This study investigates the effect of glycyrrhetinic acid(GA) combined with doxorubicin(DOX) on apoptosis in HepG2 cells and its possible mechanisms. HepG2 cells were cultured in vitro, and cell viability was assessed using the cell counting kit-8(CCK-8) method. Flow cytometry was used to measure apoptosis levels in HepG2 cells. The cells were divided into the following groups: control group(0 μmol·L~(-1)), DOX group(2 μmol·L~(-1)), GA group(150 μmol·L~(-1)), and DOX + GA combination group(2 μmol·L~(-1) DOX + 150 μmol·L~(-1) GA), with treatments given for 24 hours. The colocalization level between the endoplasmic reticulum(ER) and mitochondria was assessed by colocalization fluorescence imaging. Fluorescence probes were used to measure the Ca~(2+) content in the ER and mitochondria. The qRT-PCR and Western blot were used to determine the mRNA and protein expression of sirtuin-3(SIRT3). Co-immunoprecipitation(CO-IP) was applied to investigate the interactions between voltage-dependent anion channel 1(VDAC1) and SIRT3, as well as between VDAC1, glucose-regulated protein 75(GRP75), and inositol 1,4,5-trisphosphate receptor(IP3R). The results showed that the combination of DOX and GA promoted apoptosis in HepG2 liver cancer cells. The colocalization level between the ER and mitochondria was significantly reduced, the Ca~(2+) content in the ER was significantly increased, and the Ca~(2+) content in the mitochondria was significantly decreased. The relative expression of VDAC1, GRP75, and IP3R was significantly reduced, and interactions between VDAC1, GRP75, and IP3R were observed. SIRT3 mRNA and protein expression levels were significantly increased, and an interaction between SIRT3 and VDAC1 was detected. The acetylation level of VDAC1 was significantly decreased. In conclusion, GA combined with DOX induces apoptosis in HepG2 cells by mediating the deacetylation of VDAC1 through SIRT3, weakening the interactions among VDAC1, GRP75, and IP3R. This regulates the formation of endoplasmic reticulum-mitochondrial membrane domains(ERMMDs), affects Ca~(2+) transport between the ER and mitochondria, and ultimately triggers cell apoptosis.
Humans ; Apoptosis/drug effects* ; Hep G2 Cells ; Glycyrrhetinic Acid/pharmacology* ; Doxorubicin/pharmacology* ; Liver Neoplasms/genetics* ; Carcinoma, Hepatocellular/physiopathology* ; Mitochondria/metabolism* ; Endoplasmic Reticulum/metabolism* ; Cell Survival/drug effects* ; Membrane Proteins/genetics*

Addressing the enduring challenge of evaluating traditional Chinese medicines (TCMs), the integrated evidence chain-based effectiveness evaluation of TCMs (Eff-iEC) has emerged. This paper explored its capacity through a demonstration study that evaluated the effectiveness evidence of six commonly used anti-hepatic fibrosis Chinese patent medicines (CPMs), including Biejiajian Pill (BP), Dahuang Zhechong Pill (DZP), Biejia Ruangan Compound (BRC), Fuzheng Huayu Capsule (FHC), Anluo Huaxian Pill (AHP), and Heluo Shugan Capsule (HSC), using both Eff-iEC and the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) system. The recognition of these CPMs within the TCM academic community was also assessed through their inclusion in relevant medical documents. Results showed that the evidence of BRC and FHC received higher assessments in both Eff-iEC and GRADE system, while the assessments for others varied. Analysis of community recognition revealed that Eff-iEC more accurately reflects the clinical value of these CPMs, exhibiting superior evaluative capabilities. By breaking through the conventional pattern of TCMs effectiveness evaluation, Eff-iEC offers a novel epistemology that better aligns with the clinical realities and reasoning of TCMs, providing a coherent methodology for clinical decision-making, new drug evaluations, and health policy formulation.

