EXPERIMENTAL ANIMALS
·
2022
·
IF 1.798
Ning HAN,Ying SU,Mi GUO,Li YU
Diabetic retinopathy (DR) is the prevalent microvascular complication of diabetes mellitus (DM), and it may lead to permanent blindness. The previous publication has indicated that both inflammatory response and oxidative stress are critical factors involved in DR progression, however, the accurate regulatory mechanism remains to be revealed. Src homology region 2 (SH2)-containing protein tyrosine phosphatase 2 (SHP2), a member of the protein tyrosine phosphatase family, was reported to play a role in diabetic nephropathy, whereas its function in DR was unknown and required further exploration. The level of phosphorylated, not the total, SHP2 increased in the retinas of rats with streptozotocin injection-induced DM. Further, the intravitreal injection of SHP2 shRNA lentivirus alleviated retinal pathological changes, and inhibited inflammatory response and oxidative stress, which were accompanied with Yes-associated protein 1 (YAP1) deactivation in DR rats. Additional co-immunoprecipitation results confirmed the interaction of SHP2 and YAP1. Collectively, our data preliminarily show that DR amelioration-induced by SHP2 inhibition in rats may attribute to the deactivation of YAP1 pathway.
研究领域:
眼科学
·
分子生物学
·
糖尿病研究
·
细胞信号传导
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Aging-US
·
2024
·
IF 3.9
Yaming Yang,Zheng Yan,Qi Xie,Yong Wang,Zhiying Liu,Min Lei
Background: The purpose of this study is to observe LP45 ( Lactobacillus plantarum 45) to investigate the mechanism by which LP45 attenuates oxidative stress-induced damage and regulates the osteoblast-osteoclast balance. Materials and Methods: The oxidative stress level and osteoblast- and osteoclast-related proteins were detected by immunofluorescence staining, Western blotting, ROS fluorescent probe and ELISA. Osteoblast cell proliferation capacity was determined by the CCK-8 assay. X-ray observation and HE staining were used to detect the effect of LP45 on osteoporosis. Results: The expression level of SHP2 and Src was significantly increased, and the expression levels of NOX4, P22, P47, IL-1β, NLRP3, IRF3, RANK, β-catenin and INF-β were inhibited in LP45 group and LPS + LP45 group as compared to those in LPS group. Compared with that in LPS group, the concentration of SOD was increased and the concentration of MDA was decreased in LPS + LP45 group. The protein expressions of OPG, RANKL, RUNX3, RANK and β-catenin in LP45 group and LPS + LP45 group increased. The protein expressions of NF-κB, CREB and AP-1 in LP45 group and LPS + LP45 group decreased significantly. The results were also confirmed by immunofluorescence staining and ROS fluorescent probe. X-ray observation and HE staining showed that LP45 could inhibit the progression of osteoporosis. Conclusion: LP45 can exert its antioxidant effect by inhibiting the production of oxidative stress to activate the SHP2 signaling pathway, thus promoting osteoblast differentiation and repressing osteoclast formation to maintain bone homeostasis and improve bone metabolism.
