Evidence-based Complementary and Alternative Medicine
·
2018
·
IF 2.064
Renjie Xu,Yichu Yuan,Jia Qi,Jia Zhou,Xiaowen Guo,Jian Zhang,Ruanjuan Zhan
Loganin, iridoid glycosides, is the main bioactive ingredients in the plant Strychnos nux-vomica L. and demonstrates various pharmacological effects, though poor oral bioavailability in rats. In this study, the intestinal absorption mechanism of loganin was investigated using the human intestinal Caco-2 cell monolayer model in both the apical-to-basolateral (A-B) and the basolateral-to-apical (B-A) direction; additionally, transport characteristics were systematically investigated at different concentrations, pHs, temperatures, and potential transporters. The absorption permeability (AB) of loganin, which ranged from 12.17 to 14.78 × 10−6cm/s, was high at four tested concentrations (5, 20, 40, and 80μM), while the major permeation mechanism of loganin was found to be passive diffusion with active efflux mediated by multidrug resistance-associated protein (MRP) and breast cancer resistance protein (BCRP). In addition, it was found that loganin was not the substrate of efflux transporter P-glycoprotein (P-gp) since the selective inhibitor (verapamil) of the efflux transporter exhibited little effects on the transport of loganin in the human intestinal Caco-2 cells. Meanwhile, transport from the apical to the basolateral side increased 2.09-fold after addition of a MRP inhibitor and 2.32-fold after addition of a BCRP inhibitor. In summary, our results clearly demonstrate, for the first time, a good permeability of loganin in the human intestinal Caco-2 cell model and elucidate, in detail, the intestinal absorption mechanism and the effects of transporters on iridoid glycosides compounds.
研究领域:
药理学
·
生物医学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD SCIENCE
·
2024
·
IF 3.2
Qian Du,Ruiyan Wang,Zeyuan Deng,Jianqun Zhou,Nan Li,Wenwen Li,Liufeng Zheng
Sucrose emerges as a chelating agent to form a stable sucrose-metal-ion chelate that can potentially improve metal-ion absorption. This study aimed to analyze the structure of sucrose-calcium chelate and its potential to promote calcium absorption in both Caco-2 monolayer cells and mice. The characterization results showed that calcium ions mainly chelated with hydroxyl groups in sucrose to produce sucrose-calcium chelate, altering the crystal structure of sucrose (forming polymer particles) and improving its thermal stability. Sucrose-calcium chelate dose dependently increased the amount of calcium uptake, retention, and transport in the Caco-2 monolayer cell model. Compared to CaCl 2 , there was a significant improvement in the proportion of absorbed calcium utilized for transport but not retention (93.13 ± 1.75% vs. 67.67 ± 7.55%). Further treatment of calcium channel inhibitors demonstrated the active transport of sucrose-calcium chelate through Cav1.3. Cellular thermal shift assay and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays indicated that the ability of sucrose-calcium chelate to promote calcium transport was attributed to its superior ability to bind with PMCA1b, a calcium transporter located on the basement membrane, and stimulate its gene expression compared to CaCl 2 . Pharmacokinetic analysis of mice confirmed the calcium absorption-promoting effect of sucrose-calcium chelate, as evident by the higher serum calcium level (44.12 ± 1.90 mg/L vs. 37.42 ± 1.88 mmol/L) and intestinal PMCA1b gene expression than CaCl 2 . These findings offer a new understanding of how sucrose-calcium chelate enhances intestinal calcium absorption and could be used as an ingredient in functional foods to treat calcium deficiency. Practical Application The development of high-quality calcium supplements is crucial for addressing the various adverse symptoms associated with calcium deficiency. This study aimed to prepare a sucrose-calcium chelate and analyze its structure, as well as its potential to enhance calcium absorption in Caco-2 monolayer cells and mice. The results demonstrated that the sucrose-calcium chelate effectively promoted calcium absorption. Notably, its ability to enhance calcium transport was linked to its strong binding with PMCA1b, a calcium transporter located on the basement membrane, and its capacity to stimulate PMCA1b gene expression. These findings contribute to a deeper understanding of how the sucrose-calcium chelate enhances intestinal calcium absorption and suggest its potential use as an ingredient in functional foods for treating calcium deficiency.
