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This study aimed to investigate the biological effects and underlying mechanisms of the total ginsenosides from Panax ginseng stems and leaves on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in mice. Sixty male C57BL/6J mice were randomly divided into a control group, a model group, the total ginsenosides from P. ginseng stems and leaves normal administration group(61.65 mg·kg~(-1)), and low-, medium-, and high-dose total ginsenosides from P. ginseng stems and leaves groups(15.412 5, 30.825, and 61.65 mg·kg~(-1)). Mice were administered for seven continuous days before modeling. Twenty-four hours after modeling, mice were sacrificed to obtain lung tissues and calculate lung wet/dry ratio. The number of inflammatory cells in bronchoalveolar lavage fluid(BALF) was detected. The levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in BALF were detected. The mRNA expression levels of IL-1β, IL-6, and TNF-α, and the levels of myeloperoxidase(MPO), glutathione peroxidase(GSH-Px), superoxide dismutase(SOD), and malondialdehyde(MDA) in lung tissues were determined. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in lung tissues. The gut microbiota was detected by 16S rRNA sequencing, and gas chromatography-mass spectrometry(GC-MS) was applied to detect the content of short-chain fatty acids(SCFAs) in se-rum. The results showed that the total ginsenosides from P. ginseng stems and leaves could reduce lung index, lung wet/dry ratio, and lung damage in LPS-induced ALI mice, decrease the number of inflammatory cells and levels of inflammatory factors in BALF, inhibit the mRNA expression levels of inflammatory factors and levels of MPO and MDA in lung tissues, and potentiate the activity of GSH-Px and SOD in lung tissues. Furthermore, they could also reverse the gut microbiota disorder, restore the diversity of gut microbiota, increase the relative abundance of Lachnospiraceae and Muribaculaceae, decrease the relative abundance of Prevotellaceae, and enhance the content of SCFAs(acetic acid, propionic acid, and butyric acid) in serum. This study suggested that the total ginsenosides from P. ginseng stems and leaves could improve lung edema, inflammatory response, and oxidative stress in ALI mice by regulating gut microbiota and SCFAs metabolism.
Subject(s)
Mice , Male , Animals , Ginsenosides/pharmacology , Tumor Necrosis Factor-alpha/metabolism , Interleukin-6 , Panax/genetics , Lipopolysaccharides/adverse effects , Gastrointestinal Microbiome , RNA, Ribosomal, 16S , Mice, Inbred C57BL , Acute Lung Injury/genetics , Lung/metabolism , Superoxide Dismutase/metabolism , Plant Leaves/metabolism , RNA, MessengerABSTRACT
A content determination method based on ~1H-qNMR was developed for the determination of total ginsenosides in Shenmai Injection. The parameters were optimized with CD_3OD as the solvent, dimethyl terephthalate as the internal standard, the peak at δ 8.11 as the internal standard peak, and the peaks at δ 1.68 and δ 0.79 as quantitative peaks of total ginsenosides. The developed ~1H-qNMR-based method was validated methodologically. The results showed that the method could achieve accurate measurement of total ginsenosides in Shenmai Injection in the range of 0.167 6-3.091 1 mmol·L~(-1). The developed ~1H-qNMR-based method for total ginsenosides is simple in operation, short in analysis time, strong in specificity, independent of accompanying standard curve, and small in sample volume, which can serve as a reliable mean for the quality control of Shenmai Injection. This study is expected to provide new ideas for the development of quantification methods of total ginsenosides.
