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C2C12 myotube의 산화적 손상에 대한 혼합 한약재 추출물(HME)의 Akt/FoxO3 신호 조절을 통한 보호 효과
김소영,최문열,이은탁,추성태,김미려 대한본초학회 2022 大韓本草學會誌 Vol.37 No.4
Objectives : In this study, we investigated the synergistic protective effects of medicinal herbal mixture (HME) including Mori Ramulus (MR), Acanthopanacis Cortex (AC), Eucommiae Cortex (EC), and Black soybean (BS) in C2C12 cells, mouse myoblasts. Methods : Effects of HME on cell viability of C2C12 myoblasts were monitored by MTT assay. Anti-atrophic activity of HME was determined in myoblasts and myotubes under oxidative stress by H2O2. C2C12 myoblasts were differentiated into myotubes in a medium containing 2% horse serum for 6 days. After that, we measured that expression of MyoD and myogenine, the myogenic regulatory factors, to identify the mechanism of inhibiting muscle atophy after HME treatment. In addition, suppression of phosphorylation of Akt, FoxO3a and MARF-1, transcription factors of degradation proteins were analyzed via western blotting. Results : As a result of MTT, HME there was no show cytotoxicity up to a concentration of 1 mg/ml. The cytoprotective effects on oxidative stressed myoblast and myotube was better in HME extract than those of MR, AC, EU, and BS, respectively. HME treatment in Myotube induced by oxidative stress after H2O2 treatment increased Myo D, Myogenine activation, and Akt, FoxO3a phosphorylation and decreased expression of MuRF-1. As the results, HME has synergistic effects on protection against proteolysis of C2C12 myotubes through activation of the Akt signaling pathway under oxidative stress. Conclusions : These results suggest that HME may also be useful as a preventing and treating material for skeletal muscle atrophy caused by age-related diseases.
김유진 ( Yoo-jin Kim ),김소영 ( So Young Kim ),정미진 ( Mi Jin Jeong ),이은탁 ( Un-tak Lee ),추성태 ( Sung-tae Choo ),윤석나 ( Seok Na Youn ),김미려 ( Mi Ryeo Kim ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.3
Objectives : Butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) were well known as anti-oxidant, but they were limited to use because of toxicity. So, many studies are being done to develope natural anti-oxidant. Total phenolic and flavonoid contents along with total antioxidant capacity of the ethanolic extract of Plantaginis Herba (PH) were evaluated to explore the reliable and potential sources of novel natural antioxidants. Methods : Total polyphenol contents and total flavonoid contents in PH ethanol extract were determined by colorimetric method. And DPPH(1.1-diphenyl-2-picrylhydrazyl), ABTS(2'-azino-bis (3-ethylbenzothiazoline-6-Surfonicacid)) free radical scavenging capacity and reducing power inhibition activities of PH ethanol extract were measured at 100, 500, 1000, 5000 ㎍/㎖ concentrations by spectrometric assay. Results : The total polyphenol contents and total flavonoid contents of the extract were 161.99 ㎎/g, 144.05 ㎎/g, respectively. Also, DPPH, ABTS free radical scavenging capacity and reducing power of PH ethanol extract in treated concentrations (100, 500, 1000, 5000 ㎍/㎖) increased dose dependently. In particular, DPPH free radical scavenging capacity of PH ethanol extract from 500 ㎍/㎖ was significantly increased compared to positive control (BHA). ABTS free radical scavenging capacity of PH ethanol extract from 1000 ㎍/㎖ was significantly higher than BHA. Also, reducing power showed that PH ethanol extract from 500 ㎍/㎖ was significantly increased compared to BHA. Conclusions : These results suggest that PH ethanol extract has effects to scavenge free radicals, thus PH has potential and applicable benefits for development of materials and products to have anti-oxidation functions.
차전초 잎의 물 추출물이 고지방식이로 비만 유도된 C57BL/6 생쥐의 체지방 감소에 미치는 영향
김소영 ( So Young Kim ),정미진 ( Mi Jin Jeong ),김유진 ( Yoo Jin Kim ),이은탁 ( Un-tak Lee ),추성태 ( Sung-tae Choo ),김한혁 ( Han-hyuk Kim ),김미려 ( Mi Ryeo Kim ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.2
Objective : Plantago asiatica L. has been used for many years in Korea, China, and Japan for the treatment of many diseases such as anti-inflammatory, anti-asthma and diuretic. But the anti-obesity effect of Plantaginis asiaticae Folium has not been studied yet. Thus, this study aimed to demonstrate the anti-obesity efficacy of Plantaginis asiaticae Folium water extract (PAF) in high fat diet-induced obesity animal model. Method : Male, five-weeks old C57BL/6 mice were divided into 5 groups; ND (normal diet + 0.9% saline), HFD (highfat diet + 0.9% saline), PC (high-fat diet + Garcinia cambogia 500 ㎎/㎏), PAF 100, 300 (high-fat diet + PAF 100, 300 ㎎/㎏. Treatments were performed daily for 6 weeks per os. We evaluated the changes of body weight, white adipose tissues weight, hepatic and fecal lipids level, pancreatic lipase activity and lipid profiles in plasma. Result : Body weights, adipose tissue weights and FER were significantly lower in mice fed the high-fat diet with PAF treatment than in mice fed the high-fat diet alone. In plasma, TG, TC, LDL-C, ATS and ALT levels significantly decreased in PAF groups compared with HFD group. Also, hepatic triglyceride (TG) levels decreased and fecal TG increased in PAF groups compared to HFD group. In addition, the activity of pancreatic lipase was decreased in the PAF groups compared to that of HFD group. Conclusion: These results suggest PAF extract is effective in preventing and improvement obesity, so it might be a potential and safe material for anti-obesity agent of weight control or functional supplements.
