Current Molecular Pharmacology - Volume 14, Issue 5, 2021

Background: Pain is often flammable, sharp and sometimes described as an electrical shock. It can be categorized in three different ways as nociceptive, neuropathic and inflammatory. Nociceptive pain always originates in specific situations such as in trauma. Neuropathic pain results in nerve damage. In inflammatory pain, inflammatory mediators are involved in the sensitization of nociceptors. It is important to control the pain as it affects the individual physically, mentally, and socially. Objective: Recognizing pain physiopathology and pain pathways, defining the relationship between receptor and transmitter is critical in developing new treatment strategies. In this review, current information on the definitions, classifications, and physiological and chemical mechanisms involved in pain are reviewed. Methods: Various search engines were used to gather related articles/information. Only peer-reviewed journals were considered. Additional, books/chapters of standard publishers were also included in the article. Results: With a better understanding of the physiological and chemical mechanisms that play a role in pain, significant improvements have been made in pain treatment. Various oral or intravenous drugs, local injection treatments, physical and occupational therapy, electrical stimulation, alternative medicine applications, psychological support, and surgical applications are routinely performed in the treatment, dependent upon the type, severity and cause of the pain. Conclusion: Improved understanding of pain physiopathology will serve as the basis for future improvements in the delivery of efficacious and reliable treatments, and is likely to rely on novel technological innovations.

Tissue cells of the eye are sensitive to oxidative stress, leading to oxidative damage and inflammation, which have crucial roles in the pathogenesis of most of the eye diseases. Moreover, factors such as age, angiogenesis, ischemia, genetic predisposition, diabetes, obesity, and smoking affect the onset and progression of ocular anomalies. Major eye diseases are either age-related such as age-related macular degeneration, cataracts, diabetic retinopathy, glaucoma, dry eye, or inflammatory eye diseases such as blepharitis, conjunctivitis, uveitis and other retinal pathologies, which can cause ocular damage and visual problems through oxidative stress, inflammation pathways. Although there are conventional chemical or surgical treatments, due to their mainly antiapoptotic, anti- inflammatory and antioxidant properties, phytotherapeutic preparations and their natural compounds can be applied to prevent or treat eye disease as follows: oral, topical or both. Thus, this study aims to comprehensively evaluate the pre-clinical and clinical studies of the phytotherapeutics and natural compounds used in the prevention and treatment of common serious and painful eye diseases. Moreover, for the first time, nature-derived preparations/supplements commonly used for eye problems also included in this review to present applications of the extracts/compounds in the pharmaceutical field.

Spinal cord injury (SCI) and associated pain and inflammation caused by trauma or infection are serious health care issues world-wide. The various inflammatory, redox-sensitive and apoptotic events are contributing factors, but altered neuronal function, axonal degeneration, activated microglia, endothelial cells, astrocytes, fibroblasts, pericytes, Schwann cells, and meningeal cells are major players in its pathogenesis. Further, monocytes and neutrophil infiltration get recruited and facilitate the release of chemokines, cytokines, and other mediators of inflammation. This event leads to the production of different amino acids, neuropeptides kinin, prostaglandins, prostacyclin, thromboxane, leukotrienes, bradykinin, histamine, matrix metal proteinases, and serotonin that stimulate nerve endings and manifest the inflammation and pain processes, etc. Arachidonic acid (AA), NF-kB, NLRP3 inflammasome, and nitric oxide pathways along with P2X7 receptor and ion channel transient receptor potential (TRP) vanilloid are some of the recently explored targets for modulation of pain and inflammation in SCI. Till now, NSAIDs, opioids, antidepressants, anticonvulsants, NMDA antagonists, α2-adrenergic agonists, and GABA-receptor agonists are used for the management of these pathological conditions. However, these drugs are associated with various side effects. Additionally, the number of available animal models for SCI has enhanced the understanding of the complex pathological mechanisms involved in the generation of chronic inflammatory pain in SCI. These findings enable us to identify and validate several potent natural analgesic-anti-inflammatory drug candidates with minimal side effects. However, these compounds have been studied in preclinical models and shown promising results, but no clinical studies have been performed. Therefore, a detailed exploration of these natural compounds is important for bringing them from bench to bedside.

