Background: The roots of Panax ginseng contain two types of tetracyclic triterpenoid saponins, namely, protopanaxadiol (PPD)-type saponins and protopanaxatiol (PPT)-type saponins. In P. ginseng, the protopanaxadiol 6-hydroxylase (PPT synthase) enzyme catalyses protopanaxatriol (PPT) production from protopanaxadiol (PPD). In this study, we constructed homozygous mutant lines of ginseng by CRISPR/Cas9-mediated mutagenesis of the PPT synthase gene and obtained the mutant ginseng root lines having complete depletion of the PPT-type ginsenosides. Methods: Two sgRNAs (single guide RNAs) were designed for target mutations in the exon sequences of the two PPT synthase genes (both PPTa and PPTg sequences) with the CRISPR/Cas9 system. Transgenic ginseng roots were generated through Agrobacterium-mediated transformation. The mutant lines were screened by ginsenoside analysis and DNA sequencing. Result: Ginsenoside analysis revealed the complete depletion of PPT-type ginsenosides in three putative mutant lines (Cr4, Cr7, and Cr14). The reduction of PPT-type ginsenosides in mutant lines led to increased accumulation of PPD-type ginsenosides. The gene editing in the selected mutant lines was confirmed by targeted deep sequencing. Conclusion: We have established the genome editing protocol by CRISPR/Cas9 system in P. ginseng and demonstrated the mutated roots producing only PPD-type ginsenosides by depleting PPT-type ginsenosides. Because the pharmacological activity of PPD-group ginsenosides is significantly different from that of PPT-group ginsenosides, the new type of ginseng mutant producing only PPD-group ginsenosides may have new pharmacological characteristics compared to wild-type ginseng. This is the first report to generate target-induced mutations for the modification of saponin biosynthesis in Panax species using CRISPR-Cas9 system.
In order to obtain the basic information for the development of ginseng varieties with high saponin contents. saponin contents and ginsenosides of Panax ginseng (Korean ginseng) and Panax quinquefolium (American ginseng) grown under the same environmental conditions were analysed. Crude saponin contents of root and aerial parts were more in Panax quinquefolium than in Panax ginseng, and aerial parts had more saponin contents in comparison with a root. Protopanaxatriol saponin was greatly more in the aerial parts of ginseng while more amount of protopanaxadiol saponins were detected in the root. As for the ginsenosides, the patterns of ginsenosides detected in total saponin of the aerial parts were not different between two species, Panax ginseng and Panax quinquefolium, but the root ginsenoside patterns were quite different. Ginsenosides such as Rg$_2$, R$_{f}$. R$_{a}$ and R$_{o}$ were not detected in the root of Panax quinquefolium (American ginseng).).).).
Background: Although many studies have evaluated the efficacy and pharmacokinetics of Korean Red Ginseng (KRG) components (Rg1, Rb1, Rg3, Rd, etc.), few have examined the in vivo pharmacokinetics of the radiolabeled components. This study investigated the pharmacokinetics of ginsenosides and their metabolite compound K (CK), 20(s)-protopanaxadiol (PPD), and 20(s)-protopanaxatriol (PPT) using radioisotopes in rat oral administration. Methods: Sprague-Dawley rats were dosed orally once with 10 mg/kg of the tritium(3H) radiolabeled samples, and then the blood was collected from the tail vein after 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 12, 24, 48, 96, and 168 h. Radioactivity in the organs, feces, urine, and carcass was determined using a liquid scintillation counter (LSC) and a bio-imaging analyzer system (BAS). Results and conclusion: After oral administration, as the 3H-labeled ginsenosides were converted to metabolites, Cmax and half-life increased, and Tmax decreased. Interestingly, Rb1 and CK showed similar values, and after a single oral administration of components, the cumulative excretion ratio of urine and feces was 88.9%-92.4%. Although most KRG components were excreted within 96-168 h of administration, small amounts of components were detected in almost all tissues and mainly distributed to the liver except for the digestive tract when observed through autoradiography. This study demonstrated that KRG components were distributed to various organs in the rats. Further studies could be conducted to prove the bioavailability and transmission of KRG components to confirm the mechanism of KRG efficacy.
