Targeting ATP2B1 impairs PI3K/Akt/FOXO signaling and reduces SARS-COV-2 infection and replication.

ATP2B1 is a known regulator of calcium (Ca2+) cellular export and homeostasis. Diminished levels of intracellular Ca2+ content have been suggested to impair SARS-CoV-2 replication. Here, we demonstrate that a nontoxic caloxin-derivative compound (PI-7) reduces intracellular Ca2+ levels and impairs SARS-CoV-2 infection. Furthermore, a rare homozygous intronic variant of ATP2B1 is shown to be associated with the severity of COVID-19. The mechanism of action during SARS-CoV-2 infection involves the PI3K/Akt signaling pathway activation, inactivation of FOXO3 transcription factor function, and subsequent transcriptional inhibition of the membrane and reticulum Ca2+ pumps ATP2B1 and ATP2A1, respectively. The pharmacological action of compound PI-7 on sustaining both ATP2B1 and ATP2A1 expression reduces the intracellular cytoplasmic Ca2+ pool and thus negatively influences SARS-CoV-2 replication and propagation. As compound PI-7 lacks toxicity in vitro, its prophylactic use as a therapeutic agent against COVID-19 is envisioned here.

Long-chain polyphosphates impair SARS-CoV-2 infection and replication.

Inorganic polyphosphates (polyPs) are linear polymers composed of repeated phosphate (PO4 3-) units linked together by multiple high-energy phosphoanhydride bonds. In addition to being a source of energy, polyPs have cytoprotective and antiviral activities. Here, we investigated the antiviral activities of long-chain polyPs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In molecular docking analyses, polyPs interacted with several conserved amino acid residues in angiotensin-converting enzyme 2 (ACE2), the host receptor that facilitates virus entry, and in viral RNA-dependent RNA polymerase (RdRp). ELISA and limited proteolysis assays using nano- LC-MS/MS mapped polyP120 binding to ACE2, and site-directed mutagenesis confirmed interactions between ACE2 and SARS-CoV-2 RdRp and identified the specific amino acid residues involved. PolyP120 enhanced the proteasomal degradation of both ACE2 and RdRp, thus impairing replication of the British B.1.1.7 SARS-CoV-2 variant. We thus tested polyPs for functional interactions with the virus in SARS-CoV-2-infected Vero E6 and Caco2 cells and in primary human nasal epithelial cells. Delivery of a nebulized form of polyP120 reduced the amounts of viral positive-sense genomic and subgenomic RNAs, of RNA transcripts encoding proinflammatory cytokines, and of viral structural proteins, thereby presenting SARS-CoV-2 infection in cells in vitro.

Enhanced inhibitory effect of ultra-fine granules of red ginseng on LPS-induced cytokine expression in the monocyte-derived macrophage THP-1 cells.

Red ginseng is one of the most popular traditional medicines in Korea because its soluble hot-water extract is known to be very effective on enhancing immunity as well as inhibiting inflammation. Recently, we developed a new technique, called the HAC-gearshift system, which can pulverize red ginseng into the ultra-fine granules ranging from 0.2 to 7.0 microm in size. In this study, the soluble hot-water extract of those ultra-fine granules of red ginseng (URG) was investigated and compared to that of the normal-sized granules of red ginseng (RG). The high pressure liquid chromatographic analyses of the soluble hot-water extracts of both URG and RG revealed that URG had about 2-fold higher amounts of the ginsenosides, the biologically active components in red ginseng, than RG did. Using quantitative RT-PCR, cytokine profiling against the Escherichia coli lipopolysaccharide (LPS) in the monocyte-derived macrophage THP-1 cells demonstrated that the URG-treated cells showed a significant reduction in cytokine expression than the RG-treated ones. Transcription expression of the LPS-induced cytokines such as TNF-alpha, IL-1beta, IL-6, IL-8, IL-10, and TGF-beta was significantly inhibited by URG compared to RG. These results suggest that some biologically active and soluble components in red ginseng can be more effectively extracted from URG than RG by standard hot-water extraction.

Inhibition of nitric oxide and tumor necrosis factor-alpha by moutan cortex in activated mouse peritoneal macrophages.

