Effects of $\alpha$-lipoic acid on cell proliferation and apoptosis in MDA-MB-231 human breast cells

Na, Mi-Hee,Seo, Eun-Young,Kim, Woo-Kyoung The Korean Nutrition Society 2009 Nutrition Research and Practice Vol.2 No.3

The role that antioxidants play in the process of carcinogenesis has recently gained considerable attention. $\alpha$-Lipoic acid, a naturally occurring disulfide molecule, is a powerful antioxidant that reportedly exerts beneficial effects in patients with advanced cancer by reducing the level of reactive oxygen species and increasing glutathione peroxidase activity. In this study, we examined changes in the protein and mRNA expression associated with cell proliferation and apoptosis in MDA-MB-231 breast cancer cultured in the presence of various concentrations (0, 250, 500, and 1000 ${\mu}mol/L$) of $\alpha$-lipoic acid. The results revealed that $\alpha$-lipoic acid inhibited the growth of breast cancer cells in a dose-independent manner (P < 0.05). Additionally, $ErbB_2$ and $ErbB_3$ protein and mRNA expressions were significantly decreased in a dose-dependent manner in response to $\alpha$-lipoic acid (P < 0.05). Furthermore, the protein expression of phosphorylated Akt (p-Akt) levels and total Akt, and the mRNA expression of Akt were decreased dose-dependently in cells that were treated with $\alpha$-lipoic acid (P < 0.05). Bcl-2 protein and mRNA expressions were also decreased in cells that were treated with $\alpha$-lipoic acid (P < 0.05). However, Bax protein and mRNA expressions were increased in cells treated with $\alpha$-lipoic acid (P < 0.05). Finally, caspase-3 activity was significantly increased in a dose-dependent manner in cells treated with $\alpha$-lipoic acid (P < 0.05). In conclusion, we demonstrated that $\alpha$-lipoic acid inhibits cell proliferation and induces apoptosis in MDA-MB-231 breast cancer cell lines.

α-Lipoic acid의 희석용매, 처리농도, 처리시간에 따른 3T3-L1 지방세포 성장에 미치는 영향

서은영,Seo, Eunyoung 한국식생활문화학회 2018 韓國食生活文化學會誌 Vol.33 No.5

Purpose: This study examined the effects of ${\alpha}$-lipoic acid in diluted solvents on cell growth in 3T3-L1 cells according to the treated concentration and times. Methods: Adipocyte 3T3-L1 cell were cultured. Confluent cells underwent starvation with SFM for 1 day and then were cultured in a medium containing various concentrations 0, 100, 200, and $400{\mu}mol/L$ of ${\alpha}$-lipoic acid. The cell viability was measured using the EZ Cytox assay kit. In addition, the effect of ${\alpha}$-lipoic acid of diluted solvents on the cell growth in 3T3-L1cells was examined according to the treated concentration and times. Results: The ${\alpha}$-lipoic acid diluted ethanol inhibited cell proliferation in a dose and time dependent manner. The ${\alpha}$-lipoic acid diluted ethanol induced adipocyte 3T3-L1 cells proliferation with an adipocyte inducer. In addition, ${\alpha}$-lipoic acid inhibited adipocyte 3T3-L1 growth in a dose and time dependent manner (p<0.05). Conclusion: This study showed that a treatment with ${\alpha}$-lipoic acid diluted ethanol inhibits cell growth of, adipocyte 3T3-L1 cells induced with an adipocyte inducer, ($200{\mu}mol/L$ of ${\alpha}$-lipoic acid) treated for 48 hr.

Effects of ${\alpha}$-lipoic acid and L-carnosine supplementation on antioxidant activities and lipid profiles in rats

Kim, Mi-Young,Kim, Eun-Jin,Kim, Young-Nam,Choi, Chang-Sun,Lee, Bog-Hieu The Korean Nutrition Society 2011 Nutrition Research and Practice Vol. No.

