Ginsenoside Rb1 and Rb2 upregulate Akt/mTOR signalingemediated muscular hypertrophy and myoblast differentiation

Ga-Yeon Go,Ayoung Jo,Dong-Wan Seo,Woo-Young Kim,Yong Kee Kim,Eui-Young So,Qian Chen,Jong-Sun Kang,Gyu-Un Bae,Sang-Jin Lee 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.3

Background: As a process of aging, skeletal muscle mass and function gradually decrease. It is reported that ginsenoside Rb1 and Rb2 play a role as AMP-activated protein kinase activator, resulting in regulating glucose homeostasis, and Rb1 reduces oxidative stress in aged skeletal muscles through activating the phosphatidylinositol 3-kinase/Akt/Nrf2 pathway. We examined the effects of Rb1 and Rb2 on differentiation of the muscle stem cells and myotube formation. Methods: C2C12 myoblasts treated with Rb1 and/or Rb2 were differentiated and induced to myotube formation, followed by immunoblotting for myogenic marker proteins, such as myosin heavy chain, MyoD, and myogenin, or immunostaining for myosin heavy chain or immunoprecipitation analysis for heterodimerization of MyoD/E-proteins. Results: Rb1 and Rb2 enhanced myoblast differentiation through accelerating MyoD/ E-protein heterodimerization and increased myotube hypertrophy, accompanied by activation of Akt/ mammalian target of rapamycin signaling. In addition, Rb1 and Rb2 induced the MyoD-mediated transdifferentiation of the rhabdomyosarcoma cells into myoblasts. Furthermore, co-treatment with Rb1 and Rb2 had synergistically enhanced myoblast differentiation through Akt activation. Conclusion: Rb1 and Rb2 upregulate myotube growth and myogenic differentiation through activating Akt/mammalian target of rapamycin signaling and inducing myogenic conversion of fibroblasts. Thus, our first finding indicates that Rb1 and Rb2 have strong potential as a helpful remedy to prevent and treat muscle atrophy, such as age-related muscular dystrophy.

Ginsenoside Rg1 from Panax ginseng enhances myoblast differentiation and myotube growth

Ga-Yeon Go,Sang-Jin Lee,Ayoung Jo,Jaecheol Lee,Dong-Wan Seo,Jong-Sun Kang,Si-Kwan Kim,Su-Nam Kim,Yong Kee Kim,Gyu-Un Bae 고려인삼학회 2017 Journal of Ginseng Research Vol.41 No.4

Background: Ginsenoside Rg1 belongs to protopanaxatriol-type ginsenosides and has diverse pharmacological activities. In this report, we investigated whether Rg1 could upregulate muscular stem cell differentiation and muscle growth. Methods: C2C12 myoblasts, MyoD-transfected 10T1/2 embryonic fibroblasts, and HEK293T cells were treated with Rg1 and differentiated for 2 d, subjected to immunoblotting, immunocytochemistry, or immunoprecipitation. Results: Rg1 activated promyogenic kinases, p38MAPK (mitogen-activated protein kinase) and Akt signaling, that in turn promote the heterodimerization with MyoD and E proteins, resulting in enhancing myogenic differentiation. Through the activation of Akt/mammalian target of rapamycin pathway, Rg1 induced myotube growth and prevented dexamethasone-induced myotube atrophy. Furthermore, Rg1 increased MyoD-dependent myogenic conversion of fibroblast. Conclusion: Rg1 upregulates promyogenic kinases, especially Akt, resulting in improvement of myoblast differentiation and myotube growth.

Ginsenoside Rb1 and Rb2 upregulate Akt/mTOR signaling – mediated muscular hypertrophy and myoblast differentiation

Ga-Yeon Go,조아영,Dong-Wan Seo,Woo-Young Kim,김용기,Eui-Young So,Qian Chen,Jong-Sun Kang,Gyu-Un Bae,Sang-Jin Lee 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.3

Background: As a process of aging, skeletal muscle mass and function gradually decrease. It is reportedthat ginsenoside Rb1 and Rb2 play a role as AMP-activated protein kinase activator, resulting in regulatingglucose homeostasis, and Rb1 reduces oxidative stress in aged skeletal muscles throughactivating the phosphatidylinositol 3-kinase/Akt/Nrf2 pathway. We examined the effects of Rb1 and Rb2on differentiation of the muscle stem cells and myotube formation. Methods: C2C12 myoblasts treated with Rb1 and/or Rb2 were differentiated and induced to myotubeformation, followed by immunoblotting for myogenic marker proteins, such as myosin heavy chain,MyoD, and myogenin, or immunostaining for myosin heavy chain or immunoprecipitation analysis forheterodimerization of MyoD/E-proteins. Results: Rb1 and Rb2 enhanced myoblast differentiation through accelerating MyoD/E-protein heterodimerization and increased myotube hypertrophy, accompanied by activation of Akt/mammalian target of rapamycin signaling. In addition, Rb1 and Rb2 induced the MyoD-mediatedtransdifferentiation of the rhabdomyosarcoma cells into myoblasts. Furthermore, co-treatment withRb1 and Rb2 had synergistically enhanced myoblast differentiation through Akt activation. Conclusion: Rb1 and Rb2 upregulate myotube growth and myogenic differentiation through activatingAkt/mammalian target of rapamycin signaling and inducing myogenic conversion of fibroblasts. Thus,our first finding indicates that Rb1 and Rb2 have strong potential as a helpful remedy to prevent andtreat muscle atrophy, such as age-related muscular dystrophy.

