Oxysophoridine

Oxymatrine inhibits melanoma development by modulating the immune microenvironment and targeting the MYC/PD-L1 pathway.[Pubmed:37788594]

Int Immunopharmacol. 2023 Nov;124(Pt B):111000.

Oxymatrine, also known as ammothamnine or Oxysophoridine, is a natural compound isolated from Sophora flavescens (in Chinese, Kushen), and many previous researchers have characterized its anti-inflammatory, anti-fibrotic and anti-tumor properties. However, the underlying anti-tumor immunological mechanism of oxymatrine remains elusive. In this study, we carried out experiments both in vitro and in vivo and investigated the anti-tumor effect of oxymatrine to inhibit the proliferation and migration of melanoma B16 cells, while promoting apoptosis. Oxymatrine upregulated CD4(+) T, CD8(+) T and NKT cells, downregulated Treg cells, promoted TNF-alpha secretion, and successfully modulated the immune microenvironment and ultimately suppressed melanoma development in subcutaneous tumor models established in mice. Evidence from network pharmacology and RNAseq suggested that possible targets of oxymatrine for melanoma treatment included PD-L1 and MYC. We observed oxymatrine inhibited PD-L1 and MYC expression in melanoma cells via qRT-PCR and western blotting analysis, and found MYC potentially regulated PD-L1 to mediate anti-tumor effects. These findings provide insight into the mechanism by which oxymatrine inhibits melanoma and enhances the anti-tumor immune effect. In summary, our study proposes a novel approach to suppress melanoma by targeting the MYC/PD-L1 pathway using oxymatrine, which may develop into a less toxic and more efficient anti-tumor agent for melanoma treatment.

Oxysophoridine protects against cerebral ischemia/reperfusion injury via inhibition of TLR4/p38MAPK‑mediated ferroptosis.[Pubmed:36601753]

Mol Med Rep. 2023 Feb;27(2):44.

Oxysophoridine (OSR) is an alkaloid extracted from Sophora alopecuroides L. and exerts beneficial effects in cerebral ischemia/reperfusion (I/R) injury. However, the molecular mechanism underlying the regulatory effects of OSR in cerebral I/R injury remains unclear. In the present study, a cerebral I/R injury rat model was established by occlusion of the right middle cerebral artery. Hematoxylin and eosin and triphenyltetrazolium chloride staining were performed to assess histopathological changes and the extent of cerebral injury to the brain. A Cell Counting Kit‑8 and TUNEL assay and western blotting were performed to assess cell viability and apoptosis. Ferroptosis and oxidative stress were evaluated based on ATP and Fe(2+) levels and DCFH‑DA staining. The protein expression levels of inflammatory factors were assessed using ELISA. The protein expression levels of members of the toll‑like receptor (TLR)4/p38MAPK signaling pathway were evaluated using immunofluorescence staining and western blotting. The results demonstrated that OSR decreased brain injury and neuronal apoptosis in the hippocampus in I/R‑induced rats. OSR inhibited reactive oxygen species (ROS) production, decreased levels of ATP, Fe(2+) and acyl‑CoA synthetase long‑chain family member 4 (ACSL4) and transferrin 1 protein and increased the protein expression levels of ferritin 1 and glutathione peroxidase 4. Furthermore, OSR blocked TLR4/p38MAPK signaling in brain tissue in the I/R‑induced rat. In vitro experiments demonstrated that TLR4 overexpression induced generation of ROS, ATP and Fe(2+), which promoted the expression of ferroptosis‑associated proteins in hippocampal HT22 neuronal cells. The ferroptosis inducer erastin decreased the effects of OSR on oxygen‑glucose deprivation/reoxygenation (OGD/R)‑induced cell viability, oxidative stress and inflammatory response. Together, the results demonstrated that OSR alleviated cerebral I/R injury via inhibition of TLR4/p38MAPK‑mediated ferroptosis.

Inhibitory Effect of Salvia miltiorrhiza Extract and Its Active Components on Cervical Intraepithelial Neoplastic Cells.[Pubmed:35268683]

Molecules. 2022 Feb 27;27(5):1582.