研究领域:
营养学
·
药理学
·
细胞生物学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MEDIATORS OF INFLAMMATION
·
2021
·
IF 4.711
Huan Lan,Lu-Ying Zhang,Wen He,Wan-Ying Li,Zhen Zeng,Bo Qian,Chengqiang Wang,Jia-Le Song
The integrity and permeability of the intestinal epithelial barrier are important indicators of intestinal health. Impaired intestinal epithelial barrier function and increased intestinal permeability are closely linked to the onset and progression of various intestinal diseases. Sinapic acid (SA) is a phenolic acid that has anti-inflammatory, antihyperglycemic, and antioxidant activities; meanwhile, it is also effective in the protection of inflammatory bowel disease (IBD), but the specific mechanisms remain unclear. Here, we evaluated the anti-inflammatory of SA and investigated its potential therapeutic activity in LPS-induced intestinal epithelial barrier and tight junction (TJ) protein dysfunction. SA improved cell viability; attenuated epithelial permeability; restored the protein and mRNA expression of claudin-1, ZO-1, and occludin; and reversed the redistribution of the ZO-1 and claudin-1 proteins in LPS-treated Caco-2 cells. Moreover, SA reduced the inflammatory response by downregulating the activation of the TLR4/NF-κB pathway and attenuated LPS-induced intestinal barrier dysfunction by decreasing the activation of the MLCK/MLC pathway. This study demonstrated that SA has strong anti-inflammatory activity and can alleviate the occurrence of high intercellular permeability in Caco-2 cells exposed to LPS.
研究领域:
药理学
·
免疫学
·
细胞生物学
·
胃肠病学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2022
·
IF 8.025
Weibo Zhang,Shucheng Zhang,Chong Chen,Ning Liu,Dong Yang,Pengjie Wang,Fazheng Ren
Pea albumin (PA) can reach the intestine in the active form because it is highly resistant to gastric acid and proteolytic enzymes after their oral intake, which can supply various bioactivities. However, there is no detailed knowledge of the intestinal cell uptake about PA. The aim of this work was to study the internalization mechanism and intracellular trafficking route of PA. The uptake of PA-cyanine 5.5 NHS ester (Cy5.5) was a time-dependent and concentration-dependent process in Caco-2 cells. Endocytosis inhibitors or small interfering RNA (siRNA) techniques revealed that the internalization of PA-Cy5.5 was energy-dependent and mediated by caveolin-mediated endocytosis. Furthermore, we observed colocalization of PA-Cy5.5 and its subcellular localization in Caco-2 cells by using confocal laser scanning microscopy, which revealed that the intracellular trafficking process of PA-Cy5.5 was related to endoplasmic reticulum, Golgi, and lysosome. Interestingly, PA can alleviate lipopolysaccharide -induced ER stress, which may be the main reason why pea albumin is anti-inflammatory. Overall, our findings suggest caveolin may be critical for PA uptake in enterocytes and could contribute to explore the bioactivities mechanism of pea albumin in body.
研究领域:
营养学
·
细胞生物学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2022
·
IF 8.025
Yun-yun Shao,Yi-nan Zhao,Yi-fan Sun,Yao Guo,Xiao Zhang,Zhuang-peng Chang,Rui-gang Hou,Jianping Gao
For Codonopsis Radix polysaccharides (CRPs), oral administration is generally considered the most convenient route for patients. However, the details of its absorption and transport mechanisms remain unclear. In this study, we aimed to evaluate the oral absorption of CPA (an inulin-type fructan extracted from CRPs) in mice and Caco-2 cells. It was labeled with fluorescein isothiocyanate, and the fluorescence derivative (FCPA) was used to trace the behavior of CPA. The results showed that FCPA could be absorbed after oral administration and has a wide tissue distribution, including in the stomach, intestine, kidneys, and liver. FCPA was poorly absorbed, and its internalization was time- and energy-dependent, as well as dependent on cholesterol- and dynamin-mediated endocytosis. Confocal laser scanning microscopy showed successful cellular internalization of FCPA from the cytoplasm to the nucleus. In addition, we found that FCPA was trafficked to endosomes and lysosomes, and that tubulin was required for its intracellular transport. These findings add new details to our knowledge of the internalization and transport mechanisms of CPA, which may prove useful to the development and application of oral formulations of CRPs.
