1.Literature Based Analysis on Adverse Reactions in Simultaneously Clinical Use of Banxia (Rhizoma Pinelliae)- Wutou (Aconitum)
Can CAO ; Wenyong LIAO ; Jiwen ZHANG ; Yinghao WU ; Xiangnan XU ; Meijing WU ; Xiaoqing LIU ; Shaohong CHEN ; Haiyan LIU ; Linlin XIU ; Xiangqing CUI ; Gaoyang LI ; Ying ZHANG ; Gansheng ZHONG
Journal of Traditional Chinese Medicine 2025;66(9):955-962
ObjectiveTo analyze the adverse reactions associated with the clinical use of Banxia (Rhizoma Pinelliae)- Wutou (Aconitum) in the same formula, with the aim of providing a reference for the safety of their clinical application. MethodsLiterature on the clinical application of antagonistic herbs "Banxia-Wutou" used in the same formula, published from January 1st, 2014, to June 30th, 2023, was retrieved from databases including CNKI, VIP, Wanfang, SinoMed, PubMed, Cochrane Library, and Embase. A database was established, and information related to adverse reactions was extracted, including descriptions, classifications, specific manifestations, management and outcomes, patients' primary diseases (western medicine diseases and traditional Chinese medicine diagnoses and syndromes), and medication information (dosage, ratio, administration routes, and dosage forms). ResultsA total of 79 researches simultaneously used antagonistic herbs Banxia-Wutou in the same formula and reported associated advers reactions. Gastrointestinal adverse reactions were the most common, with 8 studies reporting management of adverse reactions and 3 studies reporting improvement with no intervention. Among the 11 researches, the adverse reaction relieved to extant, while other 69 researches didn't report the managment of adverse reaction and its prognosis. For the primary disease in western medicine system, chronic bronchitis and chronic obstructive pulmonary disease (COPD) were most common, while gastric pain was the most common symptom in traditional Chinese medicine with spleen and kidney deficiency and spleen stomach cold deficiency being the most frequent syndromes. The most common Banxia dosage was 10 g, while for the Wutou, Fuzi (Radix Aconiti Lateralis Praeparata) was predominant with the highest dose at 15 g. The most frequent herbal combination was Banxia-fuzi, with a 1∶1 ratio. The main administration route was oral, and the primary dosage form was decoction. ConclusionGastrointestinal adverse reactions are the most common in the clinical use of Banxia-Wutou antagonistic herb combinations. Research on the safety of "Banxia-Wutou" combinations should focus on respiratory system diseases and spleen-stomach related conditions.
2.Mechanisms of puerarin-mediated lipid modulation to enhance glucose-lowering effects via hepatic ChREBP/PPARα/PPARγ in vitro.
Can CUI ; Han-Yue XIAO ; Li-Ke YAN ; Zhong-Hua XU ; Wei-Hua LIU ; Hui-Ping LI ; Jun TU
China Journal of Chinese Materia Medica 2025;50(14):3951-3961
This study aims to investigate the in vitro mechanisms underlying the beneficial effects of puerarin on hepatic insulin resistance(IR) based on the carbohydrate response element-binding protein(ChREBP)/peroxisome proliferator-activated receptor(PPAR)α/PPARγ axis involved in glucose and lipid metabolism. An IR-HepG2 cell model was established by treating cells with dexamethasone for 48 h, and the cells were then treated with 10, 20, and 40 μmol·L~(-1) puerarin for 24 h. Glucose levels and output in the extracellular fluid were measured by the glucose oxidase method, while cell viability was assessed by the cell counting kit-8(CCK-8) assay. The adenosine triphosphate(ATP) content and glycogen synthesis were evaluated through chemiluminescence and periodic acid-Schiff staining, respectively. Western blot was employed to quantify the protein levels of forkhead box protein O1(FoxO1), phosphorylated forkhead box protein O1 [p-FoxO1(Ser256)], glucagon, phosphofructokinase, liver type(PFKL), pyruvate kinase L-R(PKLR), pyruvate dehydrogenase complex 1(PDHA1), insulin receptor substrate 2(IRS2), phosphatidylinositol 3-kinase p85(PI3KR1), phosphorylated protein kinase B [p-Akt(Thr308)], glycogen