Xie Zhuo Tiao Zhi formula modulates intestinal microbiota and liver purine metabolism to suppress hepatic steatosis and pyroptosis in NAFLD therapy.

BACKGROUND: Current evidence indicates a rising global prevalence of Non-Alcoholic Fatty Liver Disease (NAFLD), which is closely associated to conditions such as obesity, dyslipidemia, insulin resistance, and metabolic syndrome. The relationship between the gut microbiome and metabolites in NAFLD is gaining attention understanding the pathogenesis and progression of dysregulated lipid metabolism and inflammation. The Xie Zhuo Tiao Zhi (XZTZ) decoction has been employed in clinical practice for alleviating hyperlipidemia and symptoms related to metabolic disorders. However, the pharmacological mechanisms underlying the effects of XZTZ remain to be elucidated. PURPOSE: The objective of this study was to examine the pharmacological mechanisms underlying the hypolipidemic and anti-inflammatory effects of XZTZ decoction in a mouse model of NAFLD, as well as the effects of supplementing exogenous metabolites on PO induced cell damage and lipid accumulation in cultured hepatocytes. METHODS: A high-fat diet (HFD) mouse model was established to examine the effects of XZTZ through oral gavage. The general condition of mice and the protective effect of XZTZ on liver injury were evaluated using histological and biochemical methods. Hematoxylin and eosin staining (H&E) staining and oil red O staining were performed to assess inflammatory and lipid accumulation detection, and cytokine levels were quantitatively analyzed. Additionally, the study included full-length 16S rRNA sequencing, liver transcriptome analysis, and non-targeted metabolomics analysis to investigate the relationship among intestinal microbiome, liver metabolic function, and XZTZ decoction. RESULTS: XZTZ had a significant impact on the microbial community structure in NAFLD mice. Notably, the abundance of Ileibacterium valens, which was significantly enriched by XZTZ, exhibited a negative correlation with liver injury biomarkers such as, alanine transaminase (ALT) and aspartate transaminase (AST) activity. Moreover, treatment with XZTZ led to a significant enrichment of the purine metabolism pathway in liver tissue metabolites, with inosine, a purine metabolite, showing a significant positive correlation with the abundance of I. valens. XZTZ and inosine also significantly enhanced fatty acid ß-oxidation, which led to a reduction in the expression of pro-inflammatory cytokines and the inhibition of liver pyroptosis. These effects contributed to the mitigation of liver injury and hepatocyte damage, both in vivo and vitro. Furthermore, the utilization of HPLC fingerprints and UPLC-Q-TOF-MS elucidated the principal constituents within the XZTZ decoction, including naringin, neohesperidin, atractylenolide III, 23-o-Acetylalisol B, pachymic acid, and ursolic acid which are likely responsible for its therapeutic efficacy. Further investigations are imperative to fully uncover and validate the pharmacodynamic mechanisms underlying these observations. CONCLUSION: The administration of XZTZ decoction demonstrates a protective effect on the livers of NAFLD mice by inhibiting lipid accumulation and reducing hepatocyte inflammatory damage. This protective effect is mediated by the upregulation of I.valens abundance in the intestine, highlighting the importance of the gut-liver axis. Furthermore, the presesnce of inosine, adenosine, and their derivatives are important in promoting the protective effects of XZTZ. Furthermore, the in vitro approaching, we provide hitherto undocumented evidence indicating that the inosine significantly improves lipid accumulation, inflammatory damage, and pyroptosis in AML12 cells incubated with free fatty acids.

Inosine in Neurodegenerative Diseases: From the Bench to the Bedside.

Neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS), currently represent major unmet medical needs. Therefore, novel therapeutic strategies are needed in order to improve patients' quality of life and prognosis. Since oxidative stress can be strongly involved in neurodegenerative diseases, the potential use of inosine, known for its antioxidant properties, in this context deserves particular attention. The protective action of inosine treatment could be mediated by its metabolite urate. Here, we review the current preclinical and clinical studies investigating the use of inosine in AD, PD, ALS, and MS. The most important properties of inosine seem to be its antioxidant action and its ability to raise urate levels and to increase energetic resources by improving ATP availability. Inosine appears to be generally safe and well tolerated; however, the possible formation of kidney stones should be monitored, and data on its effectiveness should be further explored since, so far, they have been controversial. Overall, inosine could be a promising potential strategy in the management of neurodegenerative diseases, and additional studies are needed in order to further investigate its safety and efficacy and its use as a complementary therapy along with other approved drugs.