Osteoarthritis and rheumatoid arthritis are the most common diseases of the musculoskeletal system, which greatly reduce the quality of life of people. In addition to non-steroidal anti- inflammatory drugs used in treatment, molecules isolated from natural sources are also considered as new options in the treatment of these inflammatory diseases. In this review, in vitro, in vivo and clinical studies on standardized rosehip (Rosa canina L.) fruits without seed used for joint health due to their anti-inflammatory, analgesic and antioxidant effects and active compounds isolated from these fruits are presented. It is reported that the anti-inflammatory action mechanism of standardized rosehip powder is due to its antioxidant activity, inhibiting NF-B signaling and pro-inflammatory enzymes, decreasing inflammatory cytokine and chemokine production, and lowering C reactive protein levels. The galactolipid (2S)-1,2-di-O-[(9Z,12Z,15Z)-octadeca-9,12,15- trienoyl]-3-O-ß-D-galactopyranosyl glycerol (GOPO), isolated from rosehip seeds and fruits, has been found to exhibit potent anti-inflammatory effects in vitro, clinically reducing the complaints of patients with osteoarthritis and improving their quality of life. Additionally, triterpene acid mixture (ursolic acid, oleanolic acid, and betulinic acid), also isolated from rosehip, has been reported to reduce the production of interleukin-6 and Tumor necrosis factor-α. Studies on the anti-inflammatory mechanism of action of rosehip and its active ingredients and their effects on osteoarthritis and rheumatoid arthritis have shown that more detailed clinical studies are required on standardized rosehip powders and preparations enriched in active compounds.

Background: Numerous therapeutic agents are in clinical practice for the treatment of inflammatory and painful conditions, but their applications have been challenged by various side /toxic effects. Therefore, new effective and safe therapies are most warrant, for which medicinal plant could be a significant alternative. Berberis baluchistanica has traditionally been used as analgesic and anti-inflammatory without any scientific background. Objective: The current study was designed to evaluate the analgesic and anti-inflammatory like effects of extract B. baluchistanica in animal models. Methods: For the study of antinociceptive effect, an extractof B. baluchistanica (100, 200 and 300 mg/kg i.p.), was tested in acetic acid-induced writhing and formalin tests. While for the anti-inflammatory action, carrageenan-induced paw edema, cotton pellet induced granuloma and xylene induced ear edema tests were used. Results: The results showed the significant dose-dependent antinociceptive effect of extract of B. baluchistanica in acetic acid-induced writhing and formalin-induced flinching behavior tests. However, the effect was strongly antagonized by the injection of naloxone, suggesting the expression via opioidergic receptors. Similarly, strong anti-inflammatory action was illustrated in carrageenan- induced paw edema, cotton pellet induced granuloma and xylene induced ear edema tests and thus provided evidence for the versatile phytochemical nature of its phytochemical. Conclusion: In short, it is concluded that the extract of B. baluchistanica exhibited significant antinociceptive and anti-inflammatory effects in animal models. Further detailed studies on the efficacy and safety as well as on the phytochemical investigation are required to ascertain its clinical uses.

Background: Oriental planetree (Platanus orientalis L.) leaf was recorded as a remedy against inflammatory problems and to stop the pain, i.e. toothache or knee pain in “The Canon of Medicines” by Avicenna and was also being documented in the Iranian Traditional Medicine. Although such a utilization has not been described in reliable sources, recently use of its leaves as herbal tea has become popular among laypeople. Previous studies have shown that only the nonpolar extract from the leaves may have such efficacy, while possible benefits when it is used as herbal tea have not been investigated. Objective: The present study aims to reveal the possible efficacy and safety profile of aqueous extract from P. orientalis leaf. Methods: Aqueous extract of the leaves was submitted to in vivo and in vitro tests to determine its anti-inflammatory, antinociceptive, antimutagenic activities and also reveal its safety profile. Results: The aqueous extract (400 mg/kg) procured weak and non-significant anti-inflammatory and antinociceptive activities. Meanwhile, the aqueous extract demonstrated antimutagenic activity in very high concentrations. On the other hand, results of safety evaluation showed that no concern had been observed from the viewpoint of public health. Conclusion: Despite the popularity of the herbal tea prepared from the leaves of Oriental planetree among the people suffering joint problems to relieve pain, this study has proven that such application would not help them to alleviate their complaints when used as herbal tea.

