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Objective:To investigate the effect of total saponins of Panax japonicus(TSPJ) on ferroptosis of myocardial cells in diabetic cardiomyopathy(DCM) rats and underlying mechanism.Methods:Experiment 1: SD rats were divided into control group, DCM group, low-dose TSPJ group, high-dose TSPJ group, and metformin(Met) group, with 10 rats in each group. Experiment 2: SD rats were divided into control group, DCM group, TSPJ group, adenosine monophosphate-activated protein kinase(AMPK) inhibitor Compound C group, and TSPJ+ AMPK agonist AICAR group, with 10 rats in each group. Except for the control group, all rats were intraperitoneally injected with streptozotocin to construct a DCM model. After 8 weeks of corresponding drug intervention, the body weight as well as glucose and lipid metabolism of rats in each experimental group were assessed, and the cardiac function indicators were detected with echocardiography. The levels of serum lactate dehydrogenase(LDH), cardiac troponin I(cTnI) and creatine kinase isoenzyme MB(CK-MB) were detected by ELISA technique. The pathological changes of myocardial tissue were observed using hematoxylin-eosin(HE) staining. The levels of dismutase(SOD), glutathione(GSH), malondialdehyde(MDA), reactive oxygen species(ROS) and Fe 2+ in myocardial tissue were detected. Western blot was used to detect ferroptosis, ferritinophagy, and the AMPK/mammalian target of rapamycin/UNC-51-like kinase 1(mTOR/ULK1) signaling pathway related proteins expression in myocardial tissue. Results:Compared with control group, left ventricular ejection fraction(EF), left ventricular short axis shortening rate(FS), peak blood velocity ratio(E/A) between early and late diastolic periods were significantly decreased in DCM group, left ventricular inner diameter(LVEDd) was increased, and the serum LDH, cTnI, CK-MB were increased, the levels of SOD, GSH were decreased, MDA, ROS, Fe 2+ were increased in myocardial tissue, the expressions of TFR1, NCOA4 LC3-II/LC3-I, Beclin-1, phosphorylated AMPK and phosphorylated ULK1 were increased, the expressions of GPX4, SLC7A11 and phosphorylated mTOR were decreased. Compared with DCM group, the above indicators of rats were significantly improved in each treatment group. Compared with the TSPJ group, the AMPK agonist AICAR reversed the effects of TSPJ on ferroptosis and ferritinophagy mediated by the AMPK/mTOR/ULK1 pathway in DCM rat cardiomyocytes. Conclusion:TSPJ can inhibit ferroptosis in DCM rat cardiomyocytes and improve myocardial injury by regulating AMPK/mTOR/ULK1 mediated ferritinophagy.
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Objective To observe the effects of total saponins of Trillium tschonoskii Maxim(TST)on vascular cognitive impairment(VCI),neurovascular units(NVUs),and neural circuit integrity in rats.Methods Forty-eight male Sprague-Dawley(SD)rats were randomly divided into sham-operated group,model group,TST group(intragastric administration,100 mg/kg),and donepezil group(intragastric administration,0.45 mg/kg),and then subjected to ischemic stroke by 2-VO method(bilateral common carotid artery ligation)or sham surgery.After 28 days of intragastric administration,Mirros water maze test was performed to evaluate the spatial learning and memory abilities of rats in each group.HE and Nissl staining were used to observe the pathological changes of brain tissue in rats.The expression of synuclein(SYN)in rat hippocampus was observed by immunohistochemical staining.Changes in dendritic spines in rat's hippocampal neurons were observed by Golgi staining.Western blotting was used to detect the expression levels of IL-1β,IL-10,vascular endothelial growth factor A(VEGFA),postsynaptic density protein 95(PSD95),and growth associated protein 43(GAP43)in rat's hippocampus in each group.Results In Mirros water maze test,rats in model group showed significant prolonged escape latency(P<0.05),and a significant reduction in the number of crossing platforms and the percentage of activity time in the target quadrant(P<0.05)than those in sham-operated group;while rats in TST group and donepezil group showed significant shortened escape latency(P<0.01),and significant increase of the number of times of crossing platforms and the percentage of activity time in the target quadrant(P<0.05)than those in model group.Compared of sham-operated group,model group showed a decrease in the expression of SYN and the number of neurons,Nissl bodies,and dendritic spines in the CA1 region of the hippocampus(P<0.05).Compared with model group,TST group and donepezil group showed an increase in the expression of SYN and the number of neurons,Nissl bodies,and dendritic spines in the CA1 region of the hippocampus(P<0.05).Western blotting showed a significant increase in the expression of IL-1β and VEGF(P<0.05),and a decrease in the expression of IL-10,PSD95,and GAP43(P<0.01)in rat's hippocampus of model group than those in sham-operated group.Compared with model group,TST group and donepezil group showed a significant decrease in the expression of IL-1β(P<0.05),and an increase in the expression of VEGFA,IL-10,and GAP43(P<0.05).Conclusions TST could alleviate cognitive impairment through promoting synaptic plasticity and neurovascular unit remodeling in 2-VO model rats,suggesting its significance as a potential drug for apoplexy.