synthase(GYS), glycogen phosphorylase, liver type(PYGL), adiponectin(ADPN), ChREBP, PPARα, and PPARγ. Additionally, the protein levels of acetyl-CoA carboxylase 1(ACC1), phosphorylated ATP citrate lyase [p-ACLY(Ser455)], sterol regulatory element binding protein 1c(SREBP-1c), peroxisome proliferator-activated receptor gamma coactivator 1α(PGC1α), carnitine palmitoyltransferase 1α(CPT1α), and glucagon receptor(GCGR) were also determined. Immunofluorescence was employed to visualize the expression and nuclear location of ChREBP/PPARα/PPARγ. Furthermore, quantitative PCR with the antagonists GW6471 and GW9662 was employed to assess Pparα, Pparγ, and Chrebp. The findings indicated that puerarin effectively reduced both the glucose level and glucose output in the extracellular fluid of IR-HepG2 cells without obvious effect on the cell viability, and it increased intracellular glycogen and ATP levels. Puerarin down-regulated the protein levels of FoxO1 and glucagon while up-regulating the protein levels of p-FoxO1(Ser256), PFKL, PKLR, PDHA1, IRS2, PI3KR1, p-Akt(Thr308), GYS, PYGL, ADPN, ACC1, SREBP-1c, p-ACLY(Ser455), PGC1α, CPT1α, and GCGR in IR-HepG2 cells. Furthermore, puerarin up-regulated both the mRNA and protein levels of ChREBP, PPARα, and PPARγ and promoted the translocation into the nucleus. GW6471 was observed to down-regulate the expression of Pparα while up-regulating the expression of Chrebp and Pparγ. GW9662 down-regulated the expression of Pparγ while up-regulating the expression of Pparα, with no significant effect on Chrebp. In summary, puerarin activated the hepatic ChREBP/PPARα/PPARγ axis, thereby coordinating the glucose and lipid metabolism, promoting the conversion of glucose to lipids to exert the blood glucose-lowering effect.
Isoflavones/pharmacology*
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Humans
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PPAR gamma/genetics*
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Hep G2 Cells
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Glucose/metabolism*
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Lipid Metabolism/drug effects*
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PPAR alpha/genetics*
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Liver/drug effects*
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics*
;
Insulin Resistance
3.Hypoglycemic effect and mechanism of berberine in vitro based on regulation of BMAL1:CLOCK complex involved in hepatic glycolysis, glucose oxidation a nd gluconeogenesis to improve energy metabolism.
Zhong-Hua XU ; Li-Ke YAN ; Wei-Hua LIU ; Can CUI ; Han-Yue XIAO ; Hui-Ping LI ; Jun TU
China Journal of Chinese Materia Medica 2025;50(15):4293-4303
This paper aims to investigate the hypoglycemic effect and mechanism of berberine in improving energy metabolism based on the multi-pathway regulation of brain and muscle aromatic hydrocarbon receptor nuclear translocal protein 1(BMAL1): cyclin kaput complex of day-night spontaneous output cyclin kaput(CLOCK). The dexamethasone-induced hepatic insulin resistance(IR) HepG2 cell model was used; 0.5, 1, 5, 10, 20 μmol·L~(-1) berberine were administered at 15, 18, 21, 24, 30, 36 h. The time-dose effect of glucose content in extracellular fluid was detected by glucose oxidase method. The optimal dosage and time of berberine were determined for the follow-up study. Glucose oxidase method and chemiluminescence method were respectively performed to detect hepatic glucose output and relative content of ATP in cells; Ca~(2+), reactive oxygen species(ROS), mitochondrial structure and membrane potential were detected by fluorescent probes. Moreover, ultraviolet colorimetry method was used to detect the liver type of pyruvate kinase(L-PK) and phosphoenol pyruvate carboxykinase(PEPCK). In addition, pyruvate dehydrogenase E1 subunit α1(PDHA1), phosphate fructocrine-liver type(PFKL), forkhead box protein O1(FoxO1), peroxisome proliferator-activated receptor gamma co-activator 1α(PGC1α), glucose-6-phosphatase(G6Pase), glucagon, phosphorylated nuclear factor-red blood