Moutan cortex (MC) is one of the most widely used Oriental herbal medicines for treating inflammatory diseases. In this study, the effect of MC on lipopolysaccharide (LPS) and recombinant interferon-gamma (rIFN-gamma)-induced production of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha were examined using mouse peritoneal macrophages. MC inhibited the LPS/rIFN-gamma-induced expression of inducible nitric oxide synthase (iNOS) and TNF-alpha release. To clarify the mechanism involved, the effect of MC on the activation of nuclear factor (NF)-kappaB was examined. The LPS/rIFN-gamma-induced activation of NF-kappaB was almost completely blocked by MC at 0.5 mg/ml. These findings demonstrate that the inhibition of the LPS/rIFN-gamma-induced production of NO and TNF-alpha by MC is due to the inhibition of NF-kappaB activation.

The effects of BR003 on memory and cell proliferation in the dentate gyrus of rat hippocampus.

BR003 is a multi-herbal formula that contains twelve medicinal herbs. We investigated the effects of oral administration of BR003 to Wistar rats on (a) learning and memory using a passive avoidance test and (b) cell proliferation in the dentate gyrus (DG) of the hippocampus using immunohistochemical analysis of 5-bromo-2-deoxyuridine (BrdU) expression. In the passive avoidance test, the retention time of the BR003-treated group was significantly longer than that of the control group (182.64+/-39.88 vs. 73.08+/-29.30 s, respectively; n=11; p<0.05). There were significantly more BrdU-immunoreactive cells in the DG in the BR003-treated group than in the control group (1281.07+/-151.16 vs. 818.01+/-132.98 cells per DG, respectively; n=11; p<0.05). These results suggest that the administration of BR003 not only improves learning and memory but also increases cell proliferation in the DG of the rat hippocampus.

Inhibition of lipopolysaccharide and interferon-gamma-induced expression of inducible nitric oxide synthase and tumor necrosis factor-alpha by Lithospermi radix in mouse peritoneal macrophages.

Lithospermi radix (LR, root of Lithospermum erythrorhizon Siebold. et Zuccarinii) has been used to treat various conditions, such as septic shock, eczema and burns. In this study, the effect of LR on lipopolysaccharide (LPS) and recombinant interferon-gamma (rIFN-gamma)-induced production of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha were examined using mouse peritoneal macrophages. At 0.01-1 mg/ml, LR inhibited the LPS/rIFN-gamma-induced expression of inducible nitric oxide synthase (iNOS) and TNF-alpha release. To clarify the mechanism involved, the effect of LR on the activation of nuclear factor (NF)-kappaB was examined. The LPS/rIFN-gamma-induced activation of NF-kappaB was almost completely blocked by LR at 1mg/ml without cytotoxicity. These findings demonstrate that the inhibition of the LPS/rIFN-gamma-induced production of NO and TNF-alpha by LR involves the inhibition of NF-kappaB activation.

Inhibition of production of reactive oxygen species and gene expression profile by treatment of ethanol extract of Moutan Cortex Radicis in oxidative stressed PC12 cells.

Moutan Cortex Radicis (MCR) is one of the most widely used Oriental medicines. In this study, we assessed the reducing effect of ethanol extract of MCR on hydrogen peroxide-induced reactive oxygen production, the main cause of cell damage or death in PC12 cells. The viability of cells treated with 1 mg/ml of MCR was significantly restored from that of oxidative-stressed PC12 cells. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H(2)DCFDA assay. MCR at 1-0.01 mg/ml concentration inhibited ROS production in oxidative-stressed cells. To identify candidate genes responsible for the anti-oxidative effects of MCR on PC12 cells, an oligonucleotide microarray analysis was performed. The result of gene expression profiles showed that 10 genes were up-regulated and 7 were down-regulated in MCR plus hydrogen peroxide treated cells compared with hydrogen peroxide treated cells. Among them, heme oxygenase (HO) and cathechol-O-methyltransferase (COMT) are related to regulation of ROS generation and the others are known to regulate cell survival and progression. Subsequently, we performed real-time RT-PCR to quantify the ROS related gene. MCR treatment increased the expression of HO by 370% and COMT by 280% at the concentration of 1 mg/ml. These findings suggest that MCR inhibits the production of ROS and cytotoxicity by oxidative-stressed PC12 cells through over-expression of HO and COMT.