研究领域:
药理学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2021
·
IF 7.963
Wantang Huang,Hua Yin,Yuanyu Yang,Lizhu Jin,Guining Lu,Zhi Dang
Microplastics (MPs) pollution becomes an emergent threat to the ecosystem, and its joint effect with organic contaminants will cause more severe consequences. Recently, MPs has been observed in human feces, suggesting that we are exposed to an uncertain danger. In this study, the joint effect of polyethylene microplastics particles (PEMPs) and Tetrabromobisphenol A (TBBPA) on human gut was explored through the simulation experiment in vitro with human cell Caco-2 and gut microbiota. The toxicity of TBBPA and PEMPs on Caco-2 human cells was considered by physiological and biochemical indexes such as cell proliferation, cell cycle, reactive oxygen species, lactate dehydrogenase release, and mitochondrial membrane potential. Besides, microbial community diversity, community structure, and function changes of gut microbiota were investigated using Illumina 16S rRNA gene MiSeq sequencing to reveal the influence of TBBPA and PEMPs on human gut microbiota. The results indicated that both PEMPs and TBBPA would deteriorate the status of Caco-2 cells, and TBBPA played a major role in it; meanwhile, PEMPs affected Caco-2 cells at high concentrations. Particularly, TBBPA and PEMPs exhibited a joint effect on Caco-2 cells to a certain degree. TBBPA selectivity inhibited the growth of gram-positive bacteria such as Enterococcus and Lactobacillus , contributing to the thriving of gram-negative bacteria such as Escherichia and Bacteroides . The existence of PEMPs would enhance the proportion of Clostridium , Bacteroides , and Escherichia . Community composition changed dramatically with the interference of PEMPs and TBBPA; this was undesirable to the healthy homeostasis of the human gut. PICRUSt analysis determined both PEMPs and TBBPA interfered with the metabolism pathways of gut microbiota. Hence, the threat of MPs and TBBPA to humans should arouse vigilance.
研究领域:
环境科学
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毒理学
·
微生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Zixing Chen,Chi Ho Jacky Chau,Andrew Erickson,Huafang Ding,Shouhe Huang,Wen-Sen He,Hanyue Zhu,Zhen-Yu Chen
Phytosterols have long been used to prevent hypercholesterolemia-related diseases, although their effects on intestinal cholesterol metabolism remain elusive. This study applied comparative proteomics to investigate the effects of various sterols on proteomic changes in differentiated Caco-2 cells. A total of 206, 231, 180, and 197 differentially expressed proteins were identified following treatment with cholesterol, β-sitosterol, stigmasterol, and campesterol, respectively. Functional enrichment analysis revealed that phytosterols significantly altered the proteome of Caco-2 cells, though their effect on cholesterol transport protein expression was negligible. Notably, while phytosterols did not regulate the expression of acyl-CoA:cholesterol acyltransferase 2 (ACAT2), β-sitosterol and stigmasterol significantly lowered the level of cholesterol oleate in Caco-2 cells by 57.1 ± 0.8% and 51.4 ± 3.9%, respectively, demonstrating that they inhibited ACAT2-catalyzed cholesteryl esterification activity. In view of the fact that cholesterol needs to be converted to cholesteryl esters before incorporation into chylomicrons, it was concluded that phytosterols primarily reduce intestinal cholesterol absorption by inhibiting ACAT2 enzyme activity rather than its gene expression. This study provides new insights into the regulation of cholesterol homeostasis by phytosterols in the small intestine.