${\alpha}$-Lipoic acid and L-carnosine are powerful antioxidants and are often used as a health supplement and as an ergogenic aid. The objective of this study was to investigate the effects of ${\alpha}$-lipoic acid and/or L-carnosine supplementation on antioxidant activity in serum, skin, and liver of rats and blood lipid profiles for 6 weeks. Four treatment groups received diets containing regular rat chow diet (control, CON), 0.5% ${\alpha}$-lipoic acid (ALA), 0.25% ${\alpha}$-lipoic acid+0.25% L-carnosine (ALA+LC), or 0.5% L-carnosine (LC). Superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and lipid peroxidation products, malondialdehyde (MDA) concentrations, were analyzed in serum, skin, and liver. Blood lipid profiles were measured, including triglycerides (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C). Skin and liver SOD activities of the ALA and LC groups were higher than those of the CON group (P<0.05), but serum SOD activity was higher only in the LC group compared to that in the CON group (P<0.05). Additionally, only liver GSH-Px activity in the LC group was higher than that of the CON and the other groups. Serum and skin MDA levels in the ALA and LC groups were lower than those in the CON group (P<0.05). Serum TG and TC in the ALA and ALA+LC groups were lower than those in the CON and LC groups (P<0.05). The HDL-C level in the LC group was higher than that in any other group (P<0.05). LDL-C level was lower in the ALA+LC and LC groups than that in the CON group (P<0.05). Thus, ${\alpha}$-lipoic acid and L-carnosine supplementation increased antioxidant activity, decreased lipid peroxidation in the serum, liver, and skin of rats and positively modified blood lipid profiles.

Alpha-Lipoic Acid Inhibits Glycogen Synthesis and Modifies Glucose Metabolism and Signaling Pathways in Soleus Muscles from Healthy Rats

Madar, Zecharia,Stark, Aliza H.,Ilan, Erez,Timar, Bracha,Borenshtein, Diana The Korean Society of Food Science and Nutrition 2002 Preventive Nutrition and Food Science Vol.7 No.2

Alpha-lipoic acid is a known hypoglycemic agent that may be useful in the treatment of diabetes. The objective of this study was to investigate the fate of glucose in isolated muscles incubated with lipoic acid by determining its direct effects on specific metabolic and signaling pathways. Soleus muscles from healthy rats were incubated with lipoic acid in the absence or presence of insulin. Glucose transport, glycogen synthesis, glucose oxidation and lipid synthesis were determined and affects on major pathways associated with insulin signaling were evaluated. Glucose transport was not significantly altered by the addition of lipoic acid to the incubation medium. However, lipoic acid decreased glycogen synthesis in comparison to controls. Glucose oxidation was moderately increased while de-novo lipid synthesis from glucose was inhibited. Wortmannin repressed insulin stimulation of glucose incorporation into glycogen, an effect that was augmented by the combined treatment of wortmannin and lipoic acid. Basal and insulin-stimulated serine phosphorylation of Akt was not changed by the addition of lipoic acid to the incubation medium. These data show that in this in vitro model, lipoic acid did not significantly affect glucose uptake but dramatically modified pathways of glucose metabolism within muscle tissue.

α-lipoic acid protects testis and epididymis against linuron-induced oxidative toxicity in adult rats

Prathima P.,Venkaiah K.,Daveedu T.,Pavani R.,Sukeerthi S.,Gopinath M.,Sainath Sri Bhashaym 한국독성학회 2020 Toxicological Research Vol.36 No.4

Linuron is well known for its antiandrogenic property. However, the effects of linuron on testicular and epididymal pro- and antioxidant status are not well defined. On the other hand, α-lipoic acid is well known as universal antioxidant. Therefore, the purpose of this study was twofold: firstly to investigate whether linuron exposure alters antioxidant status in the testis and epididymis of rats and if so, whether the supplementation of α-lipoic acid mitigates linuron-induced oxidative toxicity in rats. To address this question, α-lipoic acid at a dose of 70 mg/Kg body weight (three times a week) was administered to linuron exposed rats (10 or 50 mg/Kg body weight, every alternate day over a period of 60 days), and the selected reproductive endpoints were analyzed after 60 days. Respective controls were maintained in parallel. Linuron at selected doses reduced testicular daily sperm count, and epididymal sperm count, sperm motility, sperm viability, and number of tail coiled sperm, reduced activity levels of 3β- and 17β-hydroxysteroid dehydrogenases, decreased expression levels of StAR mRNA, inhibition of testosterone levels, and elevated levels of testicular cholesterol in rats over controls. Linuron intoxication deteriorated the structural integrity of testis and epididymis associated with reduced the reproductive performance over controls. Conversely, α-lipoic acid supplementation enhanced sperm quality and improved the testosterone synthesis pathway in linuron exposed rats over its respective control. Administration of α-lipoic acid restored inhibition of testicular and epididymal enzymatic (superoxide dismutase, catalase, glutathione reductase, glutathione peroxidise) and non-enzymatic (glutathione content), increased lipid peroxidation and protein carbonyl content produced by linuron in rats. α-lipoic acid supplementation inhibited the expression levels of testicular caspase-3 mRNA levels and also its activity in linuron treated rats. To summate, α-lipoic acid-induced protection of reproductive health in linuron treated rats could be attributed to its antioxidant, and steroidogenic properties.