Subject(s)
Drug Combinations , Drugs, Chinese Herbal , Ginsenosides/analysis , Quality ControlABSTRACT
OBJECTIVE:To study the improvement effects of total ginsenosides on the senescence of PC 12 cells induced by D-galactose and its mechanism. METHODS :Rat pheochromocytoma (PC12)cells were treated with D-galactose to establish cell senescence model. CCK- 8 method was used to screen the D-galactose modeling concentration and total ginsenosides concentration. Normal control group ,model group ,total ginsenosides low and high concentration groups were set up. Cell senescence ,cell apoptosis rate ,apoptotic cycle and mitochondrial membrane potential (MMP),cell adenosine triphosphate (ATP)and reactive oxygen species (ROS)levels in each group were detected. The expression of apoptosis related proteins [B lymphoma 2(Bcl-2)and its related egg X protein (Bax),cytochrome C (Cyt-C)] and oxidative damage related proteins [nuclear factor 2 related factor 2 (Nrf2),heme oxygenase 1(HO-1)] were detected. In addition ,positive drug group [ 5 mmol/L N-acetyl-L-cysteine(NAC)] and positive control group [ D-galactose+5 mmol/L NAC] were set up to compare the levels of oxidative damage related proteins. RESULTS:D-galactose could significantly inhibit the survival rate of PC 12 cells,with a critical concentration of 20 mg/mL. The total ginsenosides could significantly increase the survival rate of D-galactose induced senescent cells with a median effective concentration(EC50)of 65 μg/mL,and then the low and high concentrations of total ginsenosides were set at 55 and 65 μg/mL. Compared with normal control group ,the number of aging cells increased ,the apoptotic rate and percentage of G 1 phase were significantly increased i n model group. the percentage of S phase ,MMP and ATP contents ,the protein expression of Bcl- 2 and Cyt-C in mitochondria were decreased significantly ,whileROS content ,the protein expression of Bax ,Nrf2 and Cyt-C protein in endochylema were increased significantly (P<0.05 or P<0.01). Compared with model group ,the number of E-mail:sunqiao150509@163.com aging cells reduced ,the apoptosis rates and percentage of G 1 phase were significantly decreased in total ginsenosides low and high concentration groups ,the percentage of S phase ,the contents of MMP and ATP (except for low concentration group ),protein expression of Bcl- 2,Nrf2 and HO- 1 as well as protein expression of Cyt-C in mitochondria were increased significantly ;ROS level (except for low concentration group )and Bax protein as well as protein expression of Cyt-C were decreased significantly. The protein expression of Nrf 2 and HO- 1 were increased significantly in positive control group (P<0.05 or P<0.01), but it was lower than that of total ginsenosides groups . CONCLUSIONS:Total ginsenosides can improve D-galactose induced senescence of P 12 cells,the mechanism of which may be related to activating Nrf 2 antioxidant signal pathway to antagonize D-galactose induced oxidative stress and alleviating mitochondrial dysfunction.
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Objective To investigate the therapeutic effect of total saponins of ginseng (GFS)combined with serotonin re-uptake inhibitor(SSRIs)fluoxetine on rats with myocardial infarction(MI)and depression.Methods Left thoracotomy and per-manent ligation of left atrial appendage were performed to establish MI model in SD rats ,and depression model was established by chronic uncontrollable mild stress.SD rats were randomly divided into control group ,MI depression group(MI+ D group) (intragastric perfusion with 2 mL saline) ,fluoxetine group(F group)(intragastric perfusion with 0.2% fluoxetine at 10 mL/kg) and ginseng fruit saponins+ fluoxetine group(GFS+ F group)(intragastric perfusion of 20 mL/kg ginsenoside + 10 mL/kg 0.2% fluoxetine). The depression degree ,survival rate ,cardiac function ,electrophysiological parameters and histological chan-ges were evaluated after 28 days of intervention.Results Compared with the control group ,the consumption of sugar water in MI+D group was significantly decreased ,and the frequency of climbing ,crawling distance and crawling speed were significantly lower than those of the control group.GFS+fluoxetine and fluoxetine significantly increased the consumption of sugar water , increased the frequency of climbing and crawling distance ,but the treatment of GFS+fluoxetine was better than fluoxetine ,and could also improve the crawl speed. There was no significant difference in survival rate between MI +D group ,F group and GFS+F group(P=0.24).Compared with the control group ,LVEDD ,LVESD and LVWT were significantly increased ,LVEF and FS significantly reduced in the MI+D group.GFS+fluoxetine and fluoxetine significantly reduced LVEDD in MI+D group and increased LVEF ,but GFS+ fluoxetine was superior to fluoxetine ,and also decreased LVESD and LVWT.Compared with the control group ,VERP and APD90 in MI+D group were significantly prolonged and VFT was significantly decreased.GFS+flu-oxetine and fluoxetine significantly improved the VFT of MI+D rats ,and the effect of GFS+fluoxetine was better than that of fluoxetine ,and they could shorten VERP and APD90 in model group. There was no significant difference in myocardial infarct size between MI+ D group ,F group and GFS+ F group(P= 0.076).Fluoxetine intervention did not improve the myocardial thickness and collagen content in the myocardial infarction area ,but GFS+ fluoxetine did.Conclusion Fluoxetine and GFS+fluoxetine can improve depression ,cardiac function and electrophysiological parameters of the rats with MI and depression ,but the effect of GFS+fluoxetine is better than that of fluoxetine ,and the myocardial thickness and collagen expression also have been improved by GFS+fluoxetine.