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Paridis Rhizoma possesses the functions of clearing heat and detoxifying, alleviating swelling and relieving pain, cooling the liver and calming the convulsion. Saponins are the main active components of Paridis Rhizoma. Studies have shown that total saponins in Paridis Rhizoma have obvious inhibitory effect on solid tumors such as breast cancer, lung cancer, gastric cancer, and liver cancer and non-solid tumors such as leukemia. The saponins may exert the anti-tumor effects by inhibiting the proliferation, migration, and invasion of tumor cells, regulating cell cycle, inducing apoptotic and non-apoptotic death pathways, and regulating metabolism and tumor microenvironment. Furthermore, total saponins in Paridis Rhizoma showed anti-inflammatory, antioxidant, antimicrobial, hemostatic, and uterus-contracting activities. At the same time, they may induce apoptosis of normal cells, inflammation and oxidative stress, and metabolic disorders. In recent years, the reports of liver injury, reproductive injury, gastrointestinal injury, hemolysis, and other adverse reactions caused by total saponins in Paridis Rhizoma have been increasing. Pharmacokinetic studies have shown that there are significant differences in the metabolism of total saponins in Paridis Rhizoma administrated in different ways. Injection has a fast clearance rate, while oral administration may have hepatoenteric circulation. Meanwhile, due to the low solubility and activation of P-glycoprotein (P-gp) molecular pump, the prototype absorption, intestinal permeability, and recovery rate of total saponins in Paridis Rhizoma are poor, which affects the bioavailability. The bioavailability can be improved to some extent by preparing new dosage forms or new drug delivery systems with advanced technology. This paper reviews the pharmacological effect, pharmacokinetics, and adverse reactions of Rhizoma Paridis total saponins by searching the China National Knowledge Infrastructure (CNKI), VIP, and Web of Science with ''Rhizoma Paridis total saponins'' as the keywords, hoping to provide references for the research, development, and clinical application of such components.
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ObjectiveTo investigate the role and mechanism of total saponins of Dioscorea (TSD) in mitigating nonalcoholic steatohepatitis (NASH) in mice. MethodForty-eight C57BL/6J mice were randomized into a normal group and a modeling group. The mice for modeling were fed with a high-fat and high-cholesterol diet + 20% fructose solution for 16 weeks and randomized into model, atorvastatin (4 mg·kg-1·d-1), and high-, medium-, and low-dose (200, 60, and 20 mg·kg-1·d-1) TSD groups. The mice were administrated with corresponding doses of drugs by gavage for 8 weeks. The mouse activity, liver index, levels of total cholesterol (TC), triglycerides (TG), and free fatty acids (FFAs) in the liver, and levels of TC, TG, aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transferase (GGT), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the serum were measured. Hematoxylin-eosin staining, Masson staining, oil red O staining, and transmission electron microscopy were employed to observe the pathological changes, lipid accumulation, and morphological changes of liver ultrastructure. Western blot was employed to determine the protein levels of AMP-activated protein kinase (AMPK), sterol regulatory element-binding protein-1c (SREBP-1c), acetyl-CoA carboxylase (ACC), and phosphorylated ACC (p-ACC) in the liver tissue. ResultCompared with the normal group, the activity of mice in the model group decreased(P<0.05, P<0.01), the levels of TC, TG, FFA and serum TC, TG, ALT, AST, GGT, IL-1β and TNF-α, liver coefficient and liver pathology scores were significantly increased, the expression of p-AMPK/AMPK and p-ACC proteins in liver tissues was significantly reduced, and the expressions of SREBP-1c and ACC proteins were significantly increased (P<0.01). Compared with the model group, atorvastatin increased the mouse activity (P<0.05), while each dose of TSD caused no significant changed in the mouse activity. The levels of TC, TG, FFA in liver and serum TC, TG, ALT, AST, GGT, IL-1β, TNF-α, liver coefficient and liver pathological score in TSD and atorvastatin groups were significantly decreased, and the expressions of p-AMPK/AMPK and p-ACC in liver tissue were significantly increased. The expressions of SREBP-1c and ACC were significantly decreased (P<0.05,P<0.01). ConclusionTSD may alleviate NASH in mice by regulating the AMPK/SREBP-1c/ACC signaling pathway to reduce lipid synthesis.
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ObjectiveTo investigate the effect and mechanism of total saponins from Panax japonicus (TSPJ) on white adipose tissue (WAT) browning/brown adipose tissue (BAT) activation in C57BL6/J male mice fed on a high-fat diet (HFD). MethodThirty-two C57BL6/J male mice (8-week-old) were randomly divided into a normal group, a model group, a low-dose TSPJ group, and a high-dose TSPJ group. The mice in the low-dose and high-dose TSPJ groups were given TSPJ for four months by gavage at 25, 75 mg·kg-1·d-1, respectively, and those in the other groups were given 0.5% sodium carboxymethyl cellulose (CMC-Na) accordingly. After four months of feeding, all mice were placed at 4 ℃ for acute cold exposure, and the core body temperature was monitored. Subsequently, all mice were sacrificed, and BAT and inguinal WAT (iWAT) were separated rapidly to detect the corresponding indexes. Hematoxylin-eosin (HE) staining was used to observe the morphological changes in each group. The effect of TSPJ on the mRNA expression of uncoupling protein 1 (UCP1), fatty acid-binding protein 4 (FABP4), cytochrome C (CytC), PR domain-containing protein 16 (PRDM16), elongation of very long chain fatty acids protein 3 (ELOVL3), peroxisome proliferator-activated receptor γ (PPARγ), and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) in iWAT and BAT was detected by Real-time polymerase chain reaction (Real-time PCR). Western blot was also used to detect the protein expression of UCP1, PRDM16, PPARγ, and PGC-1α in BAT and iWAT of each group. The effect of TSPJ on UCP1 expression in BAT and iWAT was detected by immunohistochemistry. Result① Compared with the model group, TSPJ could decrease the body weight and proportions of iWAT and BAT in the HFD-induced mice (P<0.05, P<0.01). ② The body temperature of mice in the model group decreased compared with that in the normal group in the acute cold exposure tolerance test (P<0.05). The body temperature in the high-dose TSPJ group increased compared with that in the model group (P<0.01). ③ Compared with the normal group, the model group showed increased adipocyte diameter in iWAT and BAT and decreased number of adipocytes per unit area. Compared with the model group, the TSPJ groups showed significantly reduced cell diameter and increased number of cells per unit area, especially in the high-dose TSPJ group. ④ Compared with the normal group, the model group showed decreased mRNA expression of FABP4, UCP1, CytC, PRDM16, ELOVL3, PGC-1α, and PPARγ in adipose tissues of mice (P<0.05, P<0.01). Compared with the model group, after intervention with TSPJ, the mRNA expression was significantly up-regulated (P<0.05, P<0.01). ⑤ Compared with the normal group, the model group showed decreased protein expression of UCP1, PRDM16, PPARγ, and PGC-1α in adipose tissues of mice (P<0.05, P<0.01). Compared with the model group, after intervention with TSPJ, the protein expression increased significantly (P<0.05, P<0.01). ConclusionTSPJ could induce the browning of iWAT/BAT activation and enhance adaptive thermogenesis in obese mice induced by HFD. The underlying mechanism may be attributed to the activation of the PPARγ/PGC-1α signaling pathway.