cell 2-related factor 2(p-Nrf2)(Ser40), heme oxygenase 1(HO-1), NAD(P)H quinone oxidoreductase 1(NQO1), fibroblast growth factor 21(FGF21), uncoupled protein(UCP) 1 and UCP2 were detected by Western blot. BMAL1:CLOCK complex was detected by immunofluorescence double-staining method, combined with small molecule inhibitor CLK8. Western blot was used to detect PDHA1, PFKL, FoxO1, PGC1α, G6Pase, glucagon, Nrf2, HO-1, NQO1, FGF21, UCP1 and UCP2 in the CLK8 group. The results showed that berberine downregulated the glucose content in extracellular fluid in IR-HepG2 cells in a time-and dose-dependent manner. Moreover, berberine inhibited hepatic glucose output and reduced intracellular Ca~(2+) and ROS whereas elevated JC-1 membrane potential and improved mitochondrial structure to enhance ATP production. In addition, berberine upregulated the rate-limiting enzymes such as PFKL, L-PK and PDHA1 to promote glycolysis and aerobic oxidation but also downregulated PGC1α, FoxO1, G6Pase, PEPCK and glucagon to inhibit hepatic gluconeogenesis. Berberine not only upregulated p-Nrf2(Ser40), HO-1 and NQO1 to enhance antioxidant capacity but also upregulated FGF21, UCP1 and UCP2 to promote energy metabolism. Moreover, berberine increased BMAL1, CLOCK and nuclear BMAL1:CLOCK complex whereas CLK8 reduced the nuclear BMAL1:CLOCK complex. Finally, CLK8 decreased PDHA1, PFKL, Nrf2, HO-1, NQO1, FGF21, UCP1, UCP2 and increased FoxO1, PGC1α, G6Pase and glucagon compared with the 20 μmol·L~(-1) berberine group. BMAL1:CLOCK complex inhibited gluconeogenesis, promoted glycolysis and glucose aerobic oxidation pathways, improved the reduction status within mitochondria, protected mitochondrial structure and function, increased ATP energy storage and promoted energy consumption in IR-HepG2 cells. These results suggested that berberine mediated BMAL1:CLOCK complex to coordinate the regulation of hepatic IR cells to improve energy metabolism in vitro.
Humans
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Berberine/pharmacology*
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Gluconeogenesis/drug effects*
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Hep G2 Cells
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Glucose/metabolism*
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Liver/drug effects*
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Energy Metabolism/drug effects*
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Hypoglycemic Agents/pharmacology*
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ARNTL Transcription Factors/genetics*
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Glycolysis/drug effects*
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Oxidation-Reduction/drug effects*
4.Empirical and modified hemostatic resuscitation for liver blast injury combined with seawater immersion: A preliminary study.
Haoyang YANG ; Chenglin DAI ; Dongzhaoyang ZHANG ; Can CHEN ; Zhao YE ; Xin ZHONG ; Yijun JIA ; Renqing JIANG ; Wenqiong DU ; Zhaowen ZONG
Chinese Journal of Traumatology 2025;28(3):220-225
PURPOSE:
To compare the effects of empirical and modified hemostatic resuscitation for liver blast injury combined with seawater immersion.
METHODS:
Thirty rabbits were subjected to liver blast injury combined with seawater immersion, and were then divided into 3 groups randomly (n = 10 each): group A (no treatment after immersion), group B (empirical resuscitation with 20 mL hydroxyethyl starch, 50 mg tranexamic acid, 25 IU prothrombin complex concentrate and 50 mg/kg body weight fibrinogen concentrate), and group C (modified resuscitation with additional 10 IU prothrombin complex concentrate and 20 mg/kg body weight fibrinogen concentrate based on group B). Blood samples were gathered at specified moments for assessment of thromboelastography, routine coagulation test, and biochemistry. Mean arterial pressure, heart rate, and survival rate were also documented at each time point. The Kolmogorov-Smirnov test was used to examine the normality of data distribution. Multigroup comparisons were conducted with one-way ANOVA.
