Goand Umesh K, Patel Inklisan, Verma Saurabh, Yadav Shubhi, Maity Debalina, Singh Naveen, Vishwakarma Sachin, Rathaur Shivam, Garg Richa, Gayen Jiaur R
Pharmaceutics & Pharmacokinetics Division, CSIR-Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.
Pharmaceutics & Pharmacokinetics Division, CSIR-Central Drug Research Institute, Lucknow 226031, India.
Cytokine. 2023 Nov;171:156354. doi: 10.1016/j.cyto.2023.156354. Epub 2023 Sep 5.
Pancreastatin, a dysglycemic hormone that encourages inflammation and steatosis in a variety of metabolic disorder animal models. The purpose of this study is to determine the effect of the pancreastatin inhibitor PSTi8 on immunometabolic changes in the liver of MCD-induced NASH mice.
Methionine and choline-deficient (MCD) diet was used for the development of NASH. Liver enzymes like SGOT, SGPT, and ALP and lipid profiles were also performed in the serum. Further, immunophenotyping study was performed in the liver through flowcytometer. Subsequently, Hematoxylin and Eosin, Picro Sirius Red and Masson's Trichrome staining were done to check the liver morphology and collagen staining, respectively. Inflammatory cytokines were measured through ELISA and gene expression through RT-PCR. The expression of α-SMA was examined using immunohistochemistry and immunofluorescence staining.
PSTi8 inhibited the expression of lipogenic genes in the liver and attenuated bad cholesterol, SGOT, SGPT, and ALP in the serum. PSTi8 improved the liver morphology and attenuated collagen deposition. Subsequently, PSTi8 attenuated inflammatory M1-macrophages, CD8T, CD4T cells and increased anti-inflammatory M2 macrophages, T-reg and eosinophil populations in the liver. It also attenuated the expression of pro-inflammatory genes like Mcp1, Tnfα, and Il6. Apart from this, PSTi8 attenuated the oxidative stress marker, like ROS, and MDA and fibrosis marker α-SMA in the liver. It also decreased the apoptosis and ROS and MDA level in the liver.
Overall, these compressive studies revealed that PSTi8 exhibited beneficial effect on the liver of MCD-induced NASH mice by attenuating inflammation and oxidative stress.
胰抑制素是一种血糖异常激素，在多种代谢紊乱动物模型中会促进炎症和脂肪变性。本研究旨在确定胰抑制素抑制剂PSTi8对蛋氨酸和胆碱缺乏（MCD）诱导的非酒精性脂肪性肝炎（NASH）小鼠肝脏免疫代谢变化的影响。
采用蛋氨酸和胆碱缺乏（MCD）饮食诱导NASH。还检测了血清中的肝酶，如谷草转氨酶（SGOT）、谷丙转氨酶（SGPT）和碱性磷酸酶（ALP）以及血脂水平。此外，通过流式细胞仪对肝脏进行免疫表型分析。随后，分别进行苏木精和伊红染色、苦味酸天狼星红染色和Masson三色染色，以检查肝脏形态和胶原染色情况。通过酶联免疫吸附测定（ELISA）检测炎性细胞因子，通过逆转录聚合酶链反应（RT-PCR）检测基因表达。使用免疫组织化学和免疫荧光染色检测α-平滑肌肌动蛋白（α-SMA）的表达。
PSTi8抑制肝脏中脂肪生成基因的表达，并降低血清中坏胆固醇、SGOT、SGPT和ALP的水平。PSTi8改善了肝脏形态并减少了胶原沉积。随后，PSTi8减少了肝脏中炎性M1巨噬细胞、CD8T细胞、CD4T细胞，并增加了抗炎性M2巨噬细胞、调节性T细胞和嗜酸性粒细胞的数量。它还减少了促炎基因如单核细胞趋化蛋白1（Mcp1）、肿瘤坏死因子α（Tnfα）和白细胞介素6（Il6）的表达。除此之外，PSTi8降低了肝脏中的氧化应激标志物，如活性氧（ROS）和丙二醛（MDA）以及纤维化标志物α-SMA。它还降低了肝脏中的细胞凋亡以及ROS和MDA水平。
总体而言，这些综合研究表明，PSTi8通过减轻炎症和氧化应激，对MCD诱导的NASH小鼠的肝脏具有有益作用。