Viral infections are known as one of the major factors causing death. Ginseng is a medicinal plant that demonstrated a wide range of antiviral potential, and saponins are the major bioactive ingredients in the genus Panax with vast therapeutic potential. Studies focusing on the antiviral activity of the genus Panax plant-derived agents (extracts and saponins) and their mechanisms were identified and summarized, including contributions mainly from January 2016 until January 2022. P. ginseng, P. notoginseng, and P. quinquefolius were included in the review as valuable medicinal herbs against infections with 14 types of viruses. Reports from 9 extracts and 12 bioactive saponins were included, with 6 types of protopanaxadiol (PPD) ginsenosides and 6 types of protopanaxatriol (PPT) ginsenosides. The mechanisms mainly involved the inhibition of viral attachment and replication, the modulation of immune response by regulating signaling pathways, including the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway, cystathionine γ-lyase (CSE)/hydrogen sulfide (H2S) pathway, phosphoinositide-dependent kinase-1 (PDK1)/ protein kinase B (Akt) signaling pathway, c-Jun N-terminal kinase (JNK)/activator protein-1 (AP-1) pathway, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway. This review includes detailed information about the mentioned antiviral effects of the genus Panax extracts and saponins in vitro and in vivo, and in human clinical trials, which provides a scientific basis for ginseng as an adjunctive therapeutic drug or nutraceutical.
Young Woo Kim;Seon Been Bak;Won-Yung Lee;Su Jin Bae;Eun Hye Lee;Ju-Hye Yang;Kwang Youn Kim;Chang Hyun Song;Sang Chan Kim;Un-Jung Yun;Kwang Il Park
Journal of Ginseng Research
/
v.47
no.3
/
pp.479-491
/
2023
Background: Hepatocellular carcinoma (HCC) has a high incidence and is one of the highest mortality cancers when advanced stage is proceeded. However, Anti-cancer drugs available for treatment are limited and new anti-cancer drugs and new ways to treat them are minimal. We examined that the effects and possibility of Red Ginseng (RG, Panax ginseng Meyer) as new anti-cancer drug on HCC by combining network pharmacology and molecular biology. Materials and Methods: Network pharmacological analysis was employed to investigate the systems-level mechanism of RG focusing on HCC. Cytotoxicity of RG was determined by MTT analysis, which were also stained by annexin V/PI staining for apoptosis and acridine orange for autophagy. For the analyze mechanism of RG, we extracted protein and subjected to immunoblotting for apoptosis or autophagy related proteins. Results: We constructed compound-target network of RG and identified potential pathways related to HCC. RG inhibited growth of HCC through acceleration of cytotoxicity and reduction of wound healing ability of HCC. RG also increased apoptosis and autophagy through AMPK induction. In addition, its ingredients, 20S-PPD (protopanaxadiol) and 20S-PPT (protopanaxatriol), also induced AMPK mediated apoptosis and autophagy. Conclusion: RG effectively inhibited growth of HCC cells inducing apoptosis and autophagy via ATG/AMPK in HCC cells. Overall, our study suggests possibility as new anti-cancer drug on HCC by proof for the mechanism of the anti-cancer action of RG.