RESULTS:
Liver blast injury combined with seawater immersion resulted in severe coagulo-fibrinolytic derangement as indicated by prolonged prothrombin time (s) (11.53 ± 0.98 vs. 7.61 ± 0.28, p<0.001), activated partial thromboplastin time (APTT) (s) (33.48 ± 6.66 vs. 18.23 ± 0.89, p<0.001), reaction time (R) (min) (5.85 ± 0.96 vs. 2.47 ± 0.53, p<0.001), decreased maximum amplitude (MA) (mm) (53.20 ± 5.99 vs. 74.92 ± 5.76, p<0.001) and fibrinogen concentration (g/L) (1.19 ± 0.29 vs. 1.89 ± 0.32, p = 0.003), and increased D-dimer concentration (mg/L) (0.38 ± 0.32 vs. 0.05 ± 0.03, p = 0.005). Both empirical and modified hemostatic resuscitation could improve the coagulo-fibrinolytic states and organ function, as indicated by shortened APTT and R values, decreased D-dimer concentration, increased fibrinogen concentration and MA values, lower concentration of blood urea nitrogen and creatine kinase-MB in group B and group C rabbits in comparison to that observed in group A. Further analysis found that the R values (min) (4.67 ± 0.84 vs. 3.66 ± 0.98, p = 0.038), APTT (s) (23.16 ± 2.75 vs. 18.94 ± 1.05, p = 0.001), MA (mm) (60.10 ± 4.74 vs. 70.21 ± 3.01, p < 0.001), and fibrinogen concentration (g/L) (1.68 ± 0.21 vs. 1.94 ± 0.16, p = 0.013) were remarkably improved in group C than in group B at 2 h and 4 h after injury. In addition, the concentration of blood urea nitrogen (mmol/L) (24.11 ± 1.96 vs. 21.00 ± 3.78, p = 0.047) and creatine kinase-MB (U/L) (85.50 ± 13.60 vs. 69.74 ± 8.56, p = 0.013) were lower in group C than in group B at 6 h after injury. The survival rates in group B and group C were significantly higher than those in group A at 4 h and 6 h after injury (p < 0.001), however, there were no statistical differences in survival rates between group B and group C at each time point.
CONCLUSIONS
Modified hemostatic resuscitation could improve the coagulation parameters and organ function better than empirical hemostatic resuscitation.
Animals
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Rabbits
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Resuscitation/methods*
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Liver/injuries*
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Seawater
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Blast Injuries/therapy*
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Fibrinogen/administration & dosage*
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Male
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Tranexamic Acid/administration & dosage*
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Immersion
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Hydroxyethyl Starch Derivatives/administration & dosage*
5.Laboratory Diagnosis and Molecular Epidemiological Characterization of the First Imported Case of Lassa Fever in China.
Yu Liang FENG ; Wei LI ; Ming Feng JIANG ; Hong Rong ZHONG ; Wei WU ; Lyu Bo TIAN ; Guo CHEN ; Zhen Hua CHEN ; Can LUO ; Rong Mei YUAN ; Xing Yu ZHOU ; Jian Dong LI ; Xiao Rong YANG ; Ming PAN
Biomedical and Environmental Sciences 2025;38(3):279-289
OBJECTIVE:
This study reports the first imported case of Lassa fever (LF) in China. Laboratory detection and molecular epidemiological analysis of the Lassa virus (LASV) from this case offer valuable insights for the prevention and control of LF.
METHODS:
Samples of cerebrospinal fluid (CSF), blood, urine, saliva, and environmental materials were collected from the patient and their close contacts for LASV nucleotide detection. Whole-genome sequencing was performed on positive samples to analyze the genetic characteristics of the virus.
