Introduction: The gut microbiota plays an important role in the development of nonalcoholic steatohepatitis (NASH). This study investigated the preventive effect of Dendrobium officinale (DO), including whether its effect was related to the gut microbiota, intestinal permeability and liver inflammation.Methods: A NASH model was established in rats using a high-fat diet (HFD) and gavage with different doses of DO or Atorvastatin Calcium (AT) for 10 weeks. Body weight and body mass index along with liver appearance, weight, index, pathology, and biochemistry were measured to assess the preventive effects of DO on NASH rats. Changes in the gut microbiota were analyzed by 16S rRNA sequencing, and intestinal permeability and liver inflammation were determined to explore the mechanism by which DO treatment prevented NASH.Results: Pathological and biochemical indexes showed that DO was able to protect rats against HFD-induced hepatic steatosis and inflammation. Results of 16S rRNA sequencing showed that Proteobacteria, Romboutsia, Turicibacter, Lachnoclostridium, Blautia, Ruminococcus_torques_group, Sutterella, Escherichia-Shigella, Prevotella, Alistipes, and Lactobacillus_acidophilus differed significantly at the phylum, genus, and species levels. DO treatment modulated the diversity, richness, and evenness of gut microbiota, downregulated the abundance of the Gram-negative bacteria Proteobacteria, Sutterella, and Escherichia-Shigella, and reduced gut-derived lipopolysaccharide (LPS) levels. DO also restored expression of the tight junction proteins, zona occludens-1 (ZO-1), claudin-1, and occludin in the intestine and ameliorated the increased intestinal permeability caused by HFD, gut microbiota such as Turicibacter, Ruminococcus, Escherichia-Shigella, and Sutterella, and LPS. Lower intestinal permeability reduced LPS delivery to the liver, thus inhibiting TLR4 expression and nuclear factor-kappaB (NF-κB) nuclear translocation, improving liver inflammation.Discussion: These results suggest that DO may alleviate NASH by regulating the gut microbiota, intestinal permeability, and liver inflammation.

Tanshinone IIA (TIIA) is a major component extracted from the traditional herbal medicine Salvia miltiorrhiza and has been indicated to play a role in the treatment of organ fibrosis. However, the evidence supporting its antifibrotic effect is insufficient and the underlying mechanism is unclear. To investigate the therapeutic effect of TIIA on non‑alcoholic steatohepatitis‑related fibrosis (NASH‑F), the present study used a methionine choline deficiency diet to induce NASH‑F in rats, and explored the effect of TIIA on the transforming growth factor‑β1 (TGF‑β1)/Smad signaling pathway. Wistar rats were randomly divided into control, NASH‑F and TIIA groups. After 8 weeks of treatment, the levels of serum markers associated with liver function and fibrosis were measured, liver fat vacuoles and inflammation were assessed by haematoxylin and eosin staining, and liver fibrosis was assessed by Masson's trichrome staining. TGF‑β1, Smad2, Smad3, Smad7 and α‑smooth muscle actin (α‑SMA) mRNA expression, and TGF‑β1, Smad2/3, phosphorylated (p)‑Smad2/3, Smad7 and α‑SMA protein levels were determined. The results revealed that TIIA could remarkably ameliorate liver fat vacuoles and inflammation in NASH‑F rats, and could decrease the levels of serum aspartate aminotransferase, alanine aminotransferase, total bilirubin, total bile acid, hyaluronic acid, type Ⅳ collagen, laminin and type III collagen, while increasing the levels of total cholesterol and triglycerides; however, this was not statistically significance. TIIA markedly suppressed the increased TGF‑β1, Smad2, Smad3 and α‑SMA mRNA expression levels observed in the liver of NASH‑F rats, while it increased the mRNA expression level of Smad7. Similarly, TIIA suppressed the increased TGF‑β1, p‑Smad2/3 and α‑SMA protein levels observed in the liver of NASH‑F rats, while it increased the protein expression level of Smad7 in vitro and in vivo. TIIA had no significant cytotoxic effect at 10, 20, 40 and 80 µmol/l on human LX‑2 cell. In conclusion, the findings of the present study indicated that TIIA alleviated NASH‑F by regulating the TGF‑β1/Smad signaling pathway. TIIA may be a useful tool in the prevention and treatment of NASH‑F.