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With the improvement of living standards and changes in working style, the prevalence of abnormal glucose and lipid metabolism in humans is increasing in modern society. Clinically, the related indicators are often improved by changing the lifestyle and/or taking hypoglycemic and lipid-lowering drugs, but there are no therapeutic drugs for disorders of glucose and lipid metabolism at present. Hepatitis C virus core protein binding protein 6(HCBP6) is a newly discovered target that can regulate triglyceride and cholesterol content according to level oscillations in the body, thereby regulating abnormal glucose and lipid metabolism. Relevant studies have shown that ginsenoside Rh_2 can significantly up-regulate the expression of HCBP6, but there are few studies on the effect of Chinese herbal medicines on HCBP6. Moreover, the three-dimensional structural information of HCBP6 has not been determined and the discovery of potential active components acting on HCBP6 is not rapidly advanced. Therefore, the total saponins of eight Chinese herbal medicines commonly used to regulate abnormal glucose and lipid metabolism were selected as the research objects to observe their effect on the expression of HCBP6. Then, the three-dimensional structure of HCBP6 was predicted, followed by molecular docking with saponins in eight Chinese herbal medicines to quickly find potential active components. The results showed that all total saponins tended to up-regulate HCBP6 mRNA and protein expression, where gypenosides showed the optimum effect on up-regulating HCBP6 mRNA and ginsenosides showed the optimum effect on up-regulating HCBP6 protein expression. Reliable protein structures were obtained after the prediction of protein structures using the Robetta website and the evaluation of the predicted structures by SAVES. The saponins from the website and literature were also collected and docked with the predicted protein, and the saponin components were found to have good binding activity to the HCBP6 protein. The results of the study are expected to provide ideas and methods for the discovery of new drugs from Chinese herbal medicines to regulate glucose and lipid metabolism.
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Humains , Glucose , Métabolisme lipidique , Simulation de docking moléculaire , Médicaments issus de plantes chinoises/pharmacologie , Ginsénosides , Protéines , Saponines , ARN messagerRésumé
OBJECTIVE To establish the fingerprint of total saponins from Mussaenda pubescens, and to study the spectrum- effect relationship of its hepatoprotective activity. METHODS Ten batches of total saponins from M. pubescens from different origins were prepared using 75% ethanol as solvent. High-performance liquid chromatography (HPLC) and the Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints (2012 edition) were used to draw the fingerprints of 10 batches of total saponins from M. pubescens. The similarity evaluation and identification of common peaks were conducted. The same HPLC method was adopted to determine the contents of five triterpenoid saponins (mussaendoside H, mussaendoside U, mussaglaoside C, mussaendoside G and mussaendoside O). The hepatoprotective effect of total saponins from M. pubescens was investigated by establishing carbon tetrachloride-induced acute liver injury model mice, and the spectrum-effect relationship was studied by using grey correlation analysis. RESULTS There were 11 common peaks in 10 batches of total saponins from M. pubescens, 5 of which were identified, i.e. mussaendoside H (peak 3), mussaendoside U (peak 7), mussaglaoside C (peak 8), mussaendoside G (peak 9) and mussaendoside O (peak 11); the similarities of 10 batches of samples ranged 0.940- 0.991. Average contents of mussaendoside H, mussaendoside U, mussaglaoside C, mussaendoside G, mussaendoside O were 0.01- 0.05, 0.10-0.21, 0.10-0.18, 0.03-0.08, 0.20-0.40 mg/g, respectively. Ten batches of total saponins from M. pubescens could generally reduce the contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in liver tissue of model mice (P<0.05 or P<0.01). The E-mail:13878195336@139.com correlation between the common peak areas and the contents of ALT, AST, TNF-α, IL-6 and IL-1β were 0.602-0.757, 0.585-0.833, 0.593-0.795, 0.618-0.820, 0.607-0.804, respectively; the peaks with high correlation were peaks 11, 9 and 8 in order. CONCLUSIONS Ten batches of total saponins from M. pubescens have similar components, and the average contents of mussaendoside H, mussaendoside U, mussaglaoside C, mussaendoside G and mussaendoside O are different. The batches of samples have a certain degree of hepatoprotective effect; mussaendoside O, mussaendoside G and mussaglaoside C may be its main active components.