研究领域:
微生物学
·
骨骼生物学
·
药理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
·
2023
·
IF 6.7
Ruixiang Luo,Weitao Fu,Jingjing Shao,Lin Ma,Sujuan Shuai,Ying Xu,Zheng Jiang,Zenghui Ye,Lulu Zheng,Lei Zheng,Jie Yu,Yawen Zhang,Lina Yin,Linglan Tu,Xinting Lv,Jie Li,Guang Liang,Lingfeng Chen
Src homology 2 domain-containing phosphatase 2 (SHP2) is a cytoplasmic protein tyrosine phosphatase (PTP) that regulates signal transduction of receptor tyrosine kinases (RTKs). Abnormal SHP2 activity is associated with tumorigenesis and metastasis. Because SHP2 contains multiple allosteric sites , identifying inhibitors at specific allosteric binding sites remains challenging. Here, we used structure-based virtual screening to directly search for the SHP2 “tunnel site” allosteric inhibitor. A novel hit (70) was identified as the SHP2 allosteric inhibitor with an IC 50 of 10.2 μM against full-length SHP2. Derivatization of hit compound 70 using molecular modeling-guided structure-based modification allowed the discovery of an effective and selective SHP2 inhibitor, compound 129, with 122-fold improved potency compared to the hit. Further studies revealed that 129 effectively inhibited signaling in multiple RTK-driven cancers and RTK inhibitor-resistant cancer cells. Remarkably, 129 was orally bioavailable ( F = 55%) and significantly inhibited tumor growth in haematological malignancy . Taken together, compound 129 developed in this study may serve as a promising lead or candidate for cancers bearing RTK oncogenic drivers and SHP2-related diseases.
研究领域:
癌症生物学
·
药理学
·
分子生物学
·
药物发现
✓ 已验证
DOI
PubMed
高影响因子
Scientific Reports
·
2026
·
IF 4.9
Zhang Wei,Zhu Xinghao,Shao Mingliang,Han Qi,Shen Hao,Zhang Kun,Chen Rui,Qi Dongmei,Xu Lijun,Wu Zongyao
The progression of hepatocellular carcinoma (HCC) was highly dependent on pathological angiogenesis, with tumor-associated macrophages (TAMs) serving as key mediators regulating angiogenesis within the tumor microenvironment. The role of protein tyrosine phosphatase 2 (SHP2), a multi-signaling pathway regulator containing an SH2 domain, in TAM-mediated angiogenesis remained unclear. This study aimed to elucidate the molecular mechanism by which SHP2 regulates HCC angiogenesis through the ROS/Src pathway in TAMs under hypoxic conditions. We employed a comprehensive approach. THP-1 macrophage cells were transfected with SHP2 shRNA lentivirus and co-cultured with HCC cell line Huh-7 cells. The co-cultured cells were then subcutaneously transplanted into nude mice. Tumor size was assessed via HE staining in nude mouse tumor formation assays. We engineered luciferase-transfected Huh-7 cells to establish an orthotopic HCC implantation model and measured tumor area via in vivo fluorescence imaging. THP-1 cells were respectively cultured in media supplemented with antioxidants, pro-oxidants, and Src inhibitors, followed by transfection with SHP2 shRNA lentivirus and SHP2 mock lentivirus. These cells were then co-cultured with Huh-7 cells and Hep3B cells, respectively. The proportion of CD68-positive cells detected by immunofluorescence staining was used to ensure that the observed effects in the study were primarily derived from tumor-associated macrophages. Western blot analysis was employed to detect expression levels of SHP2, ROS/Src pathway-related proteins, tumor migration-related proteins, and angiogenesis-related proteins in the cells. Relative fluorescence intensity of intracellular ROS in THP-1 cells was visualized using the fluorescent probe DCFH-DA combined with fluorescence microscopy. Cell migration and invasion capabilities were assessed via wound healing assays and transwell assays. Immunofluorescence staining further validated SHP2 expression levels in THP-1 cells. Additionally, we examined the effects of SHP2 on HCC angiogenesis using tube formation assays, Western Blot analysis, HE staining, and immunohistochemical staining for CD31. The impact of SHP2 on HCC cell proliferation was investigated through colony formation assays and cell cycle analysis by flow cytometry. Following co-culture of SHP2 shRNA-infected THP-1 cells with Huh-7 cells, nude mouse tumor formation assays and in vivo fluorescence imaging revealed significantly reduced tumor volume and area. Following antioxidant supplementation in THP-1 cell culture medium, western blot analysis revealed markedly reduced expression levels of ROS/Src pathway-associated proteins, tumor migration proteins, and angiogenesis-related proteins. Fluorescent probe-based fluorescence microscopy demonstrated decreased relative fluorescence intensity of ROS within tumor cells. Furthermore, Western blot and immunofluorescence staining experiments jointly demonstrated that SHP2 in TAMs can regulate the ROS/Src pathway. Wound healing assays, transwell assays, and tube formation assays demonstrated that SHP2 in TAMs enhanced HCC cell migration, invasion, and angiogenesis capabilities. Furthermore, HE staining and immunohistochemical staining for CD31 indicated that inhibiting ROS/Src pathway suppressed HCC angiogenesis. Colony formation assay and flow cytometry analysis revealed that SHP2 in TAMs promotes HCC proliferation and cell cycle progression via the ROS/Src pathway. Under hypoxic conditions, tyrosine phosphatase SHP2 in TAMs regulates angiogenesis in HCC through the modulation of the ROS/Src pathway.