Elevated Urate Levels Do Not Alter Bone Turnover Markers: Randomized Controlled Trial of Inosine Supplementation in Postmenopausal Women.

OBJECTIVE: Observational studies have consistently demonstrated that serum urate level positively correlates with bone mineral density (BMD). We undertook this study to determine whether moderate hyperuricemia induced by inosine supplements influences bone turnover markers in postmenopausal women over a 6-month period. METHODS: One hundred twenty postmenopausal women were recruited for a 6-month randomized, double-blind, placebo-controlled trial. Key exclusion criteria were osteoporosis, previous fragility fracture, bisphosphonate therapy, gout, kidney stones, and a urine pH level of ≤5.0. Participants were randomized in a 1:1 ratio to receive placebo or inosine. The coprimary end points were change in levels of N-propeptide of type I procollagen (PINP) and change in levels of ß-C-telopeptide of type I collagen (ß-CTX). Change in BMD, as measured by dual x-ray absorptiometry, was an exploratory end point. RESULTS: Administration of inosine led to a significant increase in serum urate concentration over the study period (P < 0.0001 for all follow-up time points). At week 26, the mean change in serum urate concentration was +0.13 mmoles/liter (+2.2 mg/dl) in the inosine group and 0.00 mmoles/liter (0 mg/dl) in the placebo group. There was no difference in PINP or ß-CTX levels between groups over the 6 months. There were no significant changes in bone density between groups over the 6 months. Adverse events and serious adverse events were similar between the 2 groups. CONCLUSION: This clinical trial shows that although inosine supplementation leads to sustained increases in serum urate levels over a 6-month period, it does not alter markers of bone turnover in postmenopausal women. These findings do not support the concept that urate has direct biologic effects on bone turnover.

[Radiation protectors within the radiation safety system for extended duration exploration missions].

Radiation environment in extended duration exploration missions is scrutinized in the context of the probability of the risks of deterministic and stochastic effects of radiation. Though the probability of severe radiation damage due to solar flare is very low, nonetheless it is requisite that the crew must be provided with appropriate, including pharmacological safeguards. The current nomenclature of radiation protectors composes short-term agents against acute radiation damage. Among the others, preparation B-190 is distinguished by particularly high effectiveness and universal action, and good tolerance even when organism is exposed to the extreme factors of space flight Regimen of B-290 therapy alone and with combination with aminothiol preparations have been developed to render treatment following multiple solar events. Effectiveness of radioprotectors can be increased substantially by local shielding of the abdomen and pelvis. The most promising nonspecific stimulators of total resistance of organism are riboxin (inosin) and combined preparation aminotetravit as well as vitamins tocopherol and retinol. Therapy combining B-190 with riboxin and aminotetravit is also under discussion. Cytokine neipogen is also viewed as a candidate agent for early therapy. Concern is raised about possible development of chronic oxidative stress in long-duration exploration missions. Highlighted is the significance of adequate nutrition supplemented with fresh vegetables as a source of the most valuable bioflavonoids. Antioxidants L-selenomethionine and melatonin proved their effectiveness against heavy nuclei of galactic radiation. An open issue is how to make natural antioxidants beneficial to oxidative stress control and attenuation of low-intensity galactic radiation.

The treatment of multiple sclerosis with inosine.

OBJECTIVE: The objective of this study is to evaluate the safety and tolerability of inosine in patients with relapsing-remitting multiple sclerosis (RRMS). The secondary objectives are to assess the effects of inosine administration on serum urate (UA) levels, the progression of neurologic disability, the cumulative number of new, active lesions on magnetic resonance imaging (MRI), and changes in serum levels for markers of inflammation. DESIGN: Oral administration of inosine was used to raise serum levels of the natural peroxynitrite scavenger UA in 16 patients with RRMS during a 1-year randomized, double-blind trial. OUTCOME MEASURES: The endpoints studied were relapse rate, disability assessed by the Kurtzke Expanded Disability Status Scale (EDSS), MRI, and analysis of serum levels of nitrotyrosine, and oxidative and pro-inflammatory makers. RESULTS: Increased serum UA levels correlated with a significant decrease in the number of gadolinium-enhanced lesions and improved EDSS. A number of MRI intensity-based parameters were altered by inosine treatment, in certain cases correlating with changes in serum UA levels. In a patient with low serum UA and high lesion activity, raising UA levels by inosine treatment decreased serum nitrotyrosine while increasing the ratio of Th2 to Th1 cytokines in circulating cells. The only side-effect correlated with inosine treatment was kidney stone formation in 4/16 subjects. CONCLUSIONS: These data suggest that the use of inosine to raise serum UA levels may have benefits for at least some MS patients. The effect of this treatment is likely to be a consequence of inactivation of peroxynitrite-dependent free radicals. Close monitoring of serum UA levels as well as other measures are required to avoid the potential development of kidney stones.