Objective To investigate the molecular mechanism of verbascoside against acute lung injury(ALI)by network pharmacology and molecular docking methods,and to validate the findings experimentally.Methods The 2D structure of verbascoside was obtained from the Pubchem database.Active ingredient targets of verbascoside were acquired from Pharmmapper database and Swiss Target Prediction database.Active component targets of ALI were acquired from datebase such as Gene Cards,OMIM,and DisGeNET.Common targets between verbascoside and ALI were determined by overlapping these sets.PPI network for potential targets was constructed using String database and Cytoscape software.The intersection targets were imported into the DAVID database for enrichment analysis of GO biological processes,KEGG signaling pathway and the pathway target genes.Molecular docking between verbascoside and core targets was performed using Autodock vina software.The mRNA expression level of core genes was validated using real-time quantitative PCR(RT-qPCR),and the expression of related proteins was detected using Western blotting.Results A total of 150 target genes of verbascoside against ALI were screened,and the key targets of verbascoside against ALI mainly involve pathways such as Rap1 signaling pathway,PI3K-Akt signaling pathway and MAPK signaling pathway.Verbascoside docked well with the core target molecules.RT-qPCR results showed that,compared with the control group,the mRNA expression levels of HSP90AA1,ALB,TP53,TNF,INS,and HRAS were significantly decreased in cells after the effect of verbascoside(P<0.05);Western blotting indicated that,compared with the model group,verbascoside treatment significantly reduced the expression of p-Akt,p-p38,and p-ERK proteins(P<0.05).Conclusion Verbascoside could inhibit MAPK,Rap1 and PI3K/Akt signaling pathways to exert its anti-ALI effects.

Objective:To analyze the influencing factors of clinically relevant postoperative pancreatic fistula (CR-POPF) in children with pancreatic tumors after surgery.Methods:The clinical data of 123 children undergoing surgery for pancreatic tumor in Beijing Children's Hospital, Capital Medical University from January 2007 to March 2020 were retrospectively analyzed, including 39 males and 84 females, with a median age of 9.8 years (6.7 to 11.8). Patients without pancreatic fistula and with biochemical leakage were included in control group ( n=95), while patients with grade B and C pancreatic fistula were divided into CR-POPF group ( n=28). The independent influencing factors of CR-POPF were analyzed by univariate and multivariate logistic regression. Results:Among 123 children, 28 cases (22.8%) developed CR-POPF, including 24 cases (85.7%, 24/28) of grade B pancreatic fistula and 4 cases (14.3%, 4/28) of grade C pancreatic fistula. There were significant differences between CR-POPF and control groups in the age > 8 years and 4 months, tumor location, operation time >390 min and procedures (all P<0.05). Multivariate logistic regression analysis showed an increased risk of CR-POPF in children aged > 8 years and 4 months ( OR=8.226, 95% CI: 1.813-37.333, P=0.006) and undergoing duodenum-preserving pancreatic head resection (DPPHR) ( OR=3.353, 95% CI: 1.282-8.767, P=0.014). Conclusion:Age>8 years and 4 months and DPPHR are independent risk factors for CR-POPF in children with pancreatic tumors after surgery.

Humans ; Forensic Medicine ; Poisoning/diagnosis*

The function of the central nervous system was significantly altered under high-altitude hypoxia, and these changes lead to central nervous system disease and affected the metabolism of drugs in vivo. The blood-brain barrier is essential for maintaining central nervous system stability and plays a key role in the regulation of drug metabolism, and barrier structure and dysfunction affect drug transport to the brain. Changes in the structure and function of the blood-brain barrier and the transport of drugs across the blood-brain barrier under high-altitude hypoxia are regulated by changes in brain microvascular endothelial cells, astrocytes and pericytes, and are regulated by drug metabolism factors such as drug transporters and drug metabolizing enzymes. This article reviews the effects of high-altitude hypoxia on the structure and function of the blood-brain barrier and the effects of changes in the blood-brain barrier on drug metabolism. We investigate the regulatory effects and underlying mechanisms of the blood-brain barrier and related pathways such as transcription factors, inflammatory factors and nuclear receptors on drug transport under high-altitude hypoxia.