RESULTS:
LASV was detected in the patient's CSF, blood, and urine, while all samples from close contacts and the environment tested negative. The virus belongs to the lineage IV strain and shares the highest homology with strains from Sierra Leone. The variability in the glycoprotein complex (GPC) among different strains ranged from 3.9% to 15.1%, higher than previously reported for the seven known lineages. Amino acid mutation analysis revealed multiple mutations within the GPC immunogenic epitopes, increasing strain diversity and potentially impacting immune response.
CONCLUSION
The case was confirmed through nucleotide detection, with no evidence of secondary transmission or viral spread. The LASV strain identified belongs to lineage IV, with broader GPC variability than previously reported. Mutations in the immune-related sites of GPC may affect immune responses, necessitating heightened vigilance regarding the virus.
Humans
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China/epidemiology*
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Genome, Viral
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Lassa Fever/virology*
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Lassa virus/classification*
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Molecular Epidemiology
;
Phylogeny
6.Construction of damage control operation simulation training platform for traumatic brain injury of wartime based on mixed reality
Wen-Qiong DU ; Zhao-Wen ZONG ; Xin ZHONG ; Ren-Qing JIANG ; Yi-Jun JIA ; Can CHEN ; Chuan-Shuan WANG
Chinese Medical Equipment Journal 2024;45(2):17-21
Objective To develop a damage control operation(DCO)simulation training platform for traumatic brain injury(TBI)in wartime based on mixed reality to open up a new path for surgical skills training of military surgeons.Methods The platform mainly consisted of wartime TBI DCO simulation training software,a surgical manikin and a HoloLens 2 MR device.The simulating training software was developed with C# language and the technologies of MR,basic gestures,spatial scanning positioning and etc on the basis of constructed surgical decision-making training system,virtual surgical environment and functional modules.The surgical manikin was customized with reference to the standard body type of an adult male with a height of 180 cm,and an electronic chip was developed and placed inside the head of the manikin to execute data matching with the simulation training software.The simulation training software was installed and run in the HoloLens 2 MR device to realize TBI DCO simulation training on the virtual reality interactive model.Results The platform developed implemented the functions of virtual reality interactive model reset positioning,operation simulation training,examination and on-site demonstration,which gained advantages in stimulating learning interest and facilitating risk-free,time-and space-indepen-dent,immersive and interactive learning and was generally recognized by the trainees.Conclusion The simulation training platform can be a supplementary to other training means to improve the ability of military surgeons in damage control operation.[Chinese Medical Equipment Journal,2024,45(2):17-21]
7.Development of pre-deployment base-based training course for medical units in peacekeeping level-II hospitals
Can CHEN ; Wenqiong DU ; Renqing JIANG ; Xin ZHONG ; Wei ZHANG ; Lin CHEN ; Zhaowen ZONG
Chinese Journal of Medical Education Research 2024;23(4):482-486
Objective:To establish a practical and systematic pre-deployment base-based training course for medical units in peacekeeping level-II hospitals.Methods:We surveyed the current status of pre-deployment training of medical teams in peacekeeping level-II hospitals, and organized expert discussions to preliminarily construct centralized training course contents. At the same time, the Delphi method was used to conduct two rounds of questionnaire-based consultation to evaluate the overall framework design of the course and the rationality and importance scores of each module. The contents of the pre-deployment training course were further selected and optimized based on the positive coefficient (%), authority coefficient (Cr), coefficient of concordance ( W), and coefficient of variation (Cv). Results:The contents of the pre-deployment training course for the medical units of peacekeeping level-II hospitals were preliminarily established. The experts scored high for the combat onsite first-aid skills module [rationality score, (9.32±0.75) points; importance score, (9.45±0.65) points] and the combat wound treatment module [rationality score, (9.33±0.75) points; importance score, (9.28±0.74) points], and scored lowest for the field survival module [rationality score, (7.95±2.28) points; importance score, (7.87±2.16) points].Conclusions:The established contents of the pre-deployment base-based training course for the medical units of peacekeeping level-II hospitals are reasonable and practical, which lays the foundation for perfecting the pre-deployment training system of peacekeeping medical teams.
