Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (6): 1118-1126.DOI: 10.19852/j.cnki.jtcm.2024.06.003
• Research Articles • Previous Articles Next Articles
Renshen (Radix Ginseng) polysaccharide promotes repair of the mice intestinal mucosa through regulatory mechanisms based on polyamine and human antigen R
WANG Guanyu, DAI Xingzhen, LIU Yiting, ZHU Zeming, HU Ling, LI Ruliu(
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- Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, China
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Received:2024-02-22Accepted:2024-08-20Online:2024-12-15Published:2024-11-12 -
Contact:Prof. LI Ruliu, Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, China. lrl@gzucm.edu.cn Telephone: +86-20-36585444 -
Supported by:National Natural Science Foundation of China: Study on the Effect of Supplementing Qi and Invigorating the Spleen on the Migration of Small Intestinal Epithelial Cells through the Regulation of Polyamines and Calcium Ions(81673940);First-class Discipline Construction Major Project of Guangzhou University of Chinese Medicine, Guangzhou University of Chinese Medicine Planning (2020) No. 62: based on the Viscous Characteristics of Dampness and Pathogenic Factors in Lingnan, this Paper Studied the Differential Characteristics of the Syndrome of Deficiency of Spleen and Stomach Disease and the Intervention Mechanism of Lingnan Traditional Chinese Medicine for Mucosal Damage Repair
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WANG Guanyu, DAI Xingzhen, LIU Yiting, ZHU Zeming, HU Ling, LI Ruliu. Renshen (Radix Ginseng) polysaccharide promotes repair of the mice intestinal mucosa through regulatory mechanisms based on polyamine and human antigen R[J]. Journal of Traditional Chinese Medicine, 2024, 44(6): 1118-1126.
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Figure 1 Effect of Renshen (Radix Ginseng) polysaccharides on cell migration and the expression of HuR A: effect of Renshen (Radix Ginseng) polysaccharide on cell migration in the absence of load; B: effect of Renshen (Radix Ginseng) polysaccharides on cell migration under DFMO loading; C: effect of Renshen (Radix Ginseng) polysaccharides on cell migration under CMLD-2 loading. A: unloaded culture: blank control, positive drug control (PuT, 20 μmol/L), PGP (40, 80, 160 mg/L). B: DFMO loaded culture: blank control, DFMO model (DFMO at 5 mmol/L), positive drug control (PuT, 20 μmol/L + DFMO), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). C: CMLD-2 loaded culture: blank control, CMLD-2 model (CMLD-2 at 3 μmol/L), positive drug control (PuT, 20 μmol/L + CMLD-2), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). DFMO: difluoromethylornithine; PuT: putrescine; CMLD-2: 5,7-Dimethoxy-8-[1-(4-methoxyphenyl)-3-oxo-3-(1-pyrrolidinyl) propyl]-4-phenyl-2H-chromen-2-one; PGP: panax ginseng polysaccharide. All data was measured by one-way analysis, and Dunnett’s T3 test was performed for inter-group comparisons. The data measured in this research were expressed as mean ± standard deviation (n = 3). Compared to control group, aP < 0.01, bP < 0.05; compared to the model group, cP < 0.05.
Figure 1 Effect of Renshen (Radix Ginseng) polysaccharides on cell migration and the expression of HuR A: effect of Renshen (Radix Ginseng) polysaccharide on cell migration in the absence of load; B: effect of Renshen (Radix Ginseng) polysaccharides on cell migration under DFMO loading; C: effect of Renshen (Radix Ginseng) polysaccharides on cell migration under CMLD-2 loading. A: unloaded culture: blank control, positive drug control (PuT, 20 μmol/L), PGP (40, 80, 160 mg/L). B: DFMO loaded culture: blank control, DFMO model (DFMO at 5 mmol/L), positive drug control (PuT, 20 μmol/L + DFMO), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). C: CMLD-2 loaded culture: blank control, CMLD-2 model (CMLD-2 at 3 μmol/L), positive drug control (PuT, 20 μmol/L + CMLD-2), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). DFMO: difluoromethylornithine; PuT: putrescine; CMLD-2: 5,7-Dimethoxy-8-[1-(4-methoxyphenyl)-3-oxo-3-(1-pyrrolidinyl) propyl]-4-phenyl-2H-chromen-2-one; PGP: panax ginseng polysaccharide. All data was measured by one-way analysis, and Dunnett’s T3 test was performed for inter-group comparisons. The data measured in this research were expressed as mean ± standard deviation (n = 3). Compared to control group, aP < 0.01, bP < 0.05; compared to the model group, cP < 0.05.