Aim To analyze the molecular mechanism of rhubarb in the treatment of aeute pancreatitis ( AP) by network pharmacology and molecular docking.Methods TCMSP,TCMID and Swiss target predic¬tion databases were used to screen the active compo¬nents and targets of rhubarb,and genecards and OMIM databases were used to screen the targets of AP.Then the active ingredient drug target network of rhubarb and theactive ingredient disease target network of rhubarb for AP were constructed by using Cytoscape software.PPI network was constructed in string database, and go and KEGG enrichment analysis was performed in metascape database and R language.Finally,molecular docking was used to verify the possibility of binding the core active components to the core target.Results A total of 192 active components and 1 882 AP targets were obtained.The first three active components of rhubarb in the treatment of AP were beta sitosterol, aloe emodin and eupatin.The core target of rhubarb in the treatment of AP was hsp90aal.Go enrichment analysis focused on reaction to toxic substances, while KEGG enrichment analysis was significantly enriched in p53 signaling pathway closely related to AP.Molecular docking showed good binding and stable conformation.Conclusions Rhubarb can affect the expression of AP related genes and proteins through p53 signaling pathway, thereby inhibiting cell apoptosis and allevia¬ting the inflammatory injury of AP.

Aim To study the protective effect of Qingjie HuaGong decoction ( QJHGD) for severe acute pancreatitis ( SAP ) model rats induced by cerulein based on TLR4/NF-kB/MYD88 pathway.Methods The effective component groups and potential targets of QJHGD were collected by network pharmacology method , and we constructed the component-target network.The GO and KEGG of important targets were enriched and analyzed by metascape database, and we selected the targeted pathways related with SAP inflammation mechanism.The rat model of severe acute pancreatitis was established by cerulein combined with lipopolysac- charide, followed by QJHGD gavage.Pancreatic tissues were observed by hematoxylin and eosin staining.We verified the therapeutic effect of QJHGD on SAP rats and the regulatory effect on TLR4/NF-kB/MyD88 target pathway, by Enzyme linked immunosorbent and immunohistochemistry methods.Results A total of 105 active components and 148 key targets for SAP were screened; KEGG was enriched 320 different channels including toll like receptor and NF-kB classical pathways.Animal experiments showed that QJHGD harl protective changes in pancreatic pathological tissues, which was observed by HE staining; QJHGD reduced amylase, lipase, 1L-6 and TNF-a in SAP rat serum, inhibiting the positive expression of key proteins on TLR4/N F- kB/MyD88 inflammatory transduction j j pathways.Conclusion The mechanisms of QJHGD protecting pancreatic injury of SAP rat may be related to reducing the expression of key proteins on TLR4/ NF-kB/MvD88 pathway.

To clarify the key quality attributes of substance benchmarks in Danggui Buxue Decoction(DBD), this study prepared 21 batches of DBD substance benchmarks, and established two methods for detecting their fingerprints, followed by the identification of peak attribution and similarity range as well as the determination of extract and transfer rate ranges and contents of index components ferulic acid, calycosin-7-O-β-D-glucoside, and astragaloside Ⅳ. The mass fractions and transfer rates of DBD substance benchmarks from different batches were calculated as follows: ferulic acid(index component in Angelicae Sinensis Radix): 0.037%-0.084% and 31.41%-98.88%; astragaloside Ⅳ(index component in Astragali Radix): 0.021%-0.059% and 32.18%-118.57%; calycosin-7-O-β-D-glucoside: 0.002%-0.023% and 11.51%-45.65%, with the extract rate being 18.4%-36.1%. The similarity of fingerprints among 21 batches of DBD substance benchmarks was all higher than 0.9. The quality control method for DBD substance benchmarks was preliminarily established based on the HPLC fingerprint analysis and index component determination, which has provided a basis for the subsequent development of DBD and the quality control of novel related preparations.
Chromatography, High Pressure Liquid/methods* ; Drugs, Chinese Herbal/standards* ; Quality Control