8.Effect of Sargassum and Glycyrrhizae Radix et Rhizoma Incompatible Pair with Haizao Yuhutang on Oxidative Stress in Liver of Goiter Rats
Xiao DONG ; Yi TIAN ; Xiaoqing LIU ; Can CAO ; Wenyong LIAO ; Xiangnan XU ; Meijing WU ; Haiyan LIU ; Shaohong CHEN ; Xue YU ; Angran FAN ; Linlin XIU ; Gansheng ZHONG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):37-45
ObjectiveTo observe the effects of Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the Haizao Yuhutang (HYT) on oxidative stress in the liver of goiter rats under the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020. MethodA total of 128 male Wistar rats were randomly divided into a blank group, a model group, a euthyrox group (20 μg·kg-1), a HYT group (12.06 g·kg-1), a HYT without Sargassum (HYT-H) group (9.90 g·kg-1), a HYT without Glycyrrhizae Radix et Rhizoma (HYT-G) group (10.26 g·kg-1), a HYT without Sargassum and Glycyrrhizae Radix et Rhizoma (HYT-HG) group (8.10 g·kg-1), and a Sargassum and Glycyrrhizae Radix et Rhizoma (HG) group (3.96 g·kg-1). The blank group was given deionized water by gavage, and the others were given propylthiouracil (PTU) to replicate the goiter pathological model. Euthyrox was taken as a positive control drug, and the rest of the Chinese medicine groups were given the corresponding decoction by gavage, the material was collected 12 hours after the last dose. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), reactive oxygen species (ROS) in liver tissue were detected in each group. The pathological changes in the liver were observed via hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was utilized to detect the mRNA expressions of Kelch-like Ech-associated protein 1 (Keap1), nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), p53 and Caspase-3 in liver tissues. Western blot was adopted to detect the protein expressions of Nrf2 and HO-1 in liver tissues in oxidative stress-related signaling pathways. ResultCompared with control group, the model group showed significantly increased serum ALT level and contents of MDA and ROS in liver tissues (P<0.05, P<0.01), significantly reduced activities of SOD and GSH-Px in the liver (P<0.01), significantly increased mRNA expression of Keap1 (P<0.01), and significantly decreased mRNA and protein expressions of Nrf2 and HO-1 (P<0.05, P<0.01). Compared with the model group, the HYT group manifested significantly reduced serum levels of AST, ALT, and ALP (P<0.05, P<0.01), significantly reduced contents of MDA and ROS in liver tissue (P<0.01), significantly increased the activities of SOD and GSH-Px (P<0.01), significantly decreased mRNA expressions of Keap1, p53, and Caspase-3 (P<0.01), and significantly increased mRNA and protein expressions of Nrf2 and HO-1 (P<0.05, P<0.01). ConclusionUnder the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020, Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the HYT on oxidative stress in the liver of goiter rats had different effects. The HYT that contains Sargassum and Glycyrrhizae Radix et Rhizoma has a protective effect on the liver of goiter rats, and the effect is better than that of the HG group, the euthyrox group, and the incomplete groups. Its mechanism may be related to activating the Nrf2/HO-1 signaling pathway to alleviate liver oxidative stress and inhibiting the p53/Caspase-3 signaling pathway to reduce hepatocyte apoptosis.
9.Berberine regulates glucose and lipid metabolism via clock-controlled genes to ameliorate insulin resistance of hepatocytes.