Panax ginseng leaf tea was developed for the functional benefit of health, preference and convenience. The leaves of 4-year-old ginseng were selected in July and August. The ginseng leaf was treated by three methods : heat processed tea(HPT), aged tea(AGT) and hot-air dried tea(DRT). The contents and compositions of their crude saponin of ginseng leaves were measured. 1. The content of crude saponin of HPT was the higher than other treatments. The content of HPT was 18.72∼18.82%, ACT 18.24∼18.29% and DRT 17.02∼17.17%. 2. The harvest time and treatment methods were not affect the composition of ginsenoside in ginseng leaf tea. The ginsenoside-Re was shown the highest value as 1.97∼2.15. And ginsenoside-Rd was 1.48∼1.79, -Rg$_1$ 1.33∼1.58 and -Rb, -Rb$_2$, -Rc in the order. 3. The content of protopanaxadiol(PD) and protopanaxatriol(PT) was shown that DRT was 1.11∼1.13, HPT 1.09~l.12 and AGT 0.92∼1.02. The content of PD and PT were shown similar result at any harvest time. 4. The contents of crude saponin extracted by hot-water at 5 min was the higher ratios in HPT and harvested in July than other treatments. The content of crude saponin of ginseng leaf harvested in July was 15.88% and HPT was 16.88%. The order of contents of ginsenoside were -Re, -Rd, -Rg$_1$, -Rb$_1$, -Rb$_2$, and - Rc. The extraction ratio of crude saponin extracted by the circulated extraction method in 8 hours and 5 min extraction were 81.74∼84.38%. And HPT of ginseng leaf harvested in July was the highest value 84.3% but the extraction ratio of ginsenoside was 78.00~88.13%. But the extraction ratio of ginsenoside was similar trend in all treatments.
It has generally been accepted that quality of ginseng should be determined not by the content of a single component but by composition and balance of total active principles. However, there still can be an exception with a product in which a given ginsenoside is used for the treatment of a specific disease. Although ginsenosides have been regarded to be major active components of ginseng and employed as index components for the quality control, it does not consistent with the traditional concept on ginseng quality creterion; main root has been more highly appreciated than the lateral or fine root. Content of ginsenosides in the lateral or fine root is much higher than that in main root. However, the ratio of protopanaxadiol (PD) and protopanaxatriol (PT) saponins existing in various part of ginseng root is greatly different. The ratio of PD/PT saponins in main root is well balanced but the thinner the root is the higher the ratio. Thus far, a total of 34 different kinds of ginsenosides have been isolated from Korean (red) ginseng, and their pharmacological activities were elucidated partly. Interestingly, different ginsenoside shows similar or contrary effects to each other in biological systems, thus indicating the significance of absolute content of single ginsenoside as well as compositional patterns of each ginsenoside. Therefore, pharmacological activities of ginseng should be determined as a wholly concept. In these regards, standardization of ginseng material (fresh ginseng root) should be preceded to the standardization of ginseng products because ginsenoside content and non-saponin active principles such as polysaccharides and nitrogen (N)-containing compound including proteins are significantly different from part to part of the root. In other words, the main root contains less ginsenosides than other lateral or fine roots. Contents of polysaccharides and N-containing compound in main root is higher. However, the quality control of ginseng products focused on non-saponin compounds has limitation in applying to the analytical method, because of the difficult chemical analysis of these compounds. Content of ginsenosides, and ratios of PD/PT and ginsenoside Rb,/Rg, are inversely proportional to the diameter of ginseng root. Therefore, these can be served as the chemical parameters for the indirect method of evaluating from what part of the root does the material originate. Furthermore, contents of polysaccharides and N-containing compounds show inverse relationship to saponin content. Therefore, it seems that index for analytical chemistry of saponin can be applied to the indirect method of evaluating not only saponin but also non-saponin compounds of ginseng. From these viewpoints, it is strongly recommended that quality of ginseng or ginseng products be judged not only by the absolute content of given ginsenoside but also by varieties and compositional balance of ginsenosides, including contents of non-saponin active principles.
Channels formed by the co-assembly of the KCNQ1 subunit and the mink (KCNE1) subunit underline the slowly activating delayed rectifier $K^+$ channels ($I_{Ks}$) in the heart. This $K^+$ channel is one of the main pharmacological targets for the development of drugs against cardiovascular disease. Panax ginseng has been shown to exhibit beneficial cardiovascular effects. In a previous study, we showed that ginsenoside Rg3 activates human KCNQ1 $K^+$ channel currents through interactions with the K318 and V319 residues. However, little is known about the effects of ginsenoside metabolites on KCNQ1 $K^+$ alone or the KCNQ1 + KCNE1 $K^+$ ($I_{Ks}$) channels. In the present study, we examined the effect of protopanaxatriol (PPT) and compound K (CK) on KCNQ1 $K^+$ and $I_{Ks}$ channel activity expressed in Xenopus oocytes. PPT more strongly inhibited the $I_{Ks}$ channel currents than the currents of KCNQ1 $K^+$ alone in concentration- and voltage-dependent manners. The $IC_{50}$ values on $I_{Ks}$ and KCNQ1 alone currents for PPT were $5.18{\pm}0.13$ and $10.04{\pm}0.17{\mu}M$, respectively. PPT caused a leftward shift in the activation curve of $I_{Ks}$ channel activity, but minimally affected KCNQ1 alone. CK exhibited slight inhibition on $I_{Ks}$ and KCNQ1 alone $K^+$ channel currents. These results indicate that ginsenoside metabolites show limited effects on $I_{Ks}$ channel activity, depending on the structure of the ginsenoside metabolites.