Background: Compared with random copolymers, block copolymerization is easier to prepare for nanoparticles with core-shell structure, and they will have better glucose sensitivity and higher insulin loading. Purpose: In our study, insulin-loaded poly (3-acrylamidophenylboronic acid-block-N-vinyl caprolactam) p(AAPBA-b-NVCL) nanoparticles were successfully prepared and were glucose-sensitive, which could effectively lower the blood sugar levels within 72 hrs. Methods: The polymer of p(AAPBA-b-NVCL) was produced by reversible addition-fragmentation chain transfer polymerization based on different ratios of 3-acrylamidophenylboronic acid (AAPBA) and N-vinylcaprolactam (NVCL), and its structure was discussed by Fourier transform infrared spectroscopy and 1H-nuclear magnetic resonance . Next, the polymer was manufactured into the nanoparticles, and the characteristics of nanoparticles were detected by dynamic light scattering, lower critical solution temperature, and transmission electron microscopy. After that, the cell and animal toxicity of nanoparticles were also investigated. Results: The results demonstrated that p(AAPBA-b-NVCL) was successfully synthesized, and can be easily self-assembled to form nanoparticles. The new nanoparticles included monodisperse submicron particles, with the size of the nanoparticle ranged between 150 and 300nm and are glucose- and temperature-sensitive. Meanwhile, insulin can be easily loaded by p(AAPBA-b-NVCL) nanoparticles and an effective sustained release of insulin was observed when the nanoparticles were placed in physiological saline. Besides, MTT assay revealed that cell viability was more than 80%, and mice demonstrated no negative impact on blood biochemistry and heart, liver, spleen, lung, and kidney after intraperitoneal injection of 10 mg/kg/d of nanoparticles. This suggested that the nanoparticles were low-toxic to both cells and animals. Moreover, they could lower the blood sugar level within 72h. Conclusion: Our research suggested that these p(AAPBA-b-NVCL) nanoparticles might have the potential to be applied in a delivery system for insulin or other hypoglycemic proteins.

As a traditional Chinese medicine, Salvia miltiorrhiza Bunge was first recorded in the Shennong Materia Medica Classic and is widely used to treat “the accumulation of symptoms and masses”. The main active ingredient of Salvia miltiorrhiza Bunge, Tanshinone IIA (TIIA), has shown anti-inflammatory, antitumor, antifibrosis, antibacterial, and antioxidative activities, etc. In this study, the results showed that TIIA could inhibit the proliferation and migration of HepG2 cells and downregulate glutathione (GSH) and Glutathione Peroxidase 4 (GPX4) levels; besides, TIIA induced the production of Reactive Oxygen Species (ROS), and upregulated the total iron content. Based on network pharmacology analysis, the antitumor effect of TIIA was found to be focused on the endoplasmic reticulum (ER)-mediated ferroptosis signaling pathway, with protein kinase R (PKR)-like ER kinase (PERK)-activating transcription factor 4 (ATF4)-heat shock 70 kDa protein 5 (HSPA5) as the main pathway. Herein, TIIA showed typical ferroptosis characteristics, and a ferroptosis inhibitor (ferrostatin-1) was used to verify the effect. The antitumor effects of TIIA, occurring through the inhibition of the PERK-ATF4-HSPA5 pathway, were further observed in vivo as significantly inhibited tumor growth and the improved pathological morphology of tumor tissue in H22-bearing mice. In summary, the antitumor mechanism of TIIA might be related to the downregulation of the activation of PERK-ATF4-HSPA5 pathway-mediated ferroptosis.