Ginsenoside Rg1 from Panax ginseng enhances myoblast differentiation and myotube growth

Go, Ga-Yeon,Lee, Sang-Jin,Jo, Ayoung,Lee, Jaecheol,Seo, Dong-Wan,Kang, Jong-Sun,Kim, Si-Kwan,Kim, Su-Nam,Kim, Yong Kee,Bae, Gyu-Un The Korean Society of Ginseng 2017 Journal of Ginseng Research Vol.41 No.4

Background: Ginsenoside Rg1 belongs to protopanaxatriol-type ginsenosides and has diverse pharmacological activities. In this report, we investigated whether Rg1 could upregulate muscular stem cell differentiation and muscle growth. Methods: C2C12 myoblasts, MyoD-transfected 10T1/2 embryonic fibroblasts, and HEK293T cells were treated with Rg1 and differentiated for 2 d, subjected to immunoblotting, immunocytochemistry, or immunoprecipitation. Results: Rg1 activated promyogenic kinases, p38MAPK (mitogen-activated protein kinase) and Akt signaling, that in turn promote the heterodimerization with MyoD and E proteins, resulting in enhancing myogenic differentiation. Through the activation of Akt/mammalian target of rapamycin pathway, Rg1 induced myotube growth and prevented dexamethasone-induced myotube atrophy. Furthermore, Rg1 increased MyoD-dependent myogenic conversion of fibroblast. Conclusion: Rg1 upregulates promyogenic kinases, especially Akt, resulting in improvement of myoblast differentiation and myotube growth.

Ginsenoside Rb1 and Rb2 upregulate Akt/mTOR signaling-mediated muscular hypertrophy and myoblast differentiation

Go, Ga-Yeon,Jo, Ayoung,Seo, Dong-Wan,Kim, Woo-Young,Kim, Yong Kee,So, Eui-Young,Chen, Qian,Kang, Jong-Sun,Bae, Gyu-Un,Lee, Sang-Jin The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.3

Background: As a process of aging, skeletal muscle mass and function gradually decrease. It is reported that ginsenoside Rb1 and Rb2 play a role as AMP-activated protein kinase activator, resulting in regulating glucose homeostasis, and Rb1 reduces oxidative stress in aged skeletal muscles through activating the phosphatidylinositol 3-kinase/Akt/Nrf2 pathway. We examined the effects of Rb1 and Rb2 on differentiation of the muscle stem cells and myotube formation. Methods: C2C12 myoblasts treated with Rb1 and/or Rb2 were differentiated and induced to myotube formation, followed by immunoblotting for myogenic marker proteins, such as myosin heavy chain, MyoD, and myogenin, or immunostaining for myosin heavy chain or immunoprecipitation analysis for heterodimerization of MyoD/E-proteins. Results: Rb1 and Rb2 enhanced myoblast differentiation through accelerating MyoD/E-protein heterodimerization and increased myotube hypertrophy, accompanied by activation of Akt/mammalian target of rapamycin signaling. In addition, Rb1 and Rb2 induced the MyoD-mediated transdifferentiation of the rhabdomyosarcoma cells into myoblasts. Furthermore, co-treatment with Rb1 and Rb2 had synergistically enhanced myoblast differentiation through Akt activation. Conclusion: Rb1 and Rb2 upregulate myotube growth and myogenic differentiation through activating Akt/mammalian target of rapamycin signaling and inducing myogenic conversion of fibroblasts. Thus, our first finding indicates that Rb1 and Rb2 have strong potential as a helpful remedy to prevent and treat muscle atrophy, such as age-related muscular dystrophy.