Figure 2 Effect of Renshen (Radix Ginseng) polysaccharides on the distribution of HuR A1-A6: immunofluorescence assay (× 400): Alexa Fluor ?. HuR dyeing. B1-B6: DAPI dyeing. A1, B1: Control; A2, B2: DFMO of 5 mmol/L; A3, B3: DFMO of 5 mmol/L + PUT 20 μmol/L; A4, B4: DFMO of 5 mmol/L+PGP 40 mg/L; A5, B5: DFMO of 5 mmol/L + PGP 80 mg/L; A6, B6: DFMO of 5 mmol/L + PGP160 mg/L. Dyeing method of all pictures are the immunohistochemical DAB method. HuR: human antigen R; DFMO: difluoromethylornithine; PuT: putrescine; PGP: panax ginseng polysaccharide; CMLD-2: 5,7-Dimethoxy-8-[1-(4-methoxyphenyl)-3-oxo-3-(1-pyrrolidinyl) propyl]-4-phenyl-2H-chromen-2-one.
Figure 2 Effect of Renshen (Radix Ginseng) polysaccharides on the distribution of HuR A1-A6: immunofluorescence assay (× 400): Alexa Fluor ?. HuR dyeing. B1-B6: DAPI dyeing. A1, B1: Control; A2, B2: DFMO of 5 mmol/L; A3, B3: DFMO of 5 mmol/L + PUT 20 μmol/L; A4, B4: DFMO of 5 mmol/L+PGP 40 mg/L; A5, B5: DFMO of 5 mmol/L + PGP 80 mg/L; A6, B6: DFMO of 5 mmol/L + PGP160 mg/L. Dyeing method of all pictures are the immunohistochemical DAB method. HuR: human antigen R; DFMO: difluoromethylornithine; PuT: putrescine; PGP: panax ginseng polysaccharide; CMLD-2: 5,7-Dimethoxy-8-[1-(4-methoxyphenyl)-3-oxo-3-(1-pyrrolidinyl) propyl]-4-phenyl-2H-chromen-2-one.
Figure 3 Modulation of STIM1 by Renshen (Radix Ginseng) polysaccharides A: effect of Renshen (Radix Ginseng) polysaccharide on protein (A1) and mRNA (A2) expression of STIM1 in the absence of load; B: effect of Renshen (Radix Ginseng) polysaccharide on protein (B1) and mRNA expression (B2) of STIM1 during DFMO loading; C: effect of Renshen (Radix Ginseng) polysaccharides on post-transcriptional events of STIM1mRNA. A: unloaded culture: blank control, positive drug control (PuT, 20 μmol/L), PGP (40, 80, 160 mg/L). B, C: DFMO loaded culture: blank control, DFMO model (DFMO at 5 mmol/L), positive drug control (PuT, 20 μmol/L + DFMO), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). A1, B1: Weston blot was used to detect the expression of STIM1 protein. A2, B2: RT-qPCR was used to detect the expression of STIM1 mRNA. C1: blocking gene transcription with actinomycin D and detecting mRNA expression at different time points to assess STIM1 mRNA half-life. C2: double luciferase reporter gene system to detect the translation efficiency of STIM1mRNA. STIM1: stromal interaction molecule 1; DFMO: difluoromethylornithine; PuT: putrescine; PGP: panax ginseng polysaccharide; RT-qPCR: real-time reverse transcription-quantitative polymerase chain reaction. All data was measured by one-way analysis, and Dunnett’s T3 test was performed for inter-group comparisons. The data measured in this research were expressed as mean ± standard deviation (n = 3). Compared to control group, aP < 0.05, bP < 0.01; compared to the model group, cP < 0.05.