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
·
2025
·
IF 4.7
Simin Wang,Xiuhong Weng,Chang Liu,Bo Cheng
Salivary gland (SG) dysfunction is a hallmark manifestation of Sjögren's disease (SjD). While SHP2 has been implicated in several autoimmune diseases, its specific function in SjD remains to be elucidated. Here, we investigated the involvement of SHP2 in SjD-induced SG damage and elucidate the underlying molecular mechanisms. Our findings revealed that SHP2 is elevated in SG tissues of SjD patients and SjD-like animals, with predominant localization to salivary gland epithelial cells (SGECs) of acini and ducts. Notably, pharmacological inhibition of SHP2 with the allosteric inhibitor SHP099 significantly alleviates SG histopathological damage and reduces inflammatory cell infiltration in SjD-like animals. In vitro studies in the human SGEC line A253 revealed that SHP2 inhibition attenuated poly(I:C)-induced SGEC apoptosis and proinflammatory cytokine production. Mechanistically, SHP099 treatment disrupted the IL-17RA/ACT1/TRAF6 signaling axis, phenocopying the effects of SHP2 knockdown and direct IL-17R antagonism, which leads to downregulation of JAK/STAT3 and NF-κB pathways. Collectively, our findings highlight the significant involvement of SHP2 in the pathogenesis of SjD and suggest that targeting SHP2 or its downstream signaling pathways may represent a promising therapeutic approach for SjD treatment.
✓ 已验证
DOI
PubMed
高影响因子
IMMUNITY
·
2023
·
IF 32.4
Qiong Luo,Yijun Liu,Ke Shi,Xuecheng Shen,Yaqi Yang,Xuejiao Liang,Liangliang Lu,Wenxuan Qiao,Airu Chen,Dongmei Hong,Yang Sun,Qiang Xu
Anti-interleukin-17 (IL-17) therapy has been used in various autoimmune diseases. However, the efficacy is unexpectedly limited in several IL-17-associated diseases, and the mechanism of limited efficacy remains unclear. Here, we show that a molecular complex containing the adaptor molecule Act1 and tyrosine phosphatase SHP2 mediated autonomous IL-17R signaling that accelerated and sustained inflammation. SHP2, aberrantly augmented in various autoimmune diseases, was induced by IL-17A itself in astrocytes and keratinocytes, sustaining chemokine production even upon anti-IL-17 therapies. Mechanistically, SHP2 directly interacted with and dephosphorylated Act1, which replaced Act1-TRAF5 complexes and induced IL-17-independent activation of IL-17R signaling. Genetic or pharmacologic inactivation of SHP2, or blocking Act1-SHP2 interaction, paralyzed both IL-17-induced and IL-17-independent signaling and attenuated primary or relapsing experimental autoimmune encephalomyelitis. Therefore, Act1-SHP2 complexes mediate an alternative pathway for autonomous activation of IL-17R signaling, targeting which could be a therapeutic option for IL-17-related diseases in addition to current antibody therapies.
研究领域:
免疫学
·
分子生物学
·
自身免疫疾病
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）