Simultaneous application of two neurotrophic factors after spinal cord injury.

We have studied the application of voltage gradients to injured spinal cord which enhanced regeneration of axons and reduced their retrograde degeneration after injury. This led to an implanted electronic device producing electrical fields sufficient to induce regeneration in both ascending and descending tracts of white matter (called oscillating field stimulation [OFS]), which has been associated with behavioral recovery in animal models of spinal cord injury (SCI). OFS has also proven to benefit neurologically complete spinal cord injured dogs and humans in clinical trials. These studies, however, have failed to confirm benefit if applied after the sub-acute period of SCI. Here we report on combining OFS with the application of a non-toxic neurotrophic factor, inosine, using a behavioral model for "chronic" SCI, the cutaneous trunci muscle (CTM) reflex in adult guinea pigs. Inosine was delivered subcutaneously in guinea pigs for 28 days using implantable "osmotic pumps"--alone or in combination with OFS. In all animals, experimental and control treatments were withheld for three months after a right lateral hemisection of the thoracic spinal cord. Both inosine and the combination therapy produced a statistically significant recovery of CTM receptive fields silenced permanently by spinal cord hemisection in controls--though the combination therapy enhanced the time of the appearance of recovered regions of skin. Retransection of the cord in three recovered animals eliminated the CTM recovery confirming changes in neural connections were restricted to the cord and not due to changes in cutaneous peripheral innervation. Morphometry of anterogradely labeled white matter revealed a statistically enhanced regeneration of ascending and descending projections in animals treated with the combination "therapy" compared to inosine alone. These data suggest that combining neurotrophic factors of differing modes of action likely enhance the outcome from "chronic" SCI.

Clinical study on Ganbi decoction in treating antituberculotic agent-caused liver injury.

OBJECTIVE: To study the effect and mechanism of Ganbi decoction (GBD) in treating patients with antituberculotic agent caused liver injury (ATB-LI). METHODS: One hundred and twenty-eight patients with ATB-LI were randomly assigned to the treated group (n = 66) and the control group (n = 62) with the envelop method. Meanwhile, 60 healthy persons were selected as the healthy control group. The treated group was treated by GBD one dose every day with the constituents modified depending on patients' symptoms, and the control group was treated with glucuronolactone tablets and inosine injection. One week was taken as one treatment course. The changes of clinical syndromes, physical signs, T-lymphycyte sub-groups and serum level of nitric oxide (NO) were observed before and after treatment and the recovery time of liver function was recorded. The outcome was compared with that in the healthy control group. RESULTS: In the treated group, 28 patients (42.4%) were cured, 30 (45.5%) improved and 8 (12.1%) ineffectively cured, the total effective rate being 87.9% (58/66). In the control group, 17 patients (27.4%) were cured, 24 (38.7%) improved, and 21 (33.9%) ineffectively cured, the total effective rate being 66.1% (41/62). The total effective rate in the treated group was significantly higher than that in the control group (P < 0.05). Liver function was improved in both groups, recovery time in the treated group was 12.0 +/- 7.0 days, which was significantly shorter than that in the control group (16.0 +/- 8.0 days), showing significant difference between the two groups (P < 0.05). The levels of CD3, CD4 and CD8 were significantly higher and level of NO significantly lower in the two groups of patients than those in the healthy control group (P < 0.05), but these parameters were improved more significantly in the treated group after treatment, when compared with those before treatment or with those in the control group, all showing significant difference (P < 0.05). CONCLUSION: GBD could prevent ATB-LI, and its mechanism could be by way of reducing NO production induced by endotoxin of macrophage and stimulating the proliferation of T-lymphycyte to elevate immunity.

[Clinical observation on prevention of "jia wei si jun zi tang" from damage of hepatic reserving function after intervention of liver cancer].