Yukmijihwang-tang derivatives enhance cognitive processing in normal young adults: a double-blinded, placebo-controlled trial.

Yukmijihwang-tang (YMJ), also known as Luweidihuang-tang in China, has been widely used as a general herbal tonic for hundreds of years in many Asian countries. This study examines whether YMJ derivatives (YMJd) enhance cognitive ability in normal human subjects and discusses its potential as treatment for dementia patients with deficient cognitive ability. Subjects were divided into two groups, the placebo-treated group (n = 15) and the YMJd-treated group (n = 20). K-WAIS tests, a Korean version of an individual intelligence quotient (IQ) test, and a P300 latency assessment of event-related potential (ERP) were conducted in order to measure changes in cognitive ability before and after 6 weeks of YMJd treatment. The K-WAIS mean scores of the group treated with YMJd were significantly higher than those of the placebo group (p < 0.05), and their mean P300 latency was substantially shorter (p < 0.005). These results suggest that YMJd treatment accelerates the speed of information processing and enhances cognitive ability. YMJd treatment may help dementia patients or the elderly recover from cognition deficiencies or degeneration in clinic.

Taraxacum officinale protects against cholecystokinin-induced acute pancreatitis in rats.

AIM: Taraxacum officinale (TO) has been frequently used as a remedy for inflammatory diseases. The aim of this study was to investigate the effect of TO on cholecystokinin (CCK)-octapeptide-induced acute pancreatitis in rats. METHODS: TO at 10 mg/kg was orally administered, followed by 75 microg/kg CCK octapeptide injected subcutaneously three times after 1, 3 and 5 h. This whole procedure was repeated for 5 d. We determined the pancreatic weight/body weight ratio, the levels of pancreatic HSP60 and HSP72, and the secretion of pro-inflammatory cytokines. Repeated CCK octapeptide treatment resulted in typical laboratory and morphological changes of experimentally-induced pancreatitis. RESULTS: TO significantly decreased the pancreatic weight/body weight ratio in CCK octapeptide-induced acute pancreatitis. TO also increased the pancreatic levels of HSP60 and HSP72. Additionally, the secretion of IL-6 and TNF-alpha decreased in the animals treated with TO. CONCLUSION: TO may have a protective effect against CCK octapeptide-induced acute pancreatitis.

Anti-obesity effect of PM-F2-OB, an anti-obesity herbal formulation, on rats fed a high-fat diet.

PM-F2-OB is one of the most well-known traditional herbal medicines that are frequently used for the treatment of obesity in Korea. The anti-obesity effect of PM-F2-OB on rats fed a high-fat diet was investigated through analyses of changes in body weight, kidney fat weight, and blood biochemicals including cholesterol, free fatty acid, BUN, creatinine, HDL, LDL, phospholipids, SGOT, SGPT, total lipids, and triglycerides. The subjects in this study were divided into four groups: a normal group with a standard diet (N); a PM-F2-OB treatment group fed a standard diet (N+PM-F2-OB); a control group fed a high-fat diet (C); and a PM-F2-OB treatment group fed a high-fat diet (C+PM-F2-OB). There were no significant differences in body weight change between the N and N+PM-F2-OB treatments. Also, there was no significant difference in the amount of food intake between the C and C+PM-F2-OB treatments. These results suggest that PM-F2-OB has no significant toxicity and does not induce a dislike for that diet due to its smell or taste. Rats were administered a high-fat diet (20% (w/w)) for six weeks to induce obesity. The study shows that PM-F2-OB significantly prevented increases in body weight, cholesterol, LDL and total lipids that resulted from the high-fat diet. PM-F2-OB also decreased kidney fat weight and free fatty acid, phospholipid, and triglyceride concentrations induced by the high-fat diet to level equals or below the normal diet group. It was concluded from the results that PM-F2-OB has a distinct anti-obesity effect.