Hepatocellular carcinoma (HCC) is one of the prominent forms of cancer in developed countries. Globally, the incidence of HCC is well correlated with fatty liver disease and cirrhosis; the underlying chronic inflammation and lipotoxicity are thought to drive the process of HCC. Several biochemical cycles and molecular pathways are associated. The review aims to summarize the role of PI3K/Akt signaling and its downstream effectors in the development of HCC and its progression. Further, the emphasis has been given to the role of natural inhibitors of the PI3K/Akt pathway in HCC prevention, which are under various levels of drug discovery. The required literature were collected from PubMed/Medline databases, as well as Scopus or Web of science. It is evident that various signaling pathways activated by growth factors together with detoxification machinery and biochemical cycles converge to the PI3K/Akt signaling. The pathway plays a key role in the carcinogenesis, metastasis and drug resistance events of HCC cells and provides the growth and survival advantage. Natural products belonging to various classes such as terpenoids, flavonoids, saponins and stilbenoids are proven inhibitors of PI3K signaling and also found to inhibit HCC progression. PI3K/mTOR pathway inhibitors, especially the different phytochemicals, emerged as promising as anti-HCC agents. These molecules are shown to interfere with the PI3K signaling at various stages and therefore the PI3K targeted drugs may be a future for the chemotherapeutic arena.

Mycotoxins are defined as fungi that have negative effects on human health. The immune system of the organism is affected by mycotoxins first, and developmental problems, systemic diseases and death can be observed. Although this damage is important for all individuals, due to immaturity of the developmental areas, weak immune system and vulnerability against external factors, it can seriously affect children both in the pre- and in the postpartum period. It is aimed to examine the effects of mycotoxins on child development, which is also very important in the study, together with the studies conducted. In the research, a document analysis method was used to determine the effects of mycotoxins on children. The articles examining the effects of mycotoxins and mycotoxins on children were searched from Google Scholar between 2002 and 2020, accessible books were examined and the results of these studies were discussed as a whole. As a result of the encounter of the fetus with mycotoxins in the womb, the rate of low birth weight babies increases. At the same time, growth retardation, congenital anomalies and neural tube defects are encountered in the fetus. In infancy, mycotoxins cause developmental retardation, damage to the immune system, health problems, especially gastrointestinal problems, and cognitive development problems. As a result of encountering mycotoxins in childhood and adolescence, developmental delays, cognitive and neurological problems, and academic failures can be seen.

Cardiac fibrosis is a maladaptive condition secondary to cardiomyopathy caused by a wide spectrum of stimuli, including myocardial infarction (MI), pressure overload, hyperglycemia, aging, and other factors. Despite having been supposed to be a reparative mechanism, the development of cardiac fibrosis can result in undesirable outcomes like the disruption of excitation-contraction coupling and ventricular hypertrophy, leading finally to heart failure (HF). Statins are known as potent cardioprotective agents widely used to control dyslipidemia; these drugs have exhibited protective effects against manifestations of cardiac fibrosis and hypertrophy. Cumulative evidence has suggested that statins attenuate the severity of fibrotic and hypertrophic manifestations of cardiac damage by affecting a variety of mechanisms like differentiation of myofibroblasts and crosstalk between cells in cardiac tissue as well as altering the expression and function of different molecules involved in cardiac remodeling, including inflammatory cytokines and signaling molecules. It seems that statins can inhibit cardiac fibrosis and hypertrophy not only through their ability to inhibit hydroxymethylglutaryl-CoA reductase but also by their pleiotropic properties. This review aims to discuss the effects of statins on molecular pathways involved in the inhibition of fibrotic and hypertrophic remodeling in the heart, thereby potentially helping to recover proper cardiac size, plasticity, and functioning.

Most of the tripartite motif (TRIM) family proteins have E3 ubiquitin ligase activities and also have a variety of functions in cellular processes such as intracellular signal transduction, apoptosis, innate immunity, and carcinogenesis. TRIM65 is a member of the TRIM family. More pieces of evidence have shown a unique role and importance of TRIM65 protein in the occurrence and development of some diseases. In this review, the importance of TRIM65 in white matter lesions, innate immunity, and the effect of tumors were mainly reviewed.