Research article Black ginseng activates Akt signaling, thereby enhancing myoblast differentiation and myotube growth

Lee, Soo-Yeon,Go, Ga-Yeon,Vuong, Tuan Anh,Kim, Jee Won,Lee, Sullim,Jo, Ayoung,An, Jun Min,Kim, Su-Nam,Seo, Dong-Wan,Kim, Jin-Seok,Kim, Yong Kee,Kang, Jong-Sun,Lee, Sang-Jin,Bae, Gyu-Un The Korean Society of Ginseng 2018 Journal of Ginseng Research Vol.42 No.1

Background: Black ginseng (BG) has greatly enhanced pharmacological activities relative to white or red ginseng. However, the effect and molecular mechanism of BG on muscle growth has not yet been examined. In this study, we investigated whether BG could regulate myoblast differentiation and myotube hypertrophy. Methods: BG-treated C2C12 myoblasts were differentiated, followed by immunoblotting for myogenic regulators, immunostaining for a muscle marker, myosin heavy chain or immunoprecipitation analysis for myogenic transcription factors. Results: BG treatment of C2C12 cells resulted in the activation of Akt, thereby enhancing hetero-dimerization of MyoD and E proteins, which in turn promoted muscle-specific gene expression and myoblast differentiation. BG-treated myoblasts formed larger multinucleated myotubes with increased diameter and thickness, accompanied by enhanced Akt/mTOR/p70S6K activation. Furthermore, the BG treatment of human rhabdomyosarcoma cells restored myogenic differentiation. Conclusion: BG enhances myoblast differentiation and myotube hypertrophy by activating Akt/mTOR/p70S6k axis. Thus, our study demonstrates that BG has promising potential to treat or prevent muscle loss related to aging or other pathological conditions, such as diabetes.

편백유(Chamaecyparis Obtusa Oil)가 DNCB로 유도된 NC/Nga생쥐의 아토피에 미치는 효과 연구

손기정,박정환,김윤하,고가연,김윤주,한지연,고기태,안택원,Son, Ki-Jeong,Park, Jung-Hwan,Kim, Youn-Ha,Go, Ga-Yeon,Kim, Yun-Joo,Han, Ji-Yeon,Ko, Ki-Tae,Ahn, Taek-Won 대한한방안이비인후피부과학회 2016 한방안이비인후피부과학회지 Vol.29 No.1

Objective : The aim of this study is to investigate the effect of Chamaecyparis Obtusa Oil to NC/Nga mice induced in Atopic dermatitis-like skin lesions by DNCB Mthods.Methods : NC/Nga mice which have been induced to Atopic dermatitis-like skin lesions by DNCB are divided into 3 groups, the one is the mice which have been spread with Chamaecyparis Obtusa Oil 2% cream, the other is which have been spread with dexamethasone (Dexa.) 0.5% on their Atopic lesion, another is the control group. then we take the blood sample of each group to measure state of the dorsal skin, the number of immunocytes. and resect the skin lesion to analyze the state of cells.Result & Conclusion : There are meaningful results of measuring the number of IgE, IL-4, IFN-γ and the state of the skin lesion and cells in the group with hamaecyparis Obtusa Oil 2% cream in comparison with the other groups.

Peroxisome proliferator-activated receptor β/δ (PPARβ/δ) activates promyogenic signaling pathways, thereby promoting myoblast differentiation

Lee, Sang-Jin,Go, Ga-Yeon,Yoo, Miran,Kim, Yong Kee,Seo, Dong-Wan,Kang, Jong-Sun,Bae, Gyu-Un Elsevier 2016 Biochemical and biophysical research communication Vol.470 No.1

<P><B>Abstract</B></P> <P>Peroxisome proliferator-activated receptor β/δ (PPARβ/δ) regulates postnatal myogenesis by alleviating myostatin activity, but the molecular mechanisms by which it regulates myogenesis are not fully understood. In this study, we investigate molecular mechanisms of PPARβ/δ in myoblast differentiation. C2C12 myoblasts treated with a PPARβ/δ agonist, GW0742 exhibit enhanced myotube formation and muscle-specific gene expression. GW0742 treatment dramatically activates promyogenic kinases, p38MAPK and Akt, in a dose-dependent manner. GW0742-stimulated myoblast differentiation is mediated by p38MAPK and Akt, since it failed to restore myoblast differentiation repressed by inhibition of p38MAPK and Akt. In addition, GW0742 treatment enhances MyoD-reporter activities. Consistently, overexpression of PPARβ/δ enhances myoblast differentiation accompanied by elevated activation of p38MAPK and Akt. Collectively, these results suggest that PPARβ/δ enhances myoblast differentiation through activation of promyogenic signaling pathways.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A PPARβ/δ agonist, GW0742 promotes myoblast differentiation. </LI> <LI> GW0742 activates both p38MAPK and Akt activation in myogenic differentiation. </LI> <LI> GW0742 enhances MyoD activity for myogenic differentiation. </LI> <LI> Overexpression of PPARβ/δ enhances myoblast differentiation via activating promyogenic signaling pathways. </LI> <LI> This is the first finding for agonistic mechanism of PPARβ/δ in myogenesis. </LI> </UL> </P>