흰쥐 후지근 피판에서 허혈-재순환 손상시 pERK1/2 발현에 대한 ${\alpha}-lipoic$ Acid의 효과

송정훈,김민선,박병림,박한수,채정룡,이혜미,나영천,Song, Jeong-Hoon,Kim, Min-Sun,Park, Byung-Rim,Park, Han-Su,Chae, Jeong-Ryong,Lee, Hye-Me,Na, Young-Cheon 대한미세수술학회 2005 Archives of reconstructive microsurgery Vol.14 No.2

Purpose: This study was to evaluate the effect of ${\alpha}-lipoic$ acid, a potent free radical scavenger, on the expression of active form of extracellular signal-regulated kinase (pERK1/2) proteins from hindlimb muscles of rats following ischemia-reperfusion injury. Material and methods: 64 health, $280{\sim}350\;g$ weighted Sprague-Dawley male rats were used. In order to make a muscle flap, the gastrocnemius (GC) and soleus (SOL) muscles were dissected and elevated. The popliteal artery was occluded for 4hours and reperfused for 10 minutes, 30 minutes, 1 hour, 2 hours and 4 hours, respectively. Results: The ischemia by occlusion of the popliteal artery itself caused a minimal change in expression of phosphorylated form of proteins observed in hindlimb muscle. In contrast, after 4 hours of ischemia, immunoreactivity for pERK1/2 in the GC muscle showed dual peaks at 10 minutes and 4 hours after reperfusion. In ${\alpha}-lipoic$ acid treated group, the expression of pERK1/2 was increased significantly compared to I/R-only group. Conclusion: These results suggest that ${\alpha}-lipoic$ acid may protect I/R injury of the skeletal muscle through free radical scavening and activation of intracellular pERK1/2 expression.

Alpha Lipoic acid와 연관된 인슐린자가면역증후군

장형진 ( Hyoung Jin Chang ),최현숙 ( Hyun Sook Choi ),박미연 ( Mi Youn Park ),임성민 ( Sung Min Leem ),장이선 ( Yi Sun Jang ),박강서 ( Kang Seo Park ),이종민 ( Jong Min Lee ) 대한내과학회 2009 대한내과학회지 Vol.76 No.5

Insulin autoimmune syndrome is characterized by spontaneous hypoglycemia, high concentrations of serum immunoreactive insulin, and the presence of autoantibodies to insulin without previous insulin injection. A 71-year-old woman with diabetes, who had been treated with oral hypoglycemic agents, suffered from frequent hypoglycemia. High insulin levels and the presence of insulin autoantibodies were shown, so insulin autoimmune syndrome was diagnosed. Drugs containing sulfhydryl groups play an important role in the pathogenesis of insulin autoimmune syndrome. Alpha-lipoic acid, which contains a sulfhydryl group, was administered before the onset of hypoglycemia. The patient`s human leukocyte antigen (HLA) type was HLA-Cw4 and DRB1*0406. There is a strong correlation between HLA-DRB1*0406 and insulin autoimmune syndrome. The patient was treated with prednisolone and has not had a hypoglycemic attack since. We report a case of insulin autoimmune syndrome, possibly associated with alpha-lipoic acid. (Korean J Med 76:600-604, 2009)

Protective Efficacy of Alpha-lipoic Acid against AflatoxinB1-induced Oxidative Damage in the Liver

Li, Y.,Ma, Q.G.,Zhao, L.H.,Guo, Y.Q.,Duan, G.X.,Zhang, J.Y.,Ji, C. Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.6