Background: Medicinal plants and herbal preparations in the form of traditional medicines have been used in healthcare worldwide. The extracts of Ginkgo biloba L. seeds and leaves contain a complex mixture of numerous components, such as flavonol glycosides, terpene lactones, and a group of alkylphenols (anacardic or ginkgolic acids, cardanols and cardols) that have been a part of traditional Chinese medicine. These extracts are also sold as dietary supplements worldwide. G. biloba extract (EGb 761 and LI 1370) represent the standard form of G. biloba extract. Six different 6-alkylsalicylic acids (syn. ginkgolic acids) with alkyl substituents (C13:0, C15:0, C15:1, C17:0, C17:1, and C17:2) have been identified. Objective: The aim of this review is to unravel scientific evidence on anti-inflammatory and anticancer activities of ginkgolic acids to understand its therapeutic potential against inflammatory and oncologic diseases. Methods: A structured literature search was independently performed by the authors on PubMed, ScienceDirect, Scopus, and Web of Science. Accordingly, this review article critically analyses available scientific evidence on anti-inflammatory and anticancer activities of ginkgolic acids. Moreover, the review only included articles written in the English language. Results: Several forms of ginkgolic acids, especially C13:0, C15:0 and C17:1, isolated from the leaves of G. biloba exhibited cytotoxic activity against a variety of human cancers by suppressing various pro-inflammatory signaling cascades and oncogenic transcription factors through multiple modes of action in various in vitro and in vivo preclinical models. Ginkgolic acids have also been reported to be potent post-translational small ubiquitin-related modifiers (SUMO)ylation inhibitors. Conclusion: In this review, we present updated information on the anti-inflammatory and anticancer properties of ginkgolic acids both in vitro and in vivo. Although ginkgolic acids show significant therapeutic potential in inflammatory and oncologic diseases, more investigations regarding the safety and efficacy of these natural agents are warranted before the clinical transition.

Multiple Sclerosis (MS) is a severe brain and spinal cord condition with a diverse autoimmune response and a wide variety of demyelination symptoms that primarily affect young adults. The primary reason for this disease is inflammation of white and grey matter caused by increased production of proinflammatory cytokines, which further damages the progenitor oligodendrocytes and appears to induce hypertrophy of the astrocytes and gliosis. Overexpression of the JAK/STAT signaling pathway contributes directly to physiological and pathological results in motor neuron diseases. Cytokines such as IL-17, IL-6, IL-12, TNF-α, and INF- use JAK/STAT signaling to trigger self-reactive CD4+ T-cells and differentiate them into Th1 phenotypes that overactivate immune reactions in the brain. Similarly, PPARγ plays a critical role in regulating the immune response by providing an anti-inflammatory effect by inhibiting macrophage and cytokine production activation. PPARγ also mediates the intrinsic molecular process of the T-cell, which selectively regulates the differentiation of Th17. Various studies indicate the neuroprotective function of PPARγ agonists by attenuating the JAK/STAT mediated activation of glial cells, inhibiting interleukin, and the differentiation of Th1 cells. Therefore, to maintain the brain's immune system, both PPARγ and JAK/STAT oppositely regulate each other. Dysregulation in JAK/STAT and PPARγ signaling contributes to several physiological changes leading to neurological disorders, including MS. Based on the above view, we have summarized the combined role of JAK/STAT-PPARγ signaling in MS and explored potential therapeutic strategies for disease improvement by the use of pathway modulators.

The recent developments in epigenetics have shown a very important role of epigenetic changes in cancer initiation, development, and progression. Some of the important histone modifications shown to occur are methylation, acetylation, phosphorylation, citrullination, sumoylation, ADP ribosylation, deamination, ubiquitination, formylation, O-GlcNAcylation, propionylation, butyrylation, proline isomerization, and crotonylation, but most of the studies in the past had limited their interest mainly on histone methylation, acetylation, and phosphorylation. Modification of DNA strands by hypermethylation and hypomethylation regulates genomic instability and promotes cancer. Colorectal cancer involves multiple changes in epigenetic marks present on histone residues and DNA, which in collaboration with genetic changes, leads to cancer progression. In this review paper, basic concepts of epigenetics relevant to cancer development are discussed, followed by its significance in understanding the mechanism of colon carcinogenesis. Some of the epigenetic target-based drugs are also discussed in the relevant sections to give an idea of the potential promises of epigenetics for colorectal cancer treatment.

Background and Objective: Exposure to mycotoxins may delay and/or negatively influence the development of neurological, gastrointestinal and inflammatory mechanisms in individuals with Autism Spectrum Disorder (ASD). Therefore, there is a need to address the possible links between mycotoxins and the risk and prevalence of ASD to increase the understanding of the molecular mechanism underlying these links. In this context, the aim of this study was to investigate the molecular mechanism underpinning mycotoxin exposure and autism. Methods: The study was based on a systematic approach, which focused on the possible associations between mycotoxins and ASD in addition to the role of the mycotoxins on the risk and prevalence of ASD. The systematic review included all molecular mechanism studies examining mycotoxin exposure and autism, and was not limited to a specific period of time. A search was performed on the PubMed, Web of Science, Scopus, and Google Scholar databases. Results: The investigation of the literature revealed that a total number of 11 studies with a specific focus on the molecular mechanism of mycotoxin exposure and autism were published between 2008 and 2019. Out of these studies, 7 were research articles and 4 were review articles. In almost all the articles, possible links between mycotoxins and ASD were revealed. Conclusion: The examination of the given studies provided data related to the links between mycotoxins and ASD. However, evidence related to these links needs to be investigated in larger samples, while the effects of separate mycotoxins and their metabolisms should also be examined.