차전초 잎의 물 추출물이 고지방식이로 비만이 유도된 C57BL/6 생쥐에서 지질대사 단백질 발현에 미치는 영향
김소영 ( So Young Kim ),정미진 ( Mi Jin Jeong ),김유진 ( Yoo Jin Kim ),이은탁 ( Un-tak Lee ),추성태 ( Sung-tae Choo ),김미려 ( Mi Ryeo Kim ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.3
Objective : Previous studies showed that water extract of Plantago asiatica (Plantaginis asiaticae Folium, PAF) significantly controlled in body weights, adipose tissue weights and blood lipid profiles in obese C57BL/6 mice. To investigate the mechanism of anti-obesity action of PAF, expressions of obesity-related proteins were identified such as p-AMPK and p-ACC in hypothalamus, UCP-1 in brown adipose tissue, p-AMPK, p-ACC, SREBP-1c, PPARγ, HMGCR and CPT-1 in liver. Method : Five-weeks old male C57BL/6 mice were divided into 5 groups; ND (normal diet + 0.9% saline), HFD (high-fat diet + 0.9% saline), PC (high-fat diet+Garcinia cambogia 500 ㎎/㎏), PAF 100 and 300 (high-fat diet + PAF 100 or 300 ㎎/㎏). PAF was treated orally for 6 weeks. The protein expression of AMPK, p-AMPK, ACC, p-ACC, PPARγ, SREBP-1c, HMGCR, CPT-1 and UCP-1 were identified by expression levels of proteins through western blot analysis. Result : The results showed that protein expressions on hypothalamic p-AMPK and p-ACC did not differ between the HFD and PAF groups. In addition, PAF did not affect the increase of UCP-1 in brown adipose tissue. The protein expression levels of hepatic p-AMPK, p-ACC and CPT-1 increased in PAF groups compared to HFD group. And those of PPARγ, SREBP-1c and HMG-CoA decreased in PAF groups compared to HFD group. Conclusion : These results suggest that the PAF administration induce weight loss via inhibition of lipid metabolism-related protein expressions in hepatic tissues. Therefore, PAF could be used as a potent material of anti-obesity products for prevention and treatment of obesity.
추출조건에 따른 차전초 추출물이 3T3-L1 세포의 지방 축적에 미치는 영향
정미진 ( Mi-jin Jeong ),김소영 ( So-young Kim ),김유진 ( Yoo-jin Kim ),이은탁 ( Eun-tag Lee ),추성태 ( Sung-tae Choo ),김한혁 ( Han-hyuk Kim ),김미려 ( Mi Ryeo Kim ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.2
Objectives : Plantaginis asiaticae Folium (PA) has been widely used in Korean medicine for treatment of liver disease, stomach troubles and inflammation. We investigated the effect of PA on lipid accumulation in 3T3-L1 according to extraction conditions. Methods : The MTT assay was employed to evaluate the cytotoxicity of PA extracted by different solvents (water, 50% ethanol, and 95% ethanol) on 3T3-L1 preadipocytes. Oil red O staining was used to identify intracellular lipid accumulation in 3T3-L1. 3T3-L1 adipocytes were treated with PA at concentration ranging in 0.1, 0.2 and 0.4 ㎎/㎖. PA was extracted by different extraction conditions such as extraction solvents, extraction time, and extraction temperature. In addition, UPLC analysis was used for determination of candidates of active ingredients in PA. Results : 3T3-L1 preadipocytes were treated with PA extracted by different solvents (water, 50% ethanol, and 95% ethanol) and there was no cytotoxicity. Oil red O staining was employed to identify the effect of PA on lipid accumulation in 3T3-L1. In the present study, PA water extraction at 70℃ for 6 hours decreased greatly in lipid accumulation. The range of concentrations was 0.1, 0.2 and 0.4 ㎎/㎖. Concentration at 0.2 ㎎/㎖ was the most effective one among them. Candidates of active ingredients in PA were shown plantamajoside and acteoside through UPLC. Conclusions : These results suggest that the effect of PA water extraction at 70℃ on lipid accumulation in 3T3-L1 is superior to other extraction conditions. We suppose that plantamajoside and acteoside may be candidates of active ingredients in PA.