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Objective:To study the biotransformation rule of the chemical components of total ginsenosides from ginseng stems and leaves tranformed by submerged fermentation of ganoderma lucidum,and to lay the foundation for further development and research of the medicinal fungus of ginseng stems and leaves.Methods:The total ginsenosides from ginseng stems and leaves were fermented by ganoderma lucidum,the changes of chemical components before and after fermentation were measured by thin-layer chromatography;the levels of total ginsenosides from ginseng stems and leaves before and after fermentation were detected by ultraviolet spectrophotometry;the levels of ginsenoside Rb1,Rg3,Rd,CK,Re,Rg,and Rh1 before and after fermentation were measured by high performance liquid chromatography.The human hepatocellular carcinoma SMMC-7721 cells were divided into blank control group,low,medium and high doses of total ginsenosides from ginseng stems and leaves groups and medicinal fungus of ginseng stems and leaves groups.The dosages of total ginsenosides from ginseng stems and leaves and medicinal fungus of ginseng stems and leaves were 5,10 and 20 mg·L-1,respectively.The inhibitory rate(IR) of the cells was determined by MTT assay.Results:The results of thin-layer chromatography showed that the saponins before and after fermentation occured mutual transformation;the levels of total ginsenosides from ginseng stems and leaves before and after fermentation were 749.98 and 602.26 mg·g-1,respectively;the levels of panaxadiol saponins Rb1,Rg3,Rd and CK before fermentation were 24.54,2.21,87.22 and 0 mg·g-1,and the levels of panaxatriol saponins Re,Rg and Rh1 were 151.34,77.02,and 3.06 mg·g-1.After fermentation,the level of Rb1 was decreased by 61.45%,the levels of Rg3,Rd and CK were increased by 238.91%,34.43% and 268.00%.The levels of Re and Rg1 were decreased by 63.75% and 64.41%,and the level of Rh1 was increased by 408.88%;the IR of the SMMC-7721 cells in medium dose of medicinal fungus of ginseng stems and leaves group was higher than that in medium dose of total ginsenosides from ginseng stems and leaves group(P<0.05);the IR of the SMMC-7721 cells in high dose of medicinal fungus of ginseng stems and leaves group was higher than that in high dose of total ginsenosides from ginseng stems and leaves group(P<0.01).Conclusion:The total ginsenosides from ginseng stems and leaves transformed by the submerged fermentation of ganoderma lucidum can significantly change its chemical components,which have stronger anti-proliferation activity in the SMMC-7721 cells.
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Objective: To study the chemotaxis response of Fusarium solani and Cylindrocarpon destructans on total ginsenosides. Methods: Three chemotactic parameters (concentration, temperature, and pH) were determined by plate assay and spore germination method to research the chemotaxis response of two pathogens and their spores. Results: It showed that F. solani had strong chemotactic response at the low concentration of total ginsenosides, and the data of chemotactic mobile index (CMI) was 1.2948, SGR was 66%, chemotaxis growth rate (CGR) was 0.533, and (mycelial growth) MG was 0.5220 mg/mL. However, C. destructans had strong chemotactic response at the middle concentration of total ginsenosides, and the data of CMI was 1.2556, SGR was 63%, CGR was 0.465, and MG was 0.449 4 mg/mL. Conclusion: The low and middle concentration (0.2-20 mg/L) of total ginsenosides had the significant promoting effect on chemotaxis response of F. solani, and the chemotaxis response of C. destructans happened at different concentration of ginsenosides, and the SGR, MGR, and the amount of MG of these two pathogens had also been significantly improved, whereas the chemotaxis response effect decreases as the ginsenosides concentration increases.