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Objective@#To explore the therapeutic effects of ginseng total saponins (GTSs) on cognitive impairments in astronauts caused by prolonged exposure to microgravity environment.@*Methods@#Fifty specific pathogen-free (SPF) male Wistar rats were randomized into control, hindlimb suspension (HLS), Huperzine A (HLS-Hup A 0.1 mg/kg), low-dose GTSs (HLS-GTSs 100 mg/kg), and high-dose GTSs (HLS-GTSs 200 mg/kg) groups, based on the completion time of reward-directed conditioning tasks. Except for rats in the control group, the others were subjected to HLS and treated with drugs (day 20 – 58), received reflex test under the condition of rewarding, and underwent Nissl body staining and Western blot detection on hippocampal.@*Results@#After modeling, rats in HLS group exhibited a reduction in the number of lever presses and an increase in the completion time of the reward-directed operant conditioning task Ⅰ (P < 0.05) when compared with the control group, which were not substantially altered in the HLS-GTSs 100 and 200 mg/kg groups (P > 0.05). In the reward-directed operant conditioning task Ⅱ, the HLS group rats demonstrated a marked decrease in the number of lever presses (P < 0.05) and nose pokes (P < 0.01) when compared with the control group rats; the HLS-GTSs 100 mg/kg showed a significant increase in the number of lever presses and nose pokes (P < 0.05), while the HLS-GTSs 200 mg/kg demonstrated a significant reduction in completion time and an elevation in the number of lever presses (P < 0.05) when compared with the HLS group rats. In visual signal discrimination task, compared with the control group rats, the HLS group rats showed decrease in the indexes of the visual signal discrimination(P < 0.01), while HLS-GTSs 100 and 200 mg/kg groups exhibited manifest increase in it (P < 0.01). In reward extinction experiment, the number of lever presses in HLS rats significantly increased when compared with the control group (P < 0.01); compared with the HLS group, HLS-GTSs 100 and 200 mg/kg groups demonstrated a marked descrease (P < 0.05). The expressions of N-methyl-D-aspartic acid receptor 1 (NR1) and phosophorylated N-methyl-Daspartic acid receptor 2B (p-NR2B) proteins were markedly decreased in rats in the HLS group (P < 0.05 and P < 0.01, respectively), while that of NR2B protein maintained the same (P > 0.05). GTSs increased the expression levels of p-NR2B (P < 0.01).@*Conclusion@#GTSs improved the learning and memory ability of complex operations by regulating the NR1/NR2B phosphorylation pathways in rats.
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Aim To investigate the effects of total saponins from Trillium tschonoskii maxim(TST)on cognitive impairment and mitochondrial autophagy in aging rats induced by D-galactose(D-gal). Methods Male SD rats were randomly divided into normal control group,model group(D-gal,subcutaneous injection),intervention group(TST,low,medium and high dose groups by intragastric administration),with 10 rats in each group,and administered for 6 weeks. Morris water maze was used to evaluate the cognitive function. HE and Nissl staining were used to test the hippocampal and brain cortex morphology. Immunohistochemistry staining was applied to detect the localization expression of Pink1 and Parkin. Western blot was employed to detect the expressions of Pink1,Parkin,LC3-Ⅱ,p62 and Beclin1. Results Compared with the normal control group,the escape latency time was prolonged and the number of crossing platform decreased in D-gal model group(P<0.05). The number of neurons in hippocampus significantly decreased. The positive cells labeled by Pink1 and Parkin staining in hippocampus significantly decreased. The expressions of Pink1,Parkin,LC3-Ⅱ and Beclin1 were markedly reduced,while the expression of p62 was significantly raised(P<0.05). Compared with D-gal model group,the escape latency time of TST dose groups was shortened,the Times of crossing the platform was more,and the time of staying in the platform quadrant increased(P<0.05). The number of neurons in hippocampus significantly increased. The positive cells labeled by Pink1 and Parkin staining in hippocampus significantly increased. The expressions of Pink1,Parkin,LC3-Ⅱ and Beclin1 in hippocampus were apparently up-regulated,while the protein expression of p62 was evidently down-regulated(P<0.05). Conclusions TST has neuroprotective effects on the learning and memory capacities in aging rats induced by D-gal,which may be related to the increasing levels of Pink1,Parkin,LC3-Ⅱ and Beclin1 proteins and the activation of mitochondrial autophagy.
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OBJECTIVE To study the osteoprotective effects and possible mechanism of total saponins of Chaenomeles speciosa on rheumatoid arthritis (RA) model mice, and to provide reference for further development of anti-RA drugs. METHODS Seventy male DBA/1 mice were randomly divided into normal group, model group, low-dose and high-dose groups of C. speciose total saponins (60, 240 mg/kg), Tripterygium wilfordii polyglycoside tablets group (positive control, 30 mg/kg), with 14 mice in each group. In addition to the normal group, the other groups of mice were induced by glucose-6-phosphate isomerase mixed polypeptide to prepare RA model. The body weight, rear toes thickness and arthritis scores of each group were recorded; the synovial inflammation, bone and cartilage destruction of ankle joint tissues were observed by hematoxylin-eosin staining, tartrate- resistant acid phosphatase staining and safranin O-fast green staining; the contents of interleukin-6 (IL-6) in serum and tumor necrosis factor α (TNF-α), IL-4 and IL-10 in ankle joint tissues were detected by ELISA; the expression levels of receptor activator of nuclear factor-κB ligand (RANKL), receptor activator of nuclear factor-κB (RANK), osteoprotegerin (OPG), tumor necrosis factor receptor-associated protein 6 (TRAF6) and nuclear factor of activated T cells 1 (NFATC1) protein in ankle joint tissues were detected by Western blot assay. RESULTS At the end of administration, compared with normal group, the body mass of mice in the model group was significantly reduced (P<0.05), and the arthritis score and the thickness of the left and right rear toes were significantly increased (P<0.05); the ankle joint tissues of mice in the model group showed significant synovial proliferation and inflammatory infiltration, the number of osteoclasts increased significantly and significant destruction of cartilage tissue. The content of IL-6 in serum, the content of TNF-α, the protein expression levels of RANKL, RANK, TRAF6 and NFATC1 in the ankle joint tissues were increased significantly (P<0.05), while the contents of IL- 4 and IL-10, the protein expression level of OPG in the ankle joint tissues were decreased significantly (P<0.05). Compared with model group, above pathomorphological changes and the content/level of indicators of mice in each administration group were significantly improved (P<0.05). CONCLUSIONS Total saponins of C. speciosa may exert osteoprotective effects on RA model mice, the mechanism of which may be associated with reducing the contents of IL-6 and TNF-α, increasing the contents of IL-4 and IL-10, inhibiting the activation of RANKL/RANK/OPG signal pathway, thus inhibiting the proliferation of osteoclasts and promoting the repair of cartilage and bone tissue.
