在这项研究中，我们探讨了脂联素调节泌尿激肽II（UII）诱导的肿瘤坏死因子-α（TNF-α）和α平滑肌肌动蛋白（α-SMA）表达及随之而来的内部信号通路在外膜成纤维细胞（AFs）中的情况。停滞生长的AFs和大鼠血管外膜培养物经过1-24小时的UII和信号转导通路抑制剂孵育。然后，利用反转录-聚合酶链式反应（RT-PCR）和酶联免疫吸附测定（ELISA），收集细胞进行TNF-α受体（TNF-α-R）mRNA和TNF-α蛋白表达检测。脂联素和脂联素受体（AdipoR）的表达通过RT-PCR、实时荧光定量PCR（qPCR）、免疫组化分析和CCK-8细胞增殖实验进行测量。然后通过qPCR和免疫荧光染色量化TNF-α和α-SMA mRNA和蛋白表达水平。采用RNA干扰（RNAi）技术来探究adipoR基因的功能。为了研究信号通路，我们应用西方印迹（WB）检测腺苷酸5’-单磷酸（AMP）激活的蛋白激酶（AMPK）的磷酸化。在体内，生成了一种脂联素（APN）敲除（APN-KO）小鼠模型，模拟外膜炎症，通过应用qPCR和IF，测量TNF-α和α-SMA的表达，以期获得脂联素在血管重塑中的全面图谱。 无论是在细胞还是组织中，UII通过Rho/protein激酶C（PKC）信号转导途径剂量和时间依赖地促进TNF-α蛋白和TNF-α-R的分泌。我们检测到在AFs中明显表达adipoR1、T-细胞蛋白质黏附分子（T-cadherin）和钙调蛋白，而adipoR2仅有适度存在，未观察到脂联素的存在。球形脂联素（gAd）促进了AFs的生长，并通过adipoR1/T-cadherin/钙调蛋白/AMPK信号途径与UII协同诱导α-SMA和TNF-α的发生。在AFs中，gAd和UII协同诱导AMPK的磷酸化。在外膜炎症模型中，APN缺陷上调了α-SMA、UII受体（UT）和UII的表达，同时抑制了TNF-α的表达。 从我们的研究结果可以推断出，UII通过Rho和PKC信号转导途径在AFs和大鼠血管外膜中诱导了TNFα蛋白和TNF-α-R的分泌。因此，推论出脂联素是外膜病变进展的主要因素，并可以作为心血管疾病治疗的新的治疗靶标。

In this study, we explored how adiponectin mediated urotensin II (UII)‍-induced tumor necrosis factor-‍α (TNF-‍α) and α‍-smooth muscle actin (α‍-SMA) expression and ensuing intracellular signaling pathways in adventitial fibroblasts (AFs).Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UII and inhibitors of signal transduction pathways for 1‍‒‍24 h. The cells were then harvested for TNF-α receptor (TNF-‍α-R) messenger RNA (mRNA) and TNF-‍α protein expression determination by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Adiponectin and adiponectin receptor (adipoR) expression was measured by RT-PCR, quantitative real-time PCR (qPCR), immunohistochemical analysis, and cell counting kit-8 (CCK-8) cell proliferation experiments. We then quantified TNF-α and α-SMA mRNA and protein expression levels by qPCR and immunofluorescence (IF) staining. RNA interference (RNAi) was used to explore the function of the adipoR genes. To investigate the signaling pathway, we applied western blotting (WB) to examine phosphorylation of adenosine 5'-monophosphate (AMP)‍-activated protein kinase (AMPK). In vivo, an adiponectin (APN)‍-knockout (APN-KO) mouse model mimicking adventitial inflammation was generated to measure TNF-α and α‍-SMA expression by application of qPCR and IF, with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling.In both cells and tissues, UII promoted TNF-α protein and TNF-α-R secretion in a dose- and time-dependent manner via Rho/protein kinase C (PKC) pathway. We detected marked expression of adipoR1, T-cadherin, and calreticulin as well as a moderate presence of adipoR2 in AFs, while no adiponectin was observed. Globular adiponectin (gAd) fostered the growth of AFs, and acted in concert with UII to induce α-SMA and TNF-α through the adipoR1/T-cadherin/calreticulin/AMPK pathway. In AFs, gAd and UII synergistically induced AMPK phosphorylation. In the adventitial inflammation model, APN deficiency up-regulated the expression of α-SMA, UII receptor (UT), and UII while inhibiting TNF-‍α expression.From the results of our study, we can speculate that UII induces TNF‍-‍α protein and TNF-‍α‍-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways. Thus, it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.