Alpha-lipoic acid (${\alpha}$-LA) is not only involved in energy metabolism, but is also a powerful antioxidant that can protect against hepatic oxidative stress induced by some drugs, toxins, or under various physiological and pathophysiological conditions. Here, we investigated the effect of ${\alpha}$-LA against liver oxidative damage in broilers exposed to aflatoxin $B_1$ ($AFB_1$). Birds were randomly divided into four groups and assigned different diets: basal diet, 300 mg/kg ${\alpha}$-LA supplementation in basal diet, diet containing 74 ${\mu}g/kg$ $AFB_1$, and 300 mg/kg ${\alpha}$-LA supplementation in diet containing 74 ${\mu}g/kg$ $AFB_1$, for 3 weeks. The results revealed that the addition of 300 mg/kg ${\alpha}$-LA protected against the liver function damage of broilers induced by chronic low dose of $AFB_1$ as estimated by a significant (p<0.05) change in levels of plasma total protein, albumin, alkaline phosphatase and the activities of liver glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase. The histopathological analysis also showed that liver tissues were injured in the $AFB_1$ diet, but this effect was alleviated by the addition of 300 mg/kg ${\alpha}$-LA. Additionally, $AFB_1$ induced a profound elevation of oxidative stress in birds, as indicated by an increase in malondialdehyde level, a decrease in glutathione peroxidase activity and a depletion of the glutathione content in the liver. All of these negative effects were inhibited by treatment with ${\alpha}$-LA. Our results suggest that the inhibition of $AFB_1$-induced excess production of lipid peroxides and the maintenance of intracellular antioxidant status may play important roles in the protective effects of ${\alpha}$-LA against $AFB_1$-induced oxidative damage in the liver.

Cardioprotective Effects of Alpha-Lipoic Acid on Myocardial Reperfusion Injury: Suppression of Reactive Oxygen Species Generation and Activation of Mitogen-Activated Protein Kinase

오석규,윤경호,유남진,김남호,김민선,박병림,정진원 대한심장학회 2009 Korean Circulation Journal Vol.39 No.9

Background and Objectives: Reactive oxygen species (ROS) and mitogen-activated protein (MAP) kinase play an important role in the development of myocardial reperfusion injury. In this study, we examined whether treatment with alpha-lipoic acid (ALA) before reperfusion could prevent myocardial reperfusion injury in vivo. Materials and Methods: Sprague-Dawley rats were subjected to a 45-minute left anterior descending coronary artery ligation followed by 45- or 10-minute reperfusion. ALA was administered 10 minutes prior to reperfusion. The infarct size ratio of the infarct area to the ischemic area at risk, was measured based on 10, 25, 50, and 100 mg/kg of ALA, with propidium iodide (PI) fluorescence. Apoptosis was evaluated by TdT-mediated dUDP nick end labeling (TUNEL) staining. The generation of intracellular ROS was evaluated using the fluorogenic probe, dichlorodihydrofluorescein diacetate (CM-H2DCFDA). Western blot analysis was performed for MAP kinase (pERK 1/2 and pJNK 1/2) activity. Results: The infarct size, according to ALA dose, was significantly suppressed 29.1% with ALA 25 mg/kg (p<0.0001), 41.5% with 50 mg/kg (p<0.05), and 41.4% with 100 mg/kg (p<0.05) compared to the controls (54.3%). However, the results were not significantly different with 47.2% of the ALA 10 mg/kg (p=0.192). A few apoptotic nucleoli were detected in the ALA 25 mg/kg group, but were frequently detected in the control group. The ROS generation was significantly suppressed (p<0.0001), the activity of pERK 1/2 was significantly increased (p<0.05) and the activity of pJNK 1/2 was significantly decreased (p<0.05) in the ALA 25 mg/kg group compared to the controls. Conclusion: The results of this study suggested that adequate doses of ALA before reperfusion was effective for the prevention of myocardial reperfusion injury in vivo. This cardioprotective activity of ALA might be associated with an anti-apoptotic effect of ALA via suppression of ROS generation, increase of pERK 1/2 and decrease of pJNK 1/2 activity. Background and Objectives: Reactive oxygen species (ROS) and mitogen-activated protein (MAP) kinase play an important role in the development of myocardial reperfusion injury. In this study, we examined whether treatment with alpha-lipoic acid (ALA) before reperfusion could prevent myocardial reperfusion injury in vivo. Materials and Methods: Sprague-Dawley rats were subjected to a 45-minute left anterior descending coronary artery ligation followed by 45- or 10-minute reperfusion. ALA was administered 10 minutes prior to reperfusion. The infarct size ratio of the infarct area to the ischemic area at risk, was measured based on 10, 25, 50, and 100 mg/kg of ALA, with propidium iodide (PI) fluorescence. Apoptosis was evaluated by TdT-mediated dUDP nick end labeling (TUNEL) staining. The generation of intracellular ROS was evaluated using the fluorogenic probe, dichlorodihydrofluorescein diacetate (CM-H2DCFDA). Western blot analysis was performed for MAP kinase (pERK 1/2 and pJNK 1/2) activity. Results: The infarct size, according to ALA dose, was significantly suppressed 29.1% with ALA 25 mg/kg (p<0.0001), 41.5% with 50 mg/kg (p<0.05), and 41.4% with 100 mg/kg (p<0.05) compared to the controls (54.3%). However, the results were not significantly different with 47.2% of the ALA 10 mg/kg (p=0.192). A few apoptotic nucleoli were detected in the ALA 25 mg/kg group, but were frequently detected in the control group. The ROS generation was significantly suppressed (p<0.0001), the activity of pERK 1/2 was significantly increased (p<0.05) and the activity of pJNK 1/2 was significantly decreased (p<0.05) in the ALA 25 mg/kg group compared to the controls. Conclusion: The results of this study suggested that adequate doses of ALA before reperfusion was effective for the prevention of myocardial reperfusion injury in vivo. This cardioprotective activity of ALA might be associated with an anti-apoptotic effect of ALA via suppression of ROS generation, increase of pERK 1/2 and decrease of pJNK 1/2 activity.