Li-Ke YAN ; Can CUI ; Ying WANG ; Shui-Lan ZHU ; Zhong-Hua XU ; Han-Yue XIAO ; Wei-Hua LIU ; Jun TU
China Journal of Chinese Materia Medica 2024;49(23):6368-6377
This study aims to investigate the mechanism of berberine in regulating the metabolism network via clock-controlled genes represented by brain and muscle arnt-like 1(BMAL1) to ameliorate insulin resistance(IR) of hepatocytes in vitro. The HepG2 cell model of dexamethasone-induced IR(IR-HepG2) was established and treated with 5, 10, and 20 μmol·L~(-1) berberine, respectively, for 24 h. The glucose oxidase method and cell counting kit-8(CCK-8) assay were employed to measure extracellular glucose concentration and cell viability, respectively. Periodic acid-Schiff(PAS) staining and lipid fluorescence method were used to detect glycogen and lipids. The immunofluorescence(IF) assay was employed to detect the nuclear localization of BMAL1 and circadian locomotor output cycles kaput(CLOCK) in IR-HepG2 cells. Western blot was employed to determine the protein levels of BMAL1, CLOCK, period circadian clock 2(PER2), cryptochrome circadian regulator 1(CRY1), Rev-Erbα, carbohydrate response element-binding protein(ChREBP), peroxisome proliferator-activated receptors alpha and gamma(PPARα/γ), sterol regulatory element-binding protein 1C(SREBP-1C), mammalian target of rapamycin(mTOR), protein kinase B(Akt), glycogen synthase kinase-3β(GSK3β), acetyl coenzyme A carboxylase 1(ACC1), fatty acid synthase(FASN), carnitine palmitoyltransferase 1α(CPT1α), nicotinamide phosphoribosyltransferase(NAMPT), silent information regulator 1(SIRT1), adiponectin(ADPN), insulin receptor substrate 2(IRS2), and phosphatidylinositol 3-kinase regulatory subunit p85(PI3Kp85). In addition, the levels of phosphorylated adenosine monophosphate-activated protein kinase alpha(AMPKα), Akt, GSK3β, BMAL1, and mTOR were determined. Furthermore, 20 μmol·L~(-1) CLK8 was added to measure the glucose consumption as well as the protein levels of ChREBP, PPARα, and mTOR in IR-HepG2 cells. The results showed that berberine increased the glucose consumption, lowered the lipid levels, increased the expression and nuclear localization of BMAL1 and CLOCK, and up-regulated the level of BMAL1 in IR-HepG2 cells. Furthermore, berberine up-regulated the levels of ADPN, IRS2, PI3Kp85, p-Akt(Ser473)/Akt, p-mTOR(Ser2448)/mTOR, PPARα, and CPT1α, and down-regulated the levels of p-GSK3β(Ser9)/GSK3β, ChREBP, SREBP-1C, ACC1, and FASN. The addition of CLK8 reduced glucose consumption in IR-HepG2 cells, up-regulated the ChREBP level, and down-regulated PPARα and mTOR levels by inhibiting the BMAL1 and CLOCK interaction. In summary, berberine regulated glucose and lipid metabolism via clock-controlled genes with BMAL1 at the core to ameliorate IR of hepatocytes.
Humans
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Hepatocytes/drug effects*
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Lipid Metabolism/drug effects*
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Glucose/metabolism*
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Berberine/pharmacology*
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Insulin Resistance
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Hep G2 Cells
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CLOCK Proteins/genetics*
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ARNTL Transcription Factors/genetics*
10.Protective Effect of Flos Puerariae,Hoveniae Semen, and Their Compatibility on Acute Alcoholic Gastric Mucosal Injury in Mice and Mechanism
Wenyong LIAO ; Muyun LI ; Xiao DONG ; Can CAO ; Min HUO ; Yuna REN ; Haiyan LIU ; Linlin XIU ; Cheng HE ; Angran FAN ; Shaohong CHEN ; Gansheng ZHONG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):39-47
ObjectiveTo explore the improvement effect of Flos Puerariae, Hoveniae Semen, and their compatibility on acute alcoholic gastric mucosal injury, and lay a foundation for further development of Flos Puerariae, Hoveniae Semen, and their compatibility in the prevention and treatment of alcohol-induced multiple organ injury. MethodThe acute alcohol-induced gastric mucosal injury model of mice was established by multiple intragastric administration of 56% Hongxing Erguotou liquor (15 mL·kg-1). A total of 120 male ICR mice were randomly divided into 8 groups, namely, the blank group, model group, omeprazole group (0.026 g·kg-1), Flos Puerariae-Hoveniae Semen (compatibility) high, medium, and low-dose groups (29.2,14.6, 7.3 g·kg-1), Flos Puerariae group (19.5 g·kg-1), and Hoveniae Semen group (19.5 g·kg-1), with 