MPTP로 유도된 Parkinson's disease 동물 모델에서 열다한소탕 가감방 (MYH)의 신경 세포 보호 효과

고가연,김윤하,안택원,Go, Ga-Yeon,Kim, Yoon-Ha,Ahn, Taek-Won 사상체질의학회 2015 사상체질의학회지 Vol.27 No.2

Objectives To evaluate the neuroprotective effects of modified Yuldahanso-tang (MYH) in a Parkinson's disease mouse model. Methods 1) Four groups (each of 8 rats per group) were used in this study. 2) The neuroprotective effect of MYH was examined in a Parkinson's disease mouse model. C57BL/6 mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day), intraperitoneal (i.p.) for 5 days. 3) The brains of 2 mice per group were removed and frozen at $-20^{\circ}C$, and the striatum-substantia nigra part was seperated. The protein volume was measured by Bradford method following Bio-Rad protein analyzing kit. Using mouse/Rat Dopamine ELISA Assay Kit. 4) The brains of 2 mice per group were separated and removed. TH-immunohistochemical was examined in the MPTP-induced Parkinson's disease mice to evaluate the neuroprotective effects of MYH on ST and SNpc. 5) Two mice out of each group were anesthetized and skulls were opened from occipital to frontal direction to take out the brains. The brains added TTC solution for 20 minutes for staining. 6) The water tank used for morris water maze test was filled with $28^{\circ}C$ water, and a round platform of 10cm in diameter was installed for mice to step on. The study was carried out once a day within 30 seconds, keep exercising to step on the platform in the pool. 7) The brains of two mice out of each group were fixed in 10% formaldehyde solution and paraphillin substance was infiltrated. They were fragmented by microtome, and observed under an optical microscope after Hematoxylin & Eosin staining. 8) A round acrylic cylinder with its upper side open was filled with clean water and depressive mouse models were forced to swim for 15 minutes. After 24 hours the animals were put in the same equipment for 5 minutes and were forced to swim. 9) The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of Neurotransmitters in MPTP-MYH group. Results 1) MYH possess Dopamine cell protective effect on MPTP-induced injury in striatum and substantia nigra pars compacta. 2) MYH inhibits the loss of tyrosine hydroxylase-immunoreacitive (TH-IR) cells in the striatum and substantia nigra pars compacta on MPTP-induced injury in C57BL/6 mice. 3) MYH possesses improvement effect on MPTP-induced memory deterioration in C57BL/6 mice through the reduction of prolongated Sort of lost time by MPTP injection using the Morris water maze test. 4) MYH possesses hippocampal neuron protective effect on MPTP-induced injury in C57BL/6 mice. 5) MYH possesses improvement effect on MPTP-induced motor behaviour deficits and depression in C57BL/6 mice through the reduction of prolongated losing motion by MPTP injection using the Forced swimming test. 6) MYH increases serotonin product amount on MPTP-induced injury in C57BL/6 mice. Conclusions This experiment suggests that the neuroprotective effect of MYH is mediated by the increase in Dopamin, TH-ir cell, Hippocampus and Serotonin. Furthermore, MYH essential oil may serve as a potential preventive or therapeutic agent regarding Parkinson's disease.

MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)로 유도된 Parkinson's Disease 동물 모델을 이용한 향사양위탕의 신경 세포 보호 효과

고가연,김윤희,안택원,Go, Ga-Yeon,Kim, Yun-Hee,Ahn, Taek-Won 사상체질의학회 2014 사상체질의학회지 Vol.26 No.2

Objectives To evaluate the neuroprotective effects of Hyangsayangwi-tang (HY), a Korean traditional medicinal prescription in a Parkinson's disease mouse model. Methods Four groups(each of 10 mouse per group) were used in this study. The neuroprotective effect of HY was examined in a Parkinson's disease mouse model. C57BL/6 mouse treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30mg/kg/day), intraperitoneal (i.p.) for 5 days. Slow behavioral responses and memory disorder is the major clinical symptoms of PD. In order to investigate the effect of HY on recovery of behavioral deficits and memory, we examined the motor function and memory by using Morris water maze and Forced swimming test. Ischemic mouse brain stained with TTC(2,3,5 triphenyl tetrazolium chloride) in the MPTP-induced Parkinson's disease to find out ischemia and tissue damage in mouse. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of neurotransmitters in MPTP-HY group. To measure the amount of dopamine in mice brain, striatum-substantia nigra, was examined by Bradford assay. Immunohistochemistry was examined in the MPTP-induced Parkinson's disease (PD) mouse to evaluate the neuroprotective effects of Hyangsayangwi-tang on hippocampal lesion, ST and SNpc. Results and Conclusions Hyangsayangwi-tang (HY) prevents MPTP-induced loss of serotonin, hippocampus and TH-ir cell.