The effective treatment of cervical intraepithelial neoplasia (CIN) can prevent cervical cancer. Salvia miltiorrhiza is a medicinal and health-promoting plant. To identify a potential treatment for CIN, the effect of S. miltiorrhiza extract and its active components on immortalized cervical epithelial cells was studied in vitro. The H8 cell was used as a CIN model. We found that S. miltiorrhiza extract effectively inhibited H8 cells through the CCK8 method. An HPLC-MS analysis revealed that S. miltiorrhiza extract contained salvianolic acid H, salvianolic acid A, salvianolic acid B, monomethyl lithospermate, 9'''-methyl lithospermate B, and 9'''-methyl lithospermate B/isomer. Salvianolic acid A had the best inhibitory effect on H8 cells with an IC(50) value of 5.74 +/- 0.63 muM. We also found that the combination of salvianolic acid A and Oxysophoridine had a synergistic inhibitory effect on H8 cells at molar ratios of 4:1, 2:1, 1:1, 1:2, and 1:4, with salvianolic acid A/Oxysophoridine = 1:2 having the best synergistic effect. Using Hoechst33342, flow cytometry, and Western blotting analysis, we found that the combination of salvianolic acid A and Oxysophoridine can induce programmed apoptosis of H8 cells and block the cell cycle in the G2/M phase, which was correlated with decreased cyclinB1 and CDK1 protein levels. In conclusion, S. miltiorrhiza extract can inhibit the growth of H8 cells, and the combination of salvianolic acid A (its active component) and Oxysophoridine has a synergistic inhibitory effect on H8 cells and may be a potential treatment for cervical intraepithelial neoplasia.

Vasorelaxation effect of oxysophoridine on isolated thoracicc aorta rings of rats.[Pubmed:34975120]

Chin J Physiol. 2021 Nov-Dec;64(6):274-280.

Oxysophoridine (OSR) is a main active alkaloid extracted from Sophora alopecuroides, which is a traditional Chinese herbal medicine that has been used widely. In this study, we used thoracic aorta rings isolated from Sprague-Dawley rats to explore the vasodilative activity of OSR and its potential mechanisms. The isolated rat thoracic aorta rings were used to observe the effects of different concentrations of OSR (0.4-2.0 g.L(-1)) on the resting normal rings and the phenylephrine precontracted endothelium-intact or endothelium-denudedisolated thoracic aorta rings, respectively. The interactions among OSR and barium chloride (BaCl(2)), tetraethylamine, 4-aminopyridine, glibenclamide (Gli), L-nitroarginine methyl ester (L-NAME), and cyclooxygenase (COX) inhibitor indomethacin (INDO) were evaluated. The experimental results show that OSR had no effect on the tension of resting vascular rings, but the vasodilating effect could be confirmed in a concentration-dependent manner on both endothelium-intact and endothelium-denuded vascular rings. This vasodilation effect of OSR on thoracic aorta vascular rings could be inhibited significantly by potassium channel blockers glibenclamide (Gli, 10 mumol.L(-1)) and 4-aminopyridine (4-AP, 5 mmol.L(-1)). In addition, vasodilatory effects of OSR were not inhibited in the presence of potassium channel blockers barium chloride (BaCl(2), 1 mmol.L(-1)) and tetraethylamine (TEA, 10 mmol.L(-1)), nitric oxide synthase inhibitor (L-NAME, 0.1 mmol.L(-1)) and COX inhibitor (INDO, 10 mumol.L(-1)). In conclusion, the vasodilatory effects of OSR on thoracic aorta rings is associated with K(V) and K(ATP).

Targeting the TLR4/NF-kappaB pathway in beta-amyloid-stimulated microglial cells: A possible mechanism that oxysophoridine exerts anti-oxidative and anti-inflammatory effects in an in vitro model of Alzheimer's disease.[Pubmed:34329731]

Brain Res Bull. 2021 Oct;175:150-157.

beta-amyloid (Abeta) accumulation is a major neuropathological characteristic of Alzheimer's disease (AD) and serves as an inflammatory stimulus for microglial cells. Oxysophoridine has multiple pharmacological effects, including anti-inflammatory and anti-oxidative activities. In view of this, the current study aimed to investigate the effects of Oxysophoridine on Abeta-induced activation of microglial BV-2 cells. Cell Counting Kit-8 assay showed that Oxysophoridine concentration-dependently attenuated Abeta-induced viability reduction of BV-2 cells. Abeta stimulation reduced the activities of glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD) and elevated malondialdehyde (MDA) content in BV-2 cells, but these effects were attenuated by Oxysophoridine. Oxysophoridine abolished Abeta-induced increase of mRNA expression, secretion, and protein expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta) in BV-2 cells. Additionally, western blot suggested that Oxysophoridine inhibited Abeta-induced activation of the toll-like receptor 4 (TLR4) and nuclear factor-kappa B (NF-kappaB) pathways in BV-2 cells. Inhibition of the TLR4/NF-kappaB pathway by TAK-242 enhanced the effects of Oxysophoridine on Abeta-induced viability reduction, oxidative stress, and inflammation in BV-2 cells. Taken together, Oxysophoridine suppressed Abeta-induced oxidative stress and inflammation in BV-2 cells by inhibition of the TLR4/NF-kappaB pathway.

