, and STAT3 and the protein expressions of TNC, EGFR, and p-STAT3 in the TNC-OE group were significantly increased（P ＜ 0.01）; Compared with the TNC-OE group, the mRNA expressions of TNC, EGFR, and STAT3 and the protein expressions of TNC, EGFR, and p-STAT3 in the TNC-OE + RS group were significantly decreased（P ＜ 0.01）. Conclusion RS can delay the progression of renal fibrosis both in vivo and in vitro, and its mechanism of action may be related to the regulation of the TNC/EGFR/STAT3 pathway.

R285.5

河北省中医药管理局科研计划项目（T2025032）