The Na⁺/K⁺-ATPase α1 subunit (ATP1A1) is a potential target for hepatic carcinoma (HCC) treatment, which plays a key role in Na⁺/K⁺ exchange, metabolism, signal transduction, etc. In vivo, we found that Panax notoginseng saponins (PNS) could inhibit tumor growth and significantly downregulate the expression and phosphorylation of ATP1A1/AKT/ERK in tumor-bearing mice. Our study aims to explore the potential effects of PNS on the regulation of ATP1A1 and the possible mechanisms of antitumor activity. The effects of PNS on HepG2 cell viability, migration, and apoptosis were examined in vitro. Fluorescence, Western blot, and RT-PCR analyses were used to examine the protein and gene expression. Further analysis was assessed with a Na⁺/K⁺-ATPase inhibitor (digitonin) and sorafenib in vitro. We found that the ATP1A1 expression was markedly higher in HepG2 cells than in L02 cells and PNS exhibited a dose-dependent effect on the expression of ATP1A and the regulation of AKT/ERK signaling pathways. Digitonin did not affect the expression of ATP1A1 but attenuated the effects of PNS on the regulation of ATP1A1/AKT/ERK signaling pathways and enhanced the antitumor effect of PNS by promoting nuclear fragmentation. Taken together, PNS inhibited the proliferation of HepG2 cells via downregulation of ATP1A1 and signal transduction. Our findings will aid a data basis for the clinical use of PNS.

Background: Non-alcoholic steatohepatitis (NASH) is a progressive form of non-alcoholic fatty liver disease (NAFLD) characterised by high prevalence, increasing incidence among younger individuals and poor prognosis. NASH pathophysiology is not completely understood, and at present, there are no viable pharmacological treatments for this condition in clinical practice. Gentiopicroside (GPS). Can alleviate NASH by reducing inflammatory responses, inhibiting oxidative stress and influencing blood lipid levels. Objective: To examine the preventive and therapeutic effects of gentiopicroside (GPS) on proteins and metabolites related to the endogenous formaldehyde-homocysteine (FA-HCY) pathway as well as on oxidative stress in NASH rats. Methods: A high-fat, high-sugar diet was used to create a NASH rat model, and the rats’ liver weight, body weight and liver index levels were measured. Oil red O and hematoxylin and eosin (HE) stainings were used to detect pathological alterations in the rat livers. Serum biochemical kits were used to identify biochemical markers in the rat serum. Markers linked to oxidative stress and metabolites associated with the endogenous FA-HCY pathway index were identified using Enzyme-linked immunosorbent assays (ELISA) kits. Gas chromatography was employed to measure the amount of endogenous FA in the liver and serum. Western blotting and real-time (RT) PCR were used to determine the relative expression levels of proteins and mRNAs associated with the endogenous FA-HCY pathway in rat liver tissues. Immunofluorescence was used to measure the relative fluorescence intensities of the proteins MTHFR, MAT1A and ALDH2. Results: A diet rich in fats and sugars results in weight gain and considerable steatosis. GPS can mitigate hepatic steatosis in NASH rats, reduce NAFLD activity scores and decrease the oil red O-stained area. The NASH rats’ serum biochemical markers should be improved. In groups treated with GPS (low, medium and high dosages), the serum exhibited increased HDL-C levels ( P < 0.05) and considerably lowered the levels of total cholesterol, TG, ALT, AST and LDL-C. According to oxidative stress indicators, GPS increased SOD, GSH, Glutathione S-transferases (GST) and CAT levels ( P < 0.05) and decreased MDA and reactive oxygen species levels in the blood of NASH rats. Serum pathway indicators demonstrated that GPS also decreased the levels of SAH and HCY in the serum and endogenous FA in the livers of NASH rats ( P < 0.05) while simultaneously increasing the level of SAM ( P < 0.05). Compared with the model group, the medium-dose GPS treatment group exhibited elevated endogenous FA levels in the blood ( P < 0.05). Protein analysis revealed a significant reduction in AHCY expression ( P < 0.05) but a significant increase in ALDH2 and CBS levels ( P < 0.05) in the GPS treatment groups. RT PCR results indicated that GPS decreased AHCY and related mRNA expression ( P < 0.05) while enhancing the levels of MTHFR, MAT1A and ALDH2 and related mRNA expression ( P < 0.05). Conclusion: The GPS has therapeutic benefits in NASH rats as it improves liver weight, body weight, liver index levels, liver steatosis and liver function. The efficacy of GPS in ameliorating NASH in rats may be associated with the modulation of the endogenous FA-HCY pathway and attenuation of oxidative stress.