Figure 3 Modulation of STIM1 by Renshen (Radix Ginseng) polysaccharides A: effect of Renshen (Radix Ginseng) polysaccharide on protein (A1) and mRNA (A2) expression of STIM1 in the absence of load; B: effect of Renshen (Radix Ginseng) polysaccharide on protein (B1) and mRNA expression (B2) of STIM1 during DFMO loading; C: effect of Renshen (Radix Ginseng) polysaccharides on post-transcriptional events of STIM1mRNA. A: unloaded culture: blank control, positive drug control (PuT, 20 μmol/L), PGP (40, 80, 160 mg/L). B, C: DFMO loaded culture: blank control, DFMO model (DFMO at 5 mmol/L), positive drug control (PuT, 20 μmol/L + DFMO), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). A1, B1: Weston blot was used to detect the expression of STIM1 protein. A2, B2: RT-qPCR was used to detect the expression of STIM1 mRNA. C1: blocking gene transcription with actinomycin D and detecting mRNA expression at different time points to assess STIM1 mRNA half-life. C2: double luciferase reporter gene system to detect the translation efficiency of STIM1mRNA. STIM1: stromal interaction molecule 1; DFMO: difluoromethylornithine; PuT: putrescine; PGP: panax ginseng polysaccharide; RT-qPCR: real-time reverse transcription-quantitative polymerase chain reaction. All data was measured by one-way analysis, and Dunnett’s T3 test was performed for inter-group comparisons. The data measured in this research were expressed as mean ± standard deviation (n = 3). Compared to control group, aP < 0.05, bP < 0.01; compared to the model group, cP < 0.05.
Figure 4 Modulation of Cdc42 by Renshen (Radix Ginseng) polysaccharides A: effect of Renshen (Radix Ginseng) polysaccharide on protein (A1) and mRNA (A2) expression of Cdc42 in the absence of load; B: effect of Renshen (Radix Ginseng) polysaccharide on protein (B1) and mRNA expression (B2) of Cdc42 during DFMO loading; C: effect of Renshen (Radix Ginseng) polysaccharides on post-transcriptional events of Cdc42 mRNA. A: unloaded culture: blank control, positive drug control (PuT, 20 μmol/L), PGP (40, 80, 160 mg/L). B, C: DFMO loaded culture: blank control, DFMO model (DFMO at 5 mmol/L), positive drug control (PuT, 20 μmol/L + DFMO), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). A1, B1: Weston blot was used to detect the expression of Cdc42 protein. A2, B2: RT-qPCR was used to detect the expression of Cdc42 mRNA. C: double luciferase reporter gene system to detect the translation efficiency of Cdc42 mRNA. Cdc42: cell division cycle 42; DFMO: difluoromethylornithine; PuT: putrescine; PGP: panax ginseng polysaccharide; RT-qPCR: real-time reverse transcription-quantitative polymerase chain reaction. All data was measured by one-way analysis, and Dunnett’s T3 test was performed for inter-group comparisons. The data measured in this research were expressed as mean ± standard deviation (n = 3). Compared to control group, aP < 0.01, bP < 0.05; compared to the model group, cP < 0.05.
Figure 4 Modulation of Cdc42 by Renshen (Radix Ginseng) polysaccharides A: effect of Renshen (Radix Ginseng) polysaccharide on protein (A1) and mRNA (A2) expression of Cdc42 in the absence of load; B: effect of Renshen (Radix Ginseng) polysaccharide on protein (B1) and mRNA expression (B2) of Cdc42 during DFMO loading; C: effect of Renshen (Radix Ginseng) polysaccharides on post-transcriptional events of Cdc42 mRNA. A: unloaded culture: blank control, positive drug control (PuT, 20 μmol/L), PGP (40, 80, 160 mg/L). B, C: DFMO loaded culture: blank control, DFMO model (DFMO at 5 mmol/L), positive drug control (PuT, 20 μmol/L + DFMO), PGP (PGP, 40 mg/L + DFMO, 80 mg/L + DFMO, 160 mg/L + DFMO). A1, B1: Weston blot was used to detect the expression of Cdc42 protein. A2, B2: RT-qPCR was used to detect the expression of Cdc42 mRNA. C: double luciferase reporter gene system to detect the translation efficiency of Cdc42 mRNA. Cdc42: cell division cycle 42; DFMO: difluoromethylornithine; PuT: putrescine; PGP: panax ginseng polysaccharide; RT-qPCR: real-time reverse transcription-quantitative polymerase chain reaction. All data was measured by one-way analysis, and Dunnett’s T3 test was performed for inter-group comparisons. The data measured in this research were expressed as mean ± standard deviation (n = 3). Compared to control group, aP < 0.01, bP < 0.05; compared to the model group, cP < 0.05.
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