Gemcitabine, lycorine and oxysophoridine inhibit novel coronavirus (SARS-CoV-2) in cell culture.[Pubmed:32432977]

Emerg Microbes Infect. 2020 Dec;9(1):1170-1173.

The emerging SARS-CoV-2 infection associated with the outbreak of viral pneumonia in China is ongoing worldwide. There are no approved antiviral therapies to treat this viral disease. Here we examined the antiviral abilities of three broad-spectrum antiviral compounds gemcitabine, lycorine and Oxysophoridine against SARS-CoV-2 in cell culture. We found that all three tested compounds inhibited viral replication in Vero-E6 cells at noncytotoxic concentrations. The antiviral effect of gemcitabine was suppressed efficiently by the cytidine nucleosides. Additionally, combination of gemcitabine with Oxysophoridine had an additive antiviral effect against SARS-CoV-2. Our results demonstrate that broad-spectrum antiviral compounds may have a priority for the screening of antiviral compounds against newly emerging viruses to control viral infection.

Matrine-Type Alkaloids from the Roots of Sophora flavescens and Their Antiviral Activities against the Hepatitis B Virus.[Pubmed:30298740]

J Nat Prod. 2018 Oct 26;81(10):2259-2265.

Eight new matrine-type alkaloids, flavesines G-J (1-4), alopecurine B (5), 7,11-dehydro-oxymatrine (6), 10-oxy-5,6-dehydromatrine (7), and 10-Oxysophoridine (8), along with nine known analogues (9-17) were isolated from the roots of Sophora flavescens. Compounds 1-3 are the first natural matrine-type alkaloids with an open-loop ring D, while compound 4 represents an unprecedented dimerization pattern constructed from matrine and piperidine, and 5 is the first example of a matrine-type alkaloid with cleavage of the C-5-C-6 bond. The new structures were elucidated by means of spectroscopic data analysis (including NMR, MS, IR, and UV), and the absolute configurations were determined using single-crystal X-ray diffraction and ECD data. The isolated alkaloids were evaluated for their antiviral activity against hepatitis B virus, and compounds 1, 4, 5, 10, and 14 exhibited comparable antiviral potencies to matrine.

In vivo and in vitro induction of the apoptotic effects of oxysophoridine on colorectal cancer cells via the Bcl-2/Bax/caspase-3 signaling pathway.[Pubmed:29344242]

Oncol Lett. 2017 Dec;14(6):8000-8006.

Oxysophoridine (OSR) is a major active alkaloid extracted from Sophoraalopecuroides L. The aim of the present study was to investigate the induction of the apoptotic effects of OSR on colorectal cancer cells in vivo and in vitro. The results of the MTT and colony formation assays demonstrated that the proliferation of HCT116 cells was inhibited by OSR in vitro. The characteristics of cellular apoptosis in OSR-treated HCT116 cells were analyzed by Hoechst 33258 staining. It was also observed that the expression of caspase-3, B-cell lymphoma-2 (Bcl-2) associated X protein (Bax) and cytochrome c increased significantly upon OSR treatment. However, the expression of Bcl-2 and poly ADP-ribose polymerase-1 (PARP-1) was downregulated in OSR-treated cells compared with untreated cells. The in vivo experiments identified that OSR significantly inhibited the growth of the transplanted mouse CT26 tumor tissue, upregulated the expression of caspase-3, Bax and cytochrome c and downregulated the expression of Bcl-2 and PARP-1, as detected by reverse transcription-quantitative polymerase chain reaction and western blotting. It may be concluded that OSR significantly induced apoptotic effects on colorectal cancer cells in vivo and in vitro, and that its mechanism may be associated with the Bcl-2/Bax/caspase-3 signaling pathway.