Background: Androgens potentially have an important role in the biology of breast cancer, particularly triple-negative breast cancer (TNBC). Androgen receptor (AR) may offer a novel therapeutic strategy, including the use of microRNA (miRNA) molecules. We have previously shown that AR agonist, dihydrotestosterone (DHT), increases the expression of miR-328-3p in the TNBC MDA-MB-231 cells. One target of the latter miRNA is ATP-binding cassette subfamily G member 2 (ABCG2), which modulates the chemo-response of cancer cells by pumping out xenobiotics. Objective: Using MDA-MB-231 cells as a model system for TNBC, we hypothesized that DHT would induce cell sensitivity towards doxorubicin via increasing levels of miR-328-3p and, consequently, reducing ABCG2 levels. Methods: Chemo-response of cells towards doxorubicin, tamoxifen, and mitoxantrone was evaluated using cell viability MTT assay. Cells were transfected with both miR-328-3p mimic or antisense molecules. Real-time PCR was utilized to assess RNA levels and immunoblotting was performed to investigate levels of ABCG2 protein. PCR arrays were used to assess changes in the expression of drug response regulatory genes. Results: Contrary to our hypothesis, treating MDA-MB-231 cells with DHT no effect towards tamoxifen or mitoxantrone, increased cell resistance towards doxorubicin was noted, concomitant with decreased expression of ABCG2. This under-expression of ABCG2 was also found in MCF-7 and MDA-MB-453 cells treated with DHT. Although miR-328-3p decreased ABCG2 mRNA and protein levels, the miRNA did not alter the chemo-response of cells towards doxorubicin and did not affect DHT-induced chemo-resistance. AR activation slightly decreased the expression of 5 genes, including insulin-like growth factor 1 receptor that may explain the mechanism of DHT-induced chemo-resistance of cells. Conclusion: DHT regulates chemo-response via a mechanism independent of ABCG2 and miR-328-3p.

Background: Alcoholic fatty liver disease (AFLD), a leading chronic hepatic disease, affects an increasing number of people, and no effective drugs for the treatment of AFLD are available. Antrodia cinnamomea (AC) can inhibit AFLD, but its mechanisms and the effective compound in AC are unknown. Objective: We aimed to explore the anti-AFLD mechanism of AC and the active compound within AC. Methods: Wild-type (WT) C57BL/6J mice underwent 4 weeks of daily ethanol (EtOH) feeding to induce AFLD. AC or dehydroeburicoic acid 32 (DEA32), a compound in AC, was given to the mice. Parallel experiments to assess the effect of AC were conducted in aldehyde dehydrogenase 2 (ALDH2)-knockout (KO) mice. Primary mouse hepatocytes were incubated with ethanol and Alda- 1 (an ALDH2 agonist), AC or DEA32. Results: In WT mice with AFLD, AC reduced lipid deposition, increased the expression and activity of ALDH2, reduced the acetaldehyde content, and downregulated the expression of lipogenic and inflammatory genes in the liver. These effects of AC disappeared in ALDH2 KO mice. DEA32 was identified as an active compound in AC, as its effects on EtOH-treated WT hepatocytes were similar to those of AC, which were comparable to the effects of Alda-1. These effects of DEA32 disappeared in EtOH-treated ALDH2 KO hepatocytes. Furthermore, in WT mice with AFLD, DEA32 reduced lipid deposition, increased the activity of ALDH2, and reduced the accumulation of acetaldehyde in the liver. DEA32 also downregulated the mRNA expression of genes related to lipogenesis and inflammation. Conclusion: AC and its constituent compound DEA32 inhibit AFLD by upregulating ALDH2 activity, accelerating acetaldehyde metabolism, and suppressing lipogenesis and inflammation in the liver.