OBJECTIVE: To observe prevention of "Jia Wei Si Jun Zi Tang" (JWSJZT) from damage of hepatic reserving function after intervention of liver cancer. METHODS: 65 patients with advance stage of liver cancer were randomly into two groups, the control group (regular protective therapy) and the treatment group (regular protective therapy + JWSJZT). There were 33 and 32 cases in the control group and treatment group,respectively. ICGR15 was detected before the first and the second intervention treatment and one month after the second intervention treatment, respectively. ICGR15 change was observed before and after the intervention treatment in the two groups. RESULTS: ICGR15 was 11.13% +/- 7.28% in the treatment group and 11.81% +/- 7.43% in the control group (P > 0.05) before the first intervention; 18.36% +/- 9.82% in the control group and 12.18% +/- 5.22% in the treatment group, before the second invention treatment (P < 0.05); 19.12% +/- 9.96% in the control group and 12.83% +/- 5.28% in the treatment group one month after the second intervention treatment (P < 0.05). CONCLUSION: The western medicine + JWSJZT group is superior in curative effect to western medicine group.

Therapeutic intervention in experimental allergic encephalomyelitis by administration of uric acid precursors.

Uric acid (UA) is a purine metabolite that selectively inhibits peroxynitrite-mediated reactions implicated in the pathogenesis of multiple sclerosis (MS) and other neurodegenerative diseases. Serum UA levels are inversely associated with the incidence of MS in humans because MS patients have low serum UA levels and individuals with hyperuricemia (gout) rarely develop the disease. Moreover, the administration of UA is therapeutic in experimental allergic encephalomyelitis (EAE), an animal model of MS. Thus, raising serum UA levels in MS patients, by oral administration of a UA precursor such as inosine, may have therapeutic value. We have assessed the effects of inosine, as well as inosinic acid, on parameters relevant to the chemical reactivity of peroxynitrite and the pathogenesis of EAE. Both had no effect on chemical reactions associated with peroxynitrite, such as tyrosine nitration, or on the activation of inflammatory cells in vitro. Moreover, when mice treated with the urate oxidase inhibitor potassium oxonate were fed inosine or inosinic acid, serum UA levels were elevated markedly for a period of hours, whereas only a minor, transient increase in serum inosine was detected. Administration of inosinic acid suppressed the appearance of clinical signs of EAE and promoted recovery from ongoing disease. The therapeutic effect on animals with active EAE was associated with increased UA, but not inosine, levels in CNS tissue. We, therefore, conclude that the mode of action of inosine and inosinic acid in EAE is via their metabolism to UA.

[Use of antihypoxants in the acute period of myocardial infarction]. / Primenenie antigipoksantov v ostrom periode infarkta miokarda.

A total of 620 patients with acute myocardial infarction were followed up in order to assess the efficacy of antihypoxants as a component of intensive care. 385 of these patients, divided into groups of 20-40 subjects, were administered one of 12 antihypoxants or sessions of hyperbaric oxygenation during the acute period of the disease, the rest were treated traditionally. Analysis of clinical, laboratory, and prognostic values showed the highest protective effect of amtizol, lithium hydroxybutyrate, piracetam, and ubiquinone. Cytochrome C, riboxine, mildronate, and olifen were somewhat less active, and solcoseryl, bemitil, trimethasidine, and aspisol were the least effective. The protective potentialities of standard sessions of hyperbaric oxygenation were virtually null. The author proposes a parameter D, reflecting the difference between actual and predicted mortality, and the rating (score) system for assessing the routine laboratory diagnostic tests to be used together with the known criteria for evaluation of the protective effects of antihypoxants in patients with acute myocardial infarction.

Efeitos do alopurinol sobre as alteraçöes morfológicas produzidas pela isquemia testicular temporária em ratos / Reperfusion injury with allopurinol after short term testicular ischemia in rats

O objetivo do presente estudo foi avaliar, em ratos, se o alopurinol é capaz de proteger o testículo contra os efeitos da isquemia com duraçäo de uma hora, seguida de reperfusäo. Dezoito ratos adultos, Wistar-EPM1, com peso variando entre 300 e 340g, foram divididos em três grupos: controle (G1), isquemia (G2) e isquemia com alopurinol (G3) com seis animais cada. Após anestesia com pentobarbital sódico intraperitoneal (30mg/Kg), o testículo esquerdo era exteriorizado e mantido imerso em soluçäo salina (0,9 por cento) à temperatura ambiente durante uma hora. Nos grupos G2 e G3 o pedículo era clampeado durante o experimento (1h). No grupo G3 cada animal recebeu 50 mg/Kg de alopurinol V.O. nos dois dias anteriores e duas horas antes da isquemia. Após 60 dias os animais foram anestesiados, sacrificados e os testículos removidos, pesados e preparados para exame histopatológico (HE). As lâminas foram analisadas de acordo com a classificaçäo de Cosentino. Observou-se diminuiçäo significante do peso do testículo esquerdo nos grupos G2 e G3 (p<0,05) e o escore histopatológico mostrou grau importante de sofrimento testicular esquerdo nos grupos G2 e G3 (p<0,05). Concluiu-se que o alopurinol näo foi capaz de proteger o testículo do rato contra os efeitos do fenômeno de isquemia/reperfusäo

[Clinical and experimental study of 236 ED nutritional supportive therapy and inosine nutrition protection in postoperative gastric cancer patients].