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OBJECTIVE To observe the protective effect and underlying mechanism of total ginsenosides (TG) on bleomycin-induced pulmonary fibrosis. METHODS Intratracheal instillation of bleomycin 5 mg · kg-1 was conducted to establish a pulmonary fibrosis mouse model. Kunming mice(1/2 males and 1/2 females)were randomly divided into sham-operation(Sham),model,total ginsen?osides 40,80 and 160 mg·kg-1 and prednisone acetate(5 mg·kg-1) groups. After 28 d administration,the histopathological changes in the lung were analyzed by hematoxylin eosin(HE)and Masson staining. The exprssion of alpha smooth muscle actin(α-SMA)in the lung was detected by real-time PCR. The content of hydroxyproline(HYP)and glutathione(GSH),level of total antioxidant capacity(T-AOC)and hydroxy radical(·OH),activity of myeloperoxidase(MPO)and nitric oxide synthase(NOS)in the lung were detected by corresponding kits. RESULTS Compared with the sham group,the pulmonary indexes in model group were significantly increased(P<0.01),alveolitis and pulmonary fibrosis were obvious. The mRNA expression ofα-SMA,content of HYP and · OH,activity of MPO and NOS were increased(P<0.05),but the content of GSH and T-AOC in model group was decreased(P<0.05). Compared with model group,the pulmonary indexes in TG 80 and 160 mg · kg-1 and prednisone acetate 5 mg · kg-1 groups were reduced(P<0.05),and the degree of alveolitis and pulmonary fibrosis was mitigated. The mRNA expression ofα-SMA,content of HYP and · OH, the activity of MPO and NO were decreased (P<0.05),while the content of GSH and T-AOC was increased(P<0.05). CONCLUSION TG can improve the degree of mice pulmonary fibrosis induced by bleomycin. The mechanism may be related to the increased antioxidant capacity of organisms.
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Aim To investigate the effects of total gin-senosides ( TG) on microvascular regeneration and car-diac function in rat after acute myocardial infarction ( AMI ) . Methods The acute myocardial infarction model was created with left coronary artery ligated in male Sprague Dawley rats. The model rats were ran-domly divided into sham, model, TG low and high dose groups. TG groups were injected into abdominal cavity with TG(20 mg·kg-1·d-1, 40 mg·kg-1· d-1 ) . Sham and model groups were injected with e-qual-volume normal saline. On the 35th day post-opera-tion, heart function was examined by color doppler ul-trasoundination. HE, Masson and immunohistochemis-trical staining were used to observe the histopathologi-cal changes of myocardium and micro vessel density. The level of vascular endothelial growth factor( VEGF) and basic fibroblast growth factor( bFGF) mRNA were detected by real-time PCR. Results Compared with the model group, high and low dose TG obviously de-creased the left ventricular end diastolic dimension, the left ventricular end-systolic dimension, the left ventric-ular end-diastolic volume and the left ventricular end-systolic volume(P<0. 05 and P<0. 01), and signifi-cantly increased the ejection fraction and the fractional shortening ( P <0. 01 ) . The histopathological changes of myocardium on myocardial infarction and fibrosis were dramatically reduced by TG. But ventricular wall was thicker. Two dose TG remarkably increased the expressions of VEGF and bFGF mRNA and micro ves-sel density of compositive CD31 + cells in ischemic myocardial tissue and around of infarct area(P<0. 01, P <0 . 0 5 ) . Conclusions TG could improve the car-diac function of acute myocardial infarction rat. The mechanism may be related to the upregulation of VEGF and bFGF gene expression, the promotion angiogene-sis, then the improvement of blood supply in myocardi-al infarction area.
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Objective: To investigate the contents of total ginsenosides and 10 sorts of monomer ginsenosides in the roots of Panax ginseng from different habitats of northeast China and its processed products, and thus to provide the useful reference data for its quality st andard establishment and st andardized cultivation. Methods: Based on the pharmacopoeia and literatures related to the roots of P. ginseng, the contents of total ginsenosides and 10 sorts of monomer ginsenosides in the roots of P. ginseng samples collected from 10 different habitats of northeast China and its processed product samples were studied and determined, and then these various indicators were analyzed by DTOPSIS method. Results: Ginseng from Changbai, Ji'an, and Fusong reached the st andard of Chinese Pharmacopoeia 2010 and Chinese National St andards in those different measurement indicators. Comprehensive evaluation of ginseng from Changbai, Ji'an, Fusong, and Jingyu by DTOPSIS method showed better results than others. Conclusion: The qualities of ginseng from different habitats of northeast China are different. Ginseng collected from those four habitats which result better quality of ginseng than that of ginseng from other place stems from Chinese GAP ginseng planting bases, and thus reflects the importance of GAP for ginseng cultivation.