15 mice in each group. After one week of adaptive feeding, the animals were pre-administrated with the corresponding drug at the rate of 10 mL·kg-1 for 3 d. From the 4th day, after 1 h of administration, Erguotou liquid was administrated at the rate of 15 mL·kg-1 and the blank group was administrated with the same volume of deionized water to record the drunkenness and sober up time. The administration was lasted for 3 d. One hour after the last administration, the eyeballs were removed and the mice were sacrificed. The concentration of ethanol in serum was determined by gas chromatograph, and the activity of ethanol dehydrogenase (ADH) in gastric mucosa was determined by ultraviolet-vis spectrophotometer. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in gastric mucosa. Serum inflammatory factors were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of nuclear transcription factor-κB (NF-κB) p65 and NF-κB inhibitory protein α (IκBα) were detected by real-time polymerase chain reaction (Real-time PCR). ResultAs compared with the normal group, the content of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in serum of mice in the model group was increased (P<0.05), the mRNA expression of NF-κB p65 in gastric mucosa tissues was increased (P<0.01), and the mRNA expression of IκBα was decreased (P<0.01). As compared with the model group, the drunkenness time of the omeprazole group, high and medium-dose compatibility groups, and Flos Puerariae group was prolonged (P<0.05), the sober up time of the high and medium-dose compatibility groups was shortened (P<0.05), the ethanol concentration in the serum of the high-dose compatibility group was decreased (P<0.05), the ADH activity in the gastric mucosa of the omeprazole group and high and medium-dose compatibility groups was increased (P<0.05), the macroscopic injury score of the high, medium, and low-dose compatibility groups and Flos Puerariae group was decreased (P<0.05), the score of pathological injury in the omeprazole group, high, medium, and low-dose compatibility groups, and Flos Puerariae group was decreased (P<0.01), the expression of IL-6 in serum of all drug groups was decreased (P<0.05), the expression of IL-1β in serum of the omeprazole group, high, medium, and low-dose Flos Puerariae groups, and Hoveniae Semen group was decreased (P<0.05), the expression of TNF-α in serum of high and medium-dose groups was decreased (P<0.05), the mRNA expression of NF-κB p65 in gastric mucosa tissues of all drug groups was decreased (P<0.05), and the mRNA expression of IκBα in gastric mucosa tissues of the omeprazole group and high, medium, and low-dose compatibility groups was increased (P<0.05). As compared with the high-dose compatibility group, the drunkenness time in the low-dose compatibility group and Hoveniae Semen group was shortened (P<0.01), the sober up time in the Flos Puerariae and Hoveniae Semen groups was prolonged (P<0.01), the concentration of ethanol in the serum of the medium and low-dose compatibility groups, Flos Puerariae group, and Hoveniae Semen group increased (P<0.05), the macroscopic injury score of the medium and low-dose compatibility groups and Hoveniae Semen group was increased (P<0.05), the pathological injury score of the medium and low-dose compatibility groups, Flos Puerariae group, and Hoveniae Semen group was increased (P<0.01), the content of IL-1β in serum of low-dose compatibility group, Flos Puerariae group, and Hoveniae Semen group was increased (P<0.01), and the mRNA expression of IκBα in gastric mucosa of the Flos Puerariae group and Hoveniae Semen group was decreased (P<0.05). As compared with the medium-dose compatibility group, the drunkenness time in the Hoveniae Semen group was shortened (P<0.05), the sober up time in the Flos Puerariae group was prolonged (P<0.05), the pathological injury score in the Flos Puerariae group and Hoveniae Semen group was increased (P<0.01), and the content of IL-1β in serum of the low-dose compatibility group, the Flos Puerariae group, and Hoveniae Semen group was increased (P<0.05). As compared with the low-dose compatibility group, the pathological injury score of the Hoveniae Semen group was increased (P<0.05). ConclusionFlos Puerariae, Hoveniae Semen, and their compatibility play a role in preventing and treating acute alcoholic gastric mucosal injury in mice, which may be related to the inhibition of the expression of NF-κB signal pathway in gastric mucosa, and the high-dose compatibility group has the optimal effect.

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