研究领域:
营养生物化学
·
分子生物学
·
细胞生物学
·
脂质代谢
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药理学
·
蛋白质组学
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产品关联
DOI
PubMed
高影响因子
Journal of Pharmaceutical Analysis
·
2020
·
IF 2.673
Siqi Ren,Jingjing Liu,Yunwen Xue,Mei Zhang,Qiwei Liu,Jie Xu,Zunjian Zhang,Rui Song
Saikosaponins (SSs) are the main active components extracted from Bupleuri Radix (BR) which has been used as an important herbal drug in Asian countries for thousands of years. It has been reported that the intestinal bacteria plays an important role in the in vivo disposal of oral SSs. Although the deglycosylated derivatives (saikogenins, SGs) of SSs metabolized by the intestinal bacteria are speculated to be the main components absorbed into the blood after oral administration of SSs, no studies have been reported on the characteristics of SGs for their intestinal absorption, and those for SSs are also limited. Therefore, a rapid UHPLC-MS/MS method was developed to investigate and compare the apparent permeability of three common SSs (SSa, SSd, SSb 2 ) and their corresponding SGs (SGF, SGG, SGD) through a bidirectional transport experiment on Caco-2 cell monolayer model. The method was validated according to the latest FDA guidelines and applied to quantify the six analytes in transport medium samples extracted via liquid-liquid extraction (LLE). The apparent permeability coefficient ( P app ) determined in this study indicated that the permeability of SGs improved to the moderate class compared to the corresponding parent compounds, predicting a higher in vivo absorption. Moreover, the efflux ratio (ER) value demonstrated an active uptake of SSd and the three SGs, while a passive diffusion of SSa and SSb 2 .
研究领域:
药理学
·
药剂学
·
分析化学
·
中药学
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产品关联
DOI
PubMed
高影响因子
PLANTA MEDICA
·
2021
·Vol. 87
·期 07
·570-580
·
IF 3.356
Wei Shen,Xiaolong Hu,Yajun Niu,Yimeng Lu,Baolin Wang,Hao Wang
Abrus mollis is commonly used as a traditional Chinese medicine for the treatment of liver diseases due to its hepatoprotection and anti-inflammation, but the absorption properties of its main bioactive ingredients remain unclear. Our previous studies verified that the flavonoid C-glycosides, including vicenin-2 (1), isoschaftoside (2), and schaftoside (3), were the major active components in A. mollis for hepatic protection against nonalcoholic fatty liver disease, hepatitis, and hepatic fibrosis. This study investigated the bioaccessibility and transport mechanisms of total flavonoid C-glycoside, as well as vicenin-2 (1), isoschaftoside (2), and schaftoside (3), in A. mollis by simulated digestion and use of the Caco-2 cell model. Moreover, this study attempted to verify their absorption properties by in situ gastrointestinal perfusion in rats. Total flavonoid C-glycoside and 1, 2, and 3 exhibited similar bioaccessibility of 84.58%, 85.13%, 83.05%, and 81.65% respectively after simulated digestion. The transport of total flavonoid C-glycoside in the Caco-2 cell model increased with the concentration, and the transport showed saturation characteristics with the time and concentration of total flavonoid C-glycoside to a certain degree. The Papp values of total flavonoid C-glycoside and the 3 flavonoid C-glycosides were significantly improved by verapamil, probenecid, and EDTA-Na2. Their absorption properties in the gastrointestinal tract were consistent with that found in Caco-2 cells, and superior absorption rates were observed in the duodenum and jejunum. The absorption pattern of total flavonoid C-glycoside may involve multiple transport pathways, including active transport, passive diffusion, and the paracellular pathway. TFC was actively pumped out by P-glycoprotein and multidrug resistance-associated protein. These results revealed that the bioaccessibility and intestinal absorption characteristic of total flavonoid C-glycoside were consistent with the 3 major flavonoids.