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OBJECTIVE@#To investigate the protective effects of total saponins from Panax japonicus (TSPJ) against high-fat dietinduced testicular Sertoli cell junction damage in mice.@*METHODS@#Forty male C57BL/6J mice were randomized into normal diet group, high-fat diet group, and low-dose (25 mg/kg) and high-dose (75 mg/kg) TSPJ treatment groups (n=10). The mice in the normal diet group were fed a normal diet, while the mice in the other groups were fed a high-fat diet. After TSPJ treatment via intragastric administration for 5 months, the testes and epididymis of the mice were collected for measurement of weight, testicular and epididymal indices and sperm parameters. HE staining was used for histological evaluation of the testicular tissues and measurement of seminiferous tubule diameter and seminiferous epithelium height. The expression levels of ZO-1, occludin, claudin11, N-cadherin, E-cadherin and β-catenin in Sertoli cells were detected with Western blot, and the localization and expression levels of ZO-1 and β-catenin in the testicular tissues were detected with immunofluorescence assay. The protein expressions of LC3B, p-AKT and p-mTOR in testicular Sertoli cells were detected using double immunofluorescence assay.@*RESULTS@#Treatment with TSPJ significantly improved high-fat diet-induced testicular dysfunction by reducing body weight (P < 0.001), increasing testicular and epididymal indices (P < 0.05), and improving sperm concentration and sperm viability (P < 0.05). TSPJ ameliorated testicular pathologies and increased seminiferous epithelium height of the mice with high-fat diet feeding (P < 0.05) without affecting the seminiferous tubule diameter. TSPJ significantly increased the expression levels of ZO-1, occludin, N-cadherin, E-cadherin and β-catenin (P < 0.05) but did not affect claudin11 expression in the testicular tissues. Immunofluorescence assay showed that TSPJ significantly increased ZO-1 and β-catenin expression in the testicular tissues (P < 0.001), downregulated LC3B expression and upregulated p-AKT and p-mTOR expressions in testicular Sertoli cells.@*CONCLUSION@#TSPJ alleviates high-fat diet-induced damages of testicular Sertoli cell junctions and spermatogenesis possibly by activating the AKT/mTOR signaling pathway and inhibiting autophagy of testicular Sertoli cells.
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Mâle , Animaux , Souris , Souris de lignée C57BL , Testicule , Cellules de Sertoli , bêta-Caténine , Alimentation riche en graisse , Occludine , Protéines proto-oncogènes c-akt , Graines , Cadhérines , Jonctions intercellulairesRésumé
OBJECTIVE@#To reveal the neuroprotective effect and the underlying mechanisms of a mixture of the main components of Panax notoginseng saponins (TSPN) on cerebral ischemia-reperfusion injury and oxygen-glucose deprivation/reoxygenation (OGD/R) of cultured cortical neurons.@*METHODS@#The neuroprotective effect of TSPN was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, flow cytometry and live/dead cell assays. The morphology of dendrites was detected by immunofluorescence. Middle cerebral artery occlusion (MCAO) was developed in rats as a model of cerebral ischemia-reperfusion. The neuroprotective effect of TSPN was evaluated by neurological scoring, tail suspension test, 2,3,5-triphenyltetrazolium chloride (TTC) and Nissl stainings. Western blot analysis, immunohistochemistry and immunofluorescence were used to measure the changes in the Akt/mammalian target of rapamycin (mTOR) signaling pathway.@*RESULTS@#MTT showed that TSPN (50, 25 and 12.5 µ g/mL) protected cortical neurons after OGD/R treatment (P<0.01 or P<0.05). Flow cytometry and live/dead cell assays indicated that 25 µ g/mL TSPN decreased neuronal apoptosis (P<0.05), and immunofluorescence showed that 25 µ g/mL TSPN restored the dendritic morphology of damaged neurons (P<0.05). Moreover, 12.5 µ g/mL TSPN downregulated the expression of Beclin-1, Cleaved-caspase 3 and LC3B-II/LC3B-I, and upregulated the levels of phosphorylated (p)-Akt and p-mTOR (P<0.01 or P<0.05). In the MCAO model, 50 µ g/mL TSPN improved defective neurological behavior and reduced infarct volume (P<0.05). Moreover, the expression of Beclin-1 and LC3B in cerebral ischemic penumbra was downregulated after 50 µ g/mL TSPN treatment, whereas the p-mTOR level was upregulated (P<0.05 or P<0.01).@*CONCLUSION@#TSPN promoted neuronal survival and protected dendrite integrity after OGD/R and had a potential therapeutic effect by alleviating neurological deficits and reversing neuronal loss. TSPN promoted p-mTOR and inhibited Beclin-1 to alleviate ischemic damage, which may be the mechanism that underlies the neuroprotective activity of TSPN.