In this paper, through the mutual interpretation and verification of the ancient Korean history books with different origin that have been suspected as false documents, it proves that they could be logically real records and reveal that the substance of the legendary 'medicine for anti-aging and longevity', which also had been mentioned in Chinese old books, is Korean ginseng. Furthermore, with reference to the modern Y chromosomal map of the migratory routes of mankind corresponding to these routes recorded in 「Budoji」, the core history book, the formation of the four ethnic constitution groups (Sasang Constitution) based on the life style of each human group has been estimated. And the cause of Korean ginseng with fever problem for Southeast Asians is their pharmacogenomic constitution problem by protopanaxatriol (PPT) type ginsenosides in ginseng. It was resolved with over production of protopanaxdiol (PPD) type ginsenosides against PPT type in Korean red ginseng as historical or scientific point of view. In addition, by explaining that the processing method to Korean red ginseng could increase red ginseng acidic polysaccharides (RGAP), the RGAP, PPD type ginsenosides, and arginine which is originally abundant in Korean ginseng could increase the expression of the 'heat shock proteins' as a kind of chaperone in the body, this paper presents the theory allowing the scientific interpretation of the efficacy of Korean red ginseng as an 'adaptogen' or 'medicine for anti-aging and longevity'. Lastly, through the consideration of the growing environment of American ginseng and Korean ginseng, the differences are presented.
Park, Jin-Kyu;Jin, Sung-Ha;Choi, Keum-Hee;Ko, Ji-Hun;Baek, Nam-In;Choi, Soo-Young;Cho, Sung-Woo;Choi, Kang-Ju;Nam, Ki-Yeul
BMB Reports
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v.32
no.4
/
pp.339-344
/
1999
We studied the effects of ginsenosides in immature rats based upon the previous results that ginseng has a suppressive or anticonvulsive activity. To examine the suppressive effect of ginsenosides on kainic acid-induced seizures, the severities and frequencies were observed for 4 h after injection of kainic acid (KA; i.p., 2 mg/kg b.w.) using 10-day-old male Sprague-Dawley rats ($22{\pm}2\;g$). Protopanaxadiol saponins such as ginsenoside-Rb1 (Rb1), ginsenoside-Rb2 (Rb2), ginsenoside-Rc (Rc), and ginsenoside-Rd(Rd) generally reduced the seizure activities while protopanaxatriol saponins such as ginsenoside-Rg1 (Rg1) and ginsenoside-Re (Re) rather increased stereotypic "paddling-like" movements. When vinyl-GABA (v-G) was injected together with Rb1 or Rc, KA-induced seizure severities were additionally reduced only by the injection of Rc, but not by Rb1. The level of gamma isozyme of protein kinase C (PKC-${\gamma}$) in the hippocampus increased about three times as much as that of normal rats at 4 h after KA injection. The increased level of PCK-${\gamma}$ by KA was significantly reduced to about 35% by the coinjection with v-G alone, but it was not changed by v-G together with Rb1 or Rc. The increased level of PKC-${\gamma}$ at 4 h after injection of KA was not consistent with the reduction of seizure severities between Rb1 and Rc. These results suggest that Rc and Rb1 may reduce seizure severity independent of PKC-${\gamma}$ levels, and Rc may additionally act with v-G regarding the GABA metabolism during the stage of KA-induced seizures in the immature rats.
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