研究领域:
药理学
·
中药学
·
消化与吸收机制
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMICAL & PHARMACEUTICAL BULLETIN
·
2021
·Vol. 69
·期 11
·1054-1060
·
IF 1.645
Yuqin Zhou,Weitong Hu,Xiangli Zhang,Yi Wang,Wenya Zhuang,Fengzhi Li,Qingyong Li
In the evaluation of the druggability of candidate compounds, it was vital to predict the oral bioavailability of compounds from apparent permeability (Papp) across Caco-2 cell-culture model of intestinal epithelium cultured on commercial transwell plate inserts. The study was to investigate the transport characteristics and permeability of FL118 (10, 11-Methylenedioxy-20(S)-camptothecin) derivatives 7-Q6 (7-(4-Ethylphenyl)-10, 11-methylenedioxy-20(S)-camptothecin) and 7-Q20 (7-(4-Trifluoromethylphenyl)-10, 11-methylenedioxy-20(S)-camptothecin). Transport characteristics and permeability of the tested compounds to the small intestine were assessed at different concentrations (0.5, 1 µM) via Caco-2 cell monolayers model in vitro. Uptake studies based on Caco-2 cells, including temperatures, concentrations, and the influence of efflux transporters, were combined to confirm the transport characteristics of the tested compounds. Furthermore, cytotoxicity results showed that the concentrations used in the experiments were non-toxic and harmless to cells. In addition, The Papp of 7-Q6 was (3.69 ± 1.07) × 10−6 cm/s with efflux ratio (ER) 0.98, while the Papp of 7-Q20 was (7.78 ± 0.89) × 10−6 cm/s with ER 1.05 for apical-to-basolateral (AP→BL) at 0.5 µM, suggesting that 7-Q20 might possess higher oral bioavailability in vivo. Furthermore, P-glycoprotein (P-gp) was proved to slightly affect the accumulations of 7-Q20, while the absorption of 7-Q6 was irrelevant with P-gp and breast cancer resistant protein (BCRP) based on the cellular uptake assays. Accordingly, 7-Q6 was completely absorbed by passive diffusion, and 7-Q20 was mainly dependent on passive diffusion with being effluxed by P-gp slightly. Meanwhile, both 7-Q6 and 7-Q20 were potential antitumor drugs that might exhibit high oral bioavailability in the body.
研究领域:
药理学
·
药物递送
·
细胞生物学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
International Journal of Peptide Research and Therapeutics
·
2018
·Vol. 25
·期 4
·1319-1331
·
IF 1.132
Liu Huifan,Wu Hui,Wang Qin
In a previous study a crude peptide extract was obtained from Dendrobium aphyllum , and this study focused on the functional characterization of the peptide fraction (named DPP) after being purified by DEAE Sepharose. The DPP with high purity contained 16 types of free amino acids and 6 polypeptides. DPP could act as the AAPH-free radical scavenger to protect the erythrocyte under oxidative stress, and the attenuation significantly increased after intestinal digestion. Moreover, the results of the Caco-2 cell models indicated the high absorptive and transfer rates of DPP, in which the passive diffusion was the major transfer pathway. Through the LC-TOF/MS-MS analysis, the peptide sequences Asp-Asp-Asp-Tyr and Asp-Tyr-Asp-Asp were very sensitive to the gastric environment and were completely degraded by Caco-2 cells. While Ser-Ser-Arg, Ala-Val-Thr-Phe , and Met-Leu-Cys-Ala resisted the gastric condition; and sequence Met-Leu-Cys-Ala maintained its structure after the subsequent intestinal digestion and the Caco-2 cells’ uptake.
研究领域:
生物化学
·
营养学
·
药理学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2018
·
IF 6.434
Kaiyu Jiang,Caihuan Huang,Rui Jiao,Weibin Bai,Jie Zheng,Shiyi Ou
Acrolein (ACR), glyoxal (GO), methylglyoxal (MGO), hydroxymethylfurfural (HMF), and malondialdehyde (MDA) are toxic contaminants for humans. This work aimed to investigate whether intake of proteins can mitigate their toxicity. Simulated gastrointestinal digestion of proteins from pork, chicken, milk powder and soy protein isolate eliminated amount of ACR, GO, MGO, HMF, and MDA. Among six amino acids, cysteine showed highest capacity for elimination of these toxic compounds through the formation of adducts; it reached the highest elimination capacity for GO, MGO, ACR, MDA, and HMF in 40 min at pH 2.0, and 20 min at pH 7.0. The formed adducts between cysteine and GO, MGO, or ACR showed much lower toxicity against Caco-2 cells. Incubation of the cells with 8 mM GO and MGO for 48 h decreased the cell viability to 16.1%, 16.9% respectively; while incubation of the same concentration of their adducts still kept the cell viability at 82.2% and 81.6% respectively. Cysteine showed much higher detoxifying capacity for ACR than GO and MGO, which can lower the toxicity of ACR toward Caco-2 cells by 80 times.
研究领域:
食品科学
·
毒理学
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）