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Animaux , Rats , Bécline-1 , Encéphalopathie ischémique/métabolisme , Glucose , Infarctus du territoire de l'artère cérébrale moyenne/traitement médicamenteux , Mammifères/métabolisme , Neuroprotection , Neuroprotecteurs/usage thérapeutique , Oxygène , Panax notoginseng , Protéines proto-oncogènes c-akt/métabolisme , Lésion d'ischémie-reperfusion/métabolisme , Saponines/usage thérapeutique , Sérine-thréonine kinases TOR/métabolismeRésumé
OBJECTIVE@#To investigate the therapeutic effects of total saponins from Panax notognseng (PNS) combined with cyclophosphamide (CTX) in mice bearing hepatocellular carcinoma H22 cell xenograft.@*METHODS@#We examined the effects of treatment with different concentrations of PNS on H22 cell proliferation for 24 to 72 h in vitro using CCK8 colorimetric assay. Annexin V/PI double fluorescence staining was used to detect the effect of PNS on apoptosis of H22 cells. Mouse models bearing H22 cell xenograft were established and treated with CTX (25 mg/kg), PNS (120, 240 or 480 mg/kg), alone or in combinations. After treatments for consecutive 10 days, the mice were euthanized for examinations of carbon clearance ability of the monocytes and macrophages, splenic lymphocyte proliferation, tumor necrosis factor (TNF-α), interleukin-2 (IL-2), serum hemolysin antibody level, blood indicators, and the tumor inhibition rate.@*RESULTS@#Treatment with PNS concentration-dependently inhibited the proliferation and significantly promoted apoptosis of cultured H22 cells (P < 0.01). In the tumor-bearing mouse models, PNS alone and its combination with CTX both resulted in obvious enhancement of phagocytosis of the monocyte-macrophages, stimulated the proliferation of splenic lymphocytes, promoted the release of TNF-α and IL-2 and the production of serum hemolysin antibody, and increased the number of white blood cells, red blood cells and lymphocytes in the peripheral blood. Treatment with 480 mg/kg PNS combined with CTX resulted in a tumor inhibition rate of 83.28% (P < 0.01) and a life prolonging rate of 131.25% in the mouse models (P < 0.05).@*CONCLUSION@#PNS alone or in combination with CTX can improve the immunity and tumor inhibition rate and prolong the survival time of H22 tumor-bearing mice.
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Animaux , Humains , Souris , Carcinome hépatocellulaire/anatomopathologie , Cyclophosphamide/usage thérapeutique , Hémolysines , Hétérogreffes , Interleukine-2 , Tumeurs du foie/anatomopathologie , Panax notoginseng , Saponines/usage thérapeutique , Facteur de nécrose tumorale alphaRésumé
ObjectiveThe effect of inoculation with different organophosphate-resolving bacteria or compound bacteria on the quality of Paris polyphylla var. yunnanensis medicinal materials and rhizosphere soil fertility were studied to provide a reference for the development and application of biological bacterial fertilizer in artificial cultivation of P. polyphylla var. yunnanensis. MethodThe three dominant species of organophosphate-solubilizing bacteria were inoculated separately and in combination in sterilized soil by single-factor indoor pot planting, and no inoculation was used as the control group. The effect of inoculation of organophosphate-solubilizing bacteria on total saponins content in rhizomes of P. polyphylla var. yunnanensis, as well as microbial numbers, enzyme activities and nutrient contents in rhizosphere soil were analyzed. ResultIn the seven treatments inoculated with organophosphate-solubilizing bacteria, the total saponin content in the rhizomes of Paris polyphylla var. yunnanensis was increased by 16.42%, 3.83%, 16.86%, 33.69%, 2.11%, 13.44%, and 28.83%, respectively, compared with the control. Inoculation with organophosphate-solubilizing bacteria increased the number of soil microorganisms to varying degrees, and the effects of S6 and S7 treatments were the most significant. Inoculation with organophosphate-solubilizing bacteria improved the enzyme activity, and the effect of S7 treatment was the most significant. The activities of acid phosphatase, neutral phosphatase, alkaline phosphatase, protease, invertase and catalase were 49.96% and 104.67% , 110.17%, 99.61%, 26.26%, 11.29% higher than those of the control, respectively. Inoculation with organophosphate-solubilizing bacteria reduced the pH of the rhizosphere soil and increased the content of soil available nutrients. Under the S7 treatment, the contents of alkaline hydrolyzable nitrogen, available phosphorus and available potassium in the rhizosphere soil of P. polyphylla var. yunnanensis were 181.46%, 51.64% and 42.62% higher than those of the control, respectively. Correlation analysis showed that there was a significant positive correlation between total saponins and phosphatase activities, a significant positive correlation between soil microorganisms and soil enzyme activities, and a very significant positive correlation between soil nutrients. ConclusionInoculation of different organophosphate-resolving bacteria or compound bacteria can improve the quality and rhizosphere soil fertility of P. polyphylla var. yunnanensis. Among them, the mixed inoculation of three kinds of bacteria and the mixed inoculation of B. mycoides and B. wiedmannii have the best effect.
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Aim To discuss the effect of miR-199/ SIRT1/MFN2 signaling pathway on the progression of NASH and its related mechanisms.Methods 45 BALB/e mice were randomly divided into normal group, high fat diet(HFD) group, total saponins of panax japonicas ( TSPJ ) low-dose group ( 15 mg • kg-1) and TSPJ high-dose group (45 mg • kg"1 ).Normal group was given normal diet, while HFD group, TSPJ low-dose and high-dose groups were given high-fat diet.The mice were intragastrioally given 15 and 45 mg 'kg"1 TSPJ (dissolved in saline) daily in TSPJ low-dose and high-dose groups, while those in other groups were intragastric ally given the same a- mount of saline daily.After seven months, they were sacrificed for serum collection and hepatic tissue col¬lection.Results HE staining showed that liver lipido¬sis and inflammation were obvious in HFD group.while liver lipidosis anrl inflammation were alleviated in TSPJ group.MFN2 and SIRT1 levels significantly de¬creased.TNF-a, 1L-1 p , SREBP, ChREBP levels sig¬nificantly increased in HFD group.After treated with TSPJ, SIRT1 and MFN2 levels were significantly up- regulated , while TNF-a, IL-ip, ChREBP and SREBP levels were significantly down-regulated.The Immuno¬fluorescence results showed that the fluorescence inten¬sity of MFN2 and SIR 11 increased in TSPJ low-dose and high-dose groups.At mRNA level, miR-199 had a negative regulatory relationship with SIRT1.Conclu¬sions TSPJ can alleviate NASH induced by high fat diet through miR-199/SIRTl/MFN2 signaling path¬way.