To confirm the effect of nutritional supportive therapy on cancer patients, 52 postoperative gastric cancer patients were selected and given 236 ED as nutritional therapy. Body weight and nutritional index before and after operation, complication, duration in hospital were compared in the experimental group and control. The results showed that there was a significant difference between the two groups. When inosine was given with 236 ED to 30 mice bearing S 180 sarcoma, tumor weight in the treated mice was less than that of the control; level of amino-acids which can stimulate tumor growth decreased whereas c-AMP content in the tumor tissue increased. Based on the above results, a rational nutritional support protocol for tumor patients and a pathomorphological classification of tumor in high, moderate and low nutritional status are proposed.

[Stimulation of reparative processes in myocardial infarction: comparative evaluation of the effects of piracetam and inosine]. / Stimuliatsiia reparativnykh protsessov pri infarkte miokarda: sravnitel'naia otsenka vliianiia piratsetama i inozina.

A study of 102 patients with primary macrofocal myocardial infarction, admitted to hospital within the first 6 hours after the onset of the disease, and experimental evidence obtained in 110 Wistar rats are reported. In a control group of myocardial infarction patients, collagen synthesis was depressed in the presence of the activation of free-radical lipid oxidation and circulatory hypoxia. Piracetam and inosine treatment was associated with the inhibition of free-radical lipid oxidation and a reduction of hypoxia, those being accompanied by increased collagenogenesis and accelerated postinfarction scar formation. Piracetam, as compared to inosine, had a more marked stimulating effect on the development of compensatory myocardial hypertrophy, thus contributing to rapid recovery of cardiac contractility and reducing manifestations of circulatory insufficiency.

[Comparison of the antihypoxic effect of alpha-tocopherol, NAD and inosine during prolonged artificial circulation]. / Sopostavlenie protivogipoksicheskogo éffekta al'fa-tokoferola, NAD i inozina pri dlitel'nom iskusstvennom krovoobrashchenii.

Mathematical analysis was used to compare the antihypoxic action of alpha-tocopherol, inosine and oxidized NAD under long-term extracorporeal circulation. Of the drugs studied, oxidized NAD form was demonstrated to be the most active, inosine appeared less active, and alpha-tocopherol still less active. It is advisable to study more thoroughly the effect of oxidized NAD under long-term extracorporeal circulation.

Synthesis and antitumor activity of 7- and 9-(6'-deoxy-alpha-L-talofuranosyl) hypoxanthine and 9-(6'-deoxy-alpha-L-talofuranosyl)-6-thiopurine.

Reaction of 6-deoxy-2,3,5-tris-O-(p-nitrobenzoyl)-L-talofuranosyl bromide (1) with the trimethylsilyl derivative of hypoxanthine, followed by removal of blocking groups, afforded 9- (6) and 7-(6'-deoxy-alpha-L-talofuranosyl)hypoxanthine (7). A study of the published optical rotations and circular dichroic (CD) spectra of pentofuranosylpurines and of (6'-deoxy-beta-D-allo- and -alpha-L-talofuranosyl)purines prepared here suggests that the sign of rotation and the sign of the longer wavelength Cotton effect is determined solely by the configuration of C-1' and its position of attachment to the purine ring. For C-1' R nucleosides, the sign is negative for N-9-linked purine nucleosides and positive for the N-7-linked isomers, and vice versa for C-1'S purine nucleosides. Reaction of 1 with the trimethylsilyl derivative of 6-chloropurine afforded 4, which upon treatment with thiourea and deblocking yielded 9-(6'-deoxy-alpha-L-talofuranosyl)-6-thiopurine (8). Unlike the previously prepared 7-(6'-deoxy-beta-D-allofuranosyl) hypoxanthine which strongly inhibited purine nucleoside phosphorylase, compounds 6-8 did not inhibit this enzyme. Compound 8 significantly inhibited the growth of L1210 tumor cells in vitro and in vivo.