리포익산을 함유한 PNIPAM 하이드로젤의 제조

윤혜리(Hye Ri Yoon),이종휘(Jong Hwi Lee) 한국고분자학회 2012 폴리머 Vol.36 No.4

Poly(N-isopropylacrylamide)(PNIPAM)는 체온과 비슷한 온도에서 부피상 변화 혹은 온도 감응성 팽윤 거동의 특성을 보여 약물전달시스템에서 중요하게 연구되고 있다. 그러나 PNIPAM의 친수성 특징 때문에 소수성 약물을 그 네트워크 안에 고르게 넣기는 쉽지 않다. 항산화제인 알파 리포익산은 개환중합으로 고분자화(polylipoic acid, PLA) 될 수는 있으나, 분자량이 낮고 분해되기 쉬워 고분자 재료로 사용되기에는 어려움이 많다. 이러한 결점들을 극복하기 위해 소수성 활성성분인 알파 리포익산을 NIPAM과 반응시켜 안정적인 하이드로젤을 만들었다. 단순한 혼합과 가열에 의한 라디칼 반응으로 하이드로젤(PNIPAM/PLA)이 성공적으로 만들어졌고, 이를 DSC, FTIR, Raman spectroscopy를 통해서 확인하였다. PNIPAM/PLA는 온도 감응성 특성을 보여주며, 리포익산의 함량이 증가 할수록 부피팽창 정도는 감소하였다. 이러한 하이드로젤을 사용하여 PNIPAM에 소수성 약물을 쉽게 담지 할 수 있고 리포익산은 항산화제로 효과가 있어, 본하이드로젤은 최종 약물전달 체로서도 유용하다. Poly(N-isopropylacrylamide) (PNIPAM) hydrogel has been studied as an important drug delivery system due to its volume transition or temperature-responsive swelling properties, whose phase separation temperature is similar to the body temperature. However, because of hydrophilic PNIPAM, hydrophobic drugs are difficult to be uniformly loaded in the networks. Antioxidant alpha-lipoic acid (LA) can be prepared as a polymer (polylipoic acid, PLA) by ring opening polymerization, which is hardly developed as a material due to its low molecular weight and easy depolymerization. To overcome this limitation, a hydrophobic active ingredient, LA was reacted with NIPAM into stable hydrogels. Simple thermal radical reaction successfully resulted in a hydrogel (PNIPAM/PLA), which was confirmed by DSC, FTIR, and Raman spectroscopy. The PNIPAM/PLA showed temperature-responsive properties, and their volume swelling decreased with an increase in lipoic acid content. These hydrogels can carry hydrophobic drugs with PNIPAM and the hydrogels could be useful as final drug delivery systems having lipoic acid as an antioxidant.