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Non-alcoholic steatohepatitis(NASH) was induced by high-sugar and high-fat diet in mice to investigate the intervention effect of total saponins from Panax japonicus(TSPJ) and explore its possible mechanism. Mice were fed with high-sugar and high-fat diet to establish NASH model, and intervened with different doses of TSPJ(15, 45 mg·kg~(-1)). The animals were fed for 26 weeks. The histomorphology and pathological changes of liver tissues were observed by HE staining. The transcriptional expression levels of miR-199 a-5 p, autophagy related gene 5(ATG5) and inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α) in mouse liver were measured by quantitative Real-time polymerase chain reaction(qRT-PCR). Western blot was used to detect the expression of autophagy-related proteins ATG5, P62/SQSTM1(P62), and microtubule-associated protein light chain 3(LC3)-I/Ⅱ proteins in mouse liver. The expression of P62 protein was detected by immunofluorescence staining. In order to verify the targeting regulation relationship between miR-199 a-5 p and ATG5, miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor were transfected into Hepa 1-6 cells, and the expression of ATG5 mRNA and protein was detected. pMIR-reportor ATG5-3'UTR luciferase reporter gene plasmid was constructed and co-transfected with miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor into Hepa 1-6 cells to detect luciferase activity. In vivo, HE staining in the model group showed typical fatty degeneration and inflammatory infiltration, with increased expression of miR-199 a-5 p and decreased expression of ATG5 mRNA and protein. The expression of autophagy-associated protein P62 increased significantly, the ratio of LC3Ⅱ/Ⅰ decreased, and the transcriptional expression of inflammatory factors increased significantly. After the intervention by TSPJ, the pathological performance of liver tissue was significantly improved, the expression of miR-199 a-5 p decreased and the expression of ATG5 mRNA and protein increased, the expression of autophagy-associated protein P62 decreased significantly, the ratio of LC3Ⅱ/Ⅰ increased, and the transcriptional expression of inflammatory cytokines IL-6, IL-1β and TNF-α decreased significantly. In vitro, it was found that the expression of ATG5 mRNA and protein and luciferase activity decreased significantly in miR-199 a-5 p overexpression cells, while after inhibition of miR-199 a-5 p expression, the expression level of ATG5 mRNA and protein and luciferase activity increased. The results showed that TSPJ can improve NASH in mice fed with high-sugar and high-fat diet, and its mechanism may be related to the regulation of miR-199 a-5 p/ATG5 signal pathway, the regulation of autophagy activity and the improvement of inflammatory response of NASH.
Sujets)
Animaux , Souris , Autophagie , Protéine-5 associée à l'autophagie , microARN/génétique , Stéatose hépatique non alcoolique/génétique , Panax , Saponines/pharmacologieRésumé
OBJECTIVE:To stu dy the improvement effects o f Trillium tschonos kii total sapo nins(TTM)on adjuvant-induced arthritis model rats and its mechanism. METHODS :SD rats were randomly divided into model group (n=44)and blank group (n=6). Model group was given Complete Freund ’s adjuvant to induce the adjuvant-induced arthritis model ;blank group was given constant volume of normal saline with same method. Model group were divided into model group ,Tripterygium wilfordii polyglycoside tablets group (40 mg/kg),TTM low-dose ,medium-dose and high-dose groups (50,100,200 mg/kg),with 6 rats in each group. Administration groups were given relevant medicine intragastrically ;blank group and model group were given normal saline intragastrically ,once a day ,for consecutive 9 days. After last intragastric administration ,the body weight and foot swelling degree (bilateral)were detected ,and arthritis score was performed. The contents of tumor necrosis fator α(TNF-α), interleukin 1β(IL-1β)and IL- 6 were detected ;the pathomorphological changes of rat ankle were observed ;protein expression of NOD-like receptor protein 3 (NLRP3),apoptosis associated spot-like protein (ASC),caspase-1 were detected ;the protein expression of NLRP 3 in synovial tissue of ankle joint were also determined. RESULTS :Compared with blank group ,foot swelling degree,serum contents of TNF-α,IL-1β and IL-6,the protein expression of NLRP 3,ASC and caspase- 1 in knee tissue as well as the protein expression of NLRP 3 in synovial tissue of ankle joint were increased significantly in model group (P<0.05 or P< 0.01),while the body weight was decreased significantly (P<0.05). The obvious proliferation of synovial cells ,the congestion and inflammatory cell infiltration of synovial tissue were all observed. Compared with model group ,most of the above indexes were all reversed significantly in TTM groups (P<0.05 or P<0.01);the pathological changes such as synovial cell proliferation , congestion of synovial tissue and chondrocyte destruction were all relieved ,and inflammatory cell infiltration was alleviated.CONCLUSIONS:TTM can improve rheumatoid arthritis of rats, the mechanism of which may be associated with 用。E-mail:yulingling@ctgu.edu.cn inhibiting the activity of NLRP 3/caspase-1 signaling pathway and decreasing the expression of inflammatory factors TNF-α,IL-1β,IL-6.