Background: Diabetic retinopathy (DR) is one of the most common side effects of diabetes. We aimed to investigate the effects of crocin and crocetin (as a deglycosylated form of crocin in blood stream) in gene expression or protein levels of vascular endothelial growth factor (VEGF), vascular endothelial growth factor-receptor1 (VEGFR-1), matrix metalloproteinases2 (MMP-2), matrix metalloproteinases9 (MMP-9) and thrombospondin-2 (TSP-2) in high glucose cell culture media. Methods: The retinal pigment epithelium (RPE) cells were exposed to high glucose (HG, 30 mM glucose concentration) and normal glucose (NG, 24.5 mM mannitol + 5.5 mM glucose) for six days. RPE cells were treated in four treatment groups (crocin, crocetin, Bevacizumab, and crocin + Bevacizumab). Gene expressions were measured using quantitative real-time PCR, and protein levels were evaluated by western blot. Results: Findings showed that VEGF gene expression and protein level significantly decreased in all treatment groups. In addition, reduction in VEGFR1 gene expression was significantly higher in Bevacizumab and crocin + Bevacizumab groups than other groups. Only crocin and crocetin could reduce the gene levels of MMP-2 and MMP-9. In addition, TSP-2 protein levels increased when HG cells were exposed to crocin or crocin + Bevacizumab groups. Conclusion: Our data showed that crocin and crocetin have anti-VEGF function similar to Bevacizumab, act as an anti-angiogenic agent. Also, crocin and crocetin could decrease MMP-2 and MMP-9 gene levels being inflammatory and angiogenesis factors. As a result, crocin and crocetin have protective effects against angiogenesis and inflammation in DR.

Background: Age-dependent toxic effects of organophosphorus pesticides (OPs) have not been fully understood. The current study aimed to investigate the cardiotoxic damage of chlorpyrifos (CPF) by evaluating oxidative modifications in young (2-month old), middle-aged (10- month old), and aged (20-month old) rats. Objective: Five mg/kg of CPF was administered orally for 45 days to young, middle-aged, and aged male Wistar rats. In the end, animals were anesthetized and the heart of each rat was dissected for biochemical assay. Methods: Malondialdehyde (MDA), nitric oxide (NO), glutathione (GSH), total antioxidant capacity (TAC), and superoxide dismutase (SOD) were assessed in the cardiac tissue of rats. Results: The results indicated an increase in the levels of MDA and NO, and also a decline in the levels of GSH and TAC as well as a decrease in the SOD activity in the heart of aged rats compared with young rats. CPF administration deteriorated these changes in the heart of exposed rats compared with the age-matched controls. Additionally, these oxidative modifications were more severe in aged rats versus other age. Conclusion: In conclusion, advancing age may increase oxidative changes in the heart of animals exposed to CPF. It is suggested that aging can affect cardiac toxicity induced by OPs.

Background: Hepatocellular carcinoma (HCC) is the 6^th prevalent cancer and the 4^th leading cause of cancer-related deaths all over the world. A major challenge for sorafenib, the standard chemotherapeutic agent in HCC treatment, is the chemo-resistance. Objective: This study was conducted to evaluate the role of dantrolene as a possible antineoplastic agent in HCC, and in chemo-sensitization of sorafenib via targeting Ca⁺²/PI3K pathway. Methods: HCC was induced in rats using a single dose of diethylnitrosamine (DENA) (200 mg/kg, ip), followed by phenobarbital sodium (0.05%) in drinking water for 18 weeks. At the end of the 18^th week, rats were allocated into 4 groups (10 rats/each), one group was left without treatment (DENA group) and the other three groups were treated with either sorafenib, dantrolene, or their combination for further 4 weeks. One day after the last injection, serum and liver tissues were collected. Liver tissue p53, VEGF, MMP-9, Cyclin D1, PI3K, and, serum AFP were assessed using immunoassay. Hepatic and serum Ca⁺² were also computed. Furthermore, Ki-67 was assessed immunohistochemically. Results: Dantrolene exhibited synergistic effect when used in combination with sorafenib compared to either drug alone (p <0.05) through decreasing p53, VEGF, MMP-9, Cyclin D1, and Ki-67. In addition, dantrolene was evidenced to have an inhibitory effect on Ca⁺²/PI3K pathway that mediates its antineoplastic action when used alone or in combination with sorafenib. Conclusion: Dantrolene exerted antineoplastic effect as well as augmented sorafenib antineoplastic activity via the intervention of Ca⁺²/PI3K pathway, manifested by ameliorating angiogenesis, apoptosis, proliferation and metastasis.