Oxysophoridine rescues spinal cord injury via anti‑inflammatory, anti‑oxidative stress and anti‑apoptosis effects.[Pubmed:29207118]

Mol Med Rep. 2018 Feb;17(2):2523-2528.

Oxysophoridine (OSR) is an alkaloid extracted from Sophora alopecuroides L and has various pharmacological activities. The present study aimed to investigate the protective effects and underlying mechanisms of OSR on spinal cord injury (SCI), a clinically common serious trauma, in a rat model. The results of the present study demonstrated that the anti‑inflammatory effect of OSR improved Basso, Beatie and Bresnahan Locomotor Rating Scale scores and reduced spinal cord tissue water contents in an SCI rat model. Inflammatory activation was measured by ELISA, and Prostaglandin E2 (PGE2), intercellular adhesion molecule‑1 (ICAM‑1), cyclooxygenase‑2 (COX‑2), nuclear factor‑kappaB (NF‑kappaB) and B‑cell lymphoma 2 (Bcl‑2)/Bcl‑2‑associated X (Bax) protein expression levels using western blotting. The results revealed that treatment with OSR reduced tumor necrosis factor‑alpha, interleukin (IL)‑1beta, IL‑6, IL‑8 and malondialdehyde, and increased superoxide dismutase and glutathione peroxidase levels in the serum of an SCI rat model. OSR significantly reduced the protein expression of inflammation‑associated proteins PGE2, ICAM‑1, COX‑2, NF‑kappaB and Bcl‑2/Bax ratio in the spinal cord tissue of an SCI rat model. Furthermore, the results of the current study demonstrate that OSR ameliorates SCI via anti‑inflammatory, anti‑oxidative stress and anti‑apoptosis effects.

Anti-inflammatory and anti-apoptotic effects of oxysophoridine on lipopolysaccharide-induced acute lung injury in mice.[Pubmed:26885265]

Am J Transl Res. 2015 Dec 15;7(12):2672-82. eCollection 2015.

Oxysophoridine (OSR) is an alkaloid with multiple pharmacological activities. This study aimed to investigate the protective effects and underlying mechanisms of OSR on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Here, we found that OSR treatment markedly mitigated LPS-induced body weight loss and significant lung injury characterized by the deterioration of histopathology, histologic scores, wet-to-dry ratio, exduate volume, and protein leakage. OSR dramatically attenuated LPS-induced lung inflammation, as evidenced by the reduced levels of total cells, neutrophils, lymphocytes, and macrophages and pro-inflammatory cytokines (i.e., tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, and monocyte chemoattractant protein-1) in bronchoalveolar lavage fluid and of their mRNA expression in lung tissues. OSR also inhibited LPS-induced expression and activation of nuclear factor-kappaB p65 in pulmonary tissue. Additionally, OSR administration markedly prevented LPS-induced pulmonary cell apoptosis in mice, as reflected by the decrease in expression of procaspase-8, procaspase-3, cleaved caspase-8, and cleaved caspase-3, and Bcl-2-associated X/B-cell lymphoma 2 ratio. These results indicate that OSR is a potential therapeutic drug for treating LPS-induced ALI.

Anti-inflammation Effects of Oxysophoridine on Cerebral Ischemia-Reperfusion Injury in Mice.[Pubmed:26178478]

Inflammation. 2015 Dec;38(6):2259-68.

Oxysophoridine (OSR) is a bioactive alkaloid extracted from the Sophora alopecuroides Linn. Our aim is to explore the potential anti-inflammation mechanism of OSR in cerebral ischemic injury. Mice were intraperitoneally pretreated with OSR (62.5, 125, and 250 mg/kg) or nimodipine (Nim) (6 mg/kg) for 7 days followed by cerebral ischemia. The inflammatory-related cytokines in cerebral ischemic hemisphere tissue were determined by immunohistochemistry staining, Western blot and enzyme-like immunosorbent assay (ELISA). OSR-treated groups observably suppressed the nuclear factor kappa B (NF-kappaB), intercellular adhesion molecule-1 (ICAM-1), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). OSR-treated group (250 mg/kg) markedly reduced the inflammatory-related protein prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and interleukin-8 (IL-8). Meanwhile, it dramatically increased the interleukin-10 (IL-10). Our study revealed that OSR protected neurons from ischemia-induced injury in mice by downregulating the proinflammatory cytokines and blocking the NF-kappaB pathway.