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Objective:To illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis. Method:A total of 72 male SD rats were randomly divided into four groups: normal group, model group, total saponin group (160 mg·kg<sup>-1</sup>), celecoxib group (43.3 mg·kg<sup>-1</sup>), with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin(HE) staining. Immunohistochemistry method was used to detect the protein expression change of CD68, interleukin-4(IL-4), inducible nitric oxide synthase (iNOS) and transforming growth factor-<italic>β</italic><sub>1</sub>(TGF-<italic>β</italic><sub>1</sub>). Result:HE staining results showed that the inflammation of the model group was most obvious on the third day after modeling, and the disease was in the acute stage. On day 5, the inflammation was alleviated, and on day 8, the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue, and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd,5th and 8th day of administration (<italic>P</italic><0.01). Compared with the model group, the total saponins group could reduce the expression of CD68 and iNOS (<italic>P</italic><0.05,<italic>P</italic><0.01)on the 3rd day of administration, and significantly reduced them expression on the 5th and 8th days (<italic>P</italic><0.01). Compared with the normal group, IL-4 and TGF-<italic>β</italic><sub>1</sub> expression were increased in the model group when the drug was given for three days(<italic>P</italic><0.01). Total saponin group could enhance IL-4 expression(<italic>P</italic><0.05)and decreased the TGF-<italic>β</italic><sub>1</sub> expression(<italic>P</italic><0.01). Compared with normal group, the expression of IL-4 in the model group decreased on the 5th and 8th day of administration (<italic>P</italic><0.01), and the expression of TGF-<italic>β</italic><sub>1</sub> in the model group decreased on the 5th day of administration(<italic>P</italic><0.01). Compared with the model group, the total saponins group could increase the expression of IL-4 and TGF-<italic>β</italic><sub>1</sub> at 5 d and 8 d after administration (<italic>P</italic><0.01). Conclusion:Total saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.
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Objective:To investigate the effect of astragaloside Ⅳ(AST Ⅳ)and Notoginseng total saponins (NTS) combined with bone marrow mesenchymal stem cell (BMSC) transplantation on neural repair and angiogenesis in rats with cerebral ischemia. Method:The rats were randomly divided into a sham operation group, a model group, low- and high-dose AST Ⅳ + NTS groups, a BMSC infusion group, and low- and high-dose BMSC infusion+AST Ⅳ (10 and 20 mg·kg<sup>-1</sup>) + NTS group (25, 50 mg·kg<sup>-1</sup>). BMSCs were isolated and purified by whole bone marrow adherent culture. The positive expression of surface markers of BMSCs (CD29, CD90, CD34, and CD45) was detected by flow cytometry. The focal cerebral ischemia model was established by middle cerebral artery occlusion (MCAO). The PKH26-labeled BMSCs were injected into the tail vein of rats in the BMSC infusion group, once a day. The rats in the combination groups received BMSC injection once a day and intragastric administration of drugs twice a day. Other groups were administered twice a day by gavage. The sham operation group and the model group received the same amount of normal saline. Symptoms and signs of neurological deficits were assessed by the Longa method and the cerebral infarction rate was determined by TTC staining. The survival and vascularization [double positive expression of PKH26/vascular endothelial growth factor (VEGF)] after transplantation of BMSCs were observed by the immunofluorescence method. The protein expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was measured by Western blot. Result:BMSCs were properly isolated and cultured. The identification of surface markers CD29, CD90, CD34, and CD45 was consistent with the characteristics of BMSCs. The neurological deficit score and cerebral infarction rate of the model group were significantly increased (<italic>P</italic><0.01). All drugs and cell transplantation could alleviate the above pathological changes in varying degrees. The strongest effect was observed in high-dose BMSC infusion+AST Ⅳ+NTS group (<italic>P</italic><0.01), which was superior to those in the AST Ⅳ+NTS groups or the BMSC infusion group. BMSC injection helped cells survive in the ischemic brain tissues and promoted angiogenesis, and this effect could be enhanced by the combination with drugs. After cerebral ischemia, the expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was increased, and the effect in the BMSC infusion+AST Ⅳ+NTS groups was the strongest (<italic>P</italic><0.01). Conclusion:AST Ⅳ combined with NTS can promote the survival of transplanted BMSCs and facilitate angiogenesis after target repair of damaged blood vessels after cerebral ischemia. The mechanism may be related to the improvement of the local microenvironment in the brain after cerebral ischemia and the promotion of the survival and differentiation of transplanted stem cells.
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Objective:To study the effects of different arbuscular mycorrhizal(AM) fungi combinations on the growth and quality of<italic> Paris polyphylla</italic> var. <italic>yunnanensis</italic> seedlings,in order to provide reference for the cultivation of high quality <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic>. Method:Through the room temperature pot inoculation test method,nine groups of different AM fungi combinations were inoculated into sterilized soil,and the control group was not inoculated. The effects of different AM fungi combinations on root infection rate,photosynthetic parameters,physiological indexes and chemical components of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> seedlings were observed. Result:The spore density in rhizosphere soil,the infection rate and intensity of AM fungi in roots,and the activities of succinate dehydrogenase and alkaline phosphatase in roots of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> were significantly increased by inoculating the combination of exogenous AM fungi. Compared with the control group,the contents of chlorophyll,malondialdehyde,soluble sugar,soluble protein content and the activities of antioxidant enzymes in leaves of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> were increased in different AM fungi treatment groups. The content of total saponin increased from T1 to T2,decreased rapidly from T2 to T4,and increased at T5. Conclusion:Different mixed AM fungus can form a good symbiotic relationship with the roots of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic>,improve the mycorrhizal vitality,enhance the resistance of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> to adverse environment,promote the growth and development of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic> and improve the quality of rhizome. According to the comprehensive inoculation effect,the combination of S5,S8 and S9 AM fungus has the best effect,which provides reference value for the application of mycorrhizal biotechnology of <italic>P. polyphylla</italic> var. <italic>yunnanensis</italic>.