Oxysophoridine attenuates the injury caused by acute myocardial infarction in rats through anti-oxidative, anti-inflammatory and anti-apoptotic pathways.[Pubmed:25338622]

Mol Med Rep. 2015 Jan;11(1):527-32.

Oxysophoridine (OSR), a natural alkaloid derived from the traditional Chinese medicinal plant sophora alopecuroides, can perform a variety of pharmacological actions. The aim of the present study was to assess the cardioprotective effect of OSR against acute myocardial infarction (AMI) in rats. OSR markedly reduced infarction size and levels of specific myocardial enzymes, including creatine kinase, the MB isoenzyme of creatine kinase, lactate dehydrogenase and cardiac troponin T. A reduced level of malondialdehyde was observed, and elevated catalase, Cu/Zn-superoxide dismutase (SOD), Mn-SOD, non-enzymatic scavenger glutathione and glutathione peroxidase activity were also identified in the OSR-treated rats. Additionally, OSR inhibited the activities of various inflammatory cytokines in a dose-dependent manner. These included nuclear factor-kappaB p65, tumor necrosis factor-alpha, and interleukin-1beta, -6 and -10. Furthermore, OSR treatment suppressed caspase-3 activity in a dose-dependent manner. These results demonstrate that OSR ameliorates cardiac damage in a rat model of AMI and that this cardioprotection may be linked with its anti-oxidative, anti-apoptotic and anti-inflammatory properties.

Effects of oxysophoridine on amino acids after cerebral ischemic injury in mice.[Pubmed:25221402]

Ann Indian Acad Neurol. 2014 Jul;17(3):313-6.

BACKGROUND: Our previous studies demonstrated that Oxysophoridine (OSR) had neuroprotective effects on mice through antioxidant and anti-apoptotic mechanisms. In this study, we investigated whether OSR could influence the release of amino acids in ischemic mice brains. MATERIALS AND METHODS: Male ICR mice were scheduled to undergo 2 h middle cerebral artery occlusion (MCAO) and 24 h reperfusion. Before MCAO, mice in corresponding groups were intraperitoneally injected with OSR (62.5, 125 and 250 mg/kg) for seven successive days. After reperfusion, neurological scores were estimated, infarct volume and the brain water content were assessed. The levels of glutamate (Glu), aspartate (Asp), gamma-aminobutyric acid (GABA) and Glycine (Gly) were measured by amino acid analyzer. RESULTS: OSR significantly decreased neurological scores, reduced infarct volume and the brain water content. After treatment with OSR of 250 mg/kg, the contents of Glu, Asp, GABA and Gly in mice brains could maintain at a normal level compared with MCAO group mice. The Glu/GABA ratio was significantly decreased in OSR group mice. CONCLUSION: These findings indicate that OSR has a protective effect on cerebral ischemic injury and helps to maintain the amino acids homeostasis after reperfusion for a long time.

Oxysophoridine protects against focal cerebral ischemic injury by inhibiting oxidative stress and apoptosis in mice.[Pubmed:24078262]

Neurochem Res. 2013 Nov;38(11):2408-17.

Our previous studies have demonstrated that Oxysophoridine (OSR) has protective effects on cerebral neurons damage in vitro induced by oxygen and glucose deprivation. In this study, we further investigated whether OSR could reduce ischemic cerebral injury in vivo and its possible mechanism. Male Institute of cancer research mice were intraperitoneally injected with OSR (62.5, 125 and 250 mg/kg) for seven successive days, then subjected to brain ischemia induced by the model of middle cerebral artery occlusion. After reperfusion, neurological scores and infarct volume were estimated. Morphological examination of tissues was performed. Apoptotic neurons were detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling staining. Oxidative stress levels were assessed by measurement of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels. The expression of various apoptotic markers as Caspase-3, Bax and Bcl-2 were investigated by immunohistochemistry and Western-blot analysis. OSR pretreatment groups significantly reduced infract volume and neurological deficit scores. OSR decreased the percentage of apoptotic neurons, relieved neuronal morphological damage. Moreover, OSR markedly decreased MDA content, and increased SOD, GSH-Px activities. Administration of OSR (250 mg/kg) significantly suppressed overexpression of Caspase-3 and Bax, and increased Bcl-2 expression. These findings indicate that OSR has a protective effect on focal cerebral ischemic injury through antioxidant and anti-apoptotic mechanisms.