자연과학편 : 운동과 Hesperetin이 고콜레스테롤식이를 섭취한 흰쥐의 혈중 지질성분에 미치는 영향

이수천(SooChunLee),전혜린(HyeRinJeon),이윤경(YounKyungLee),민진아(JinAhMin),서효빈(HyoBinSeo),류승필(SungPilRyu),윤신중(ShinJoongYoun) 한국체육학회 2007 한국체육학회지 Vol.46 No.5

본 연구는 고콜레스테롤식이를 섭취하는 흰쥐를 대상으로 6주간의 트레드밀 운동과 최근 혈중지질저하 등 유익한 효능을 가진 주요 감귤류 플라보노이드인 hesperetin의 처치가 혈중 지질성분에 미치는 영향을 분석하였다. SD 수컷 흰쥐 32마리를 대상으로 대조군, 운동군, hesperetin 처치군, 운동과 hesperetin 복합처치군으로 구분하여 운동은 점증부하원리에 의해 최종적으로 27m/min으로 30분간 주당 5회 실시하였으며, hesperetin은 식이내 0.02% 첨가하였다. 각 집단의 체중 증가는 통계적으로 유의한 차이를 보이지 않았으나, 장간막, 복강내 지방 및 총지방량은 운동에 의해 유의하게 감소되었다. 혈장 총콜레스테롤, LDL-C 농도 및 동맥경화지수는 hesperetin의 처치로 유의하게 감소되었으며, 중성지방 농도는 운동에 의해 유의하게 감소된 것으로 나타났다. 본 연구에서 혈중지질에 대한 운동과 hesperetin의 상호관계는 나타나지 않았으나 각각 독립적인 혈중지질개선 효과를 보여주었으므로 운동과 hesperetin의 복합처치는 혈중지질개선에 대한 상호보완적인 효과를 기대할 수 있을 것으로 사료된다. Hesperetin, a kind of citrus flavonoids, have been identified that exerts plasma lipid lowering effect in rodents. We examined the combined effects of treadmill exercise and the 0.02% hesperetin supplementation for 6 weeks on plasma lipid profile in SD rats which fed a high cholesterol diet. Incremental treadmill running was performed at 27 m·min-1, a half hour·day-1, and five days·week-1 as a final intensity. Plasma total cholesterol, low density lipoprotein cholesterol, and atherogenic index level lowered with hesperetin treatment and plasma triacylglycerol lowered with exercise treatment(p<.05). There is no interaction between the hesperetin and the exercise treatment. This results suggested that the combined treatment of treadmill exercise and hesperetin supplementation might give a beneficial effects on plasma lipid profile complementarily.

마우스 대식세포 RAW 264.7 세포주에서 hesperetin에 의한 p38 MAPK와 ERK1/2를 통한 염증반응 조절

이승훈(Seung-Hoon Lee),이은주(Eun-Joo Lee),정정욱(Chungwook Chung),손호용(Ho-Yong Sohn),김종식(Jong-Sik Kim) 한국생명과학회 2019 생명과학회지 Vol.29 No.1

이전 연구에서 전통주 주박 ethyl acetate 분획물로부터 11개의 순수물질을 분리 동정하였다. 11개의 순수물질은 caffeic acid, coumaric acid, D-mannitol, ferulic acid, hesperetin, hesperidin, naringenin, naringin, sinapic acid, syringic acid, 그리고 vanilic acid로 동정되었다. 이번 연구에서는 그들의 항염증 활성을 연구하기 위하여 LPS로 활성화된 RAW 264.7 세포에서 nitric oxide (NO) 생산을 측정하였다. 11개의 순수물질 중 hesperetin과 naringenin이 가장 높은 NO 생성 억제를 보여주었다. 또한, hesperetin은 세포 생존율에 영향 없이 농도의존적으로 NO 생산을 저해하였다. 그리고, hesperetin은 농도의존적으로 염증유전자인 iNOS의 발현을 농도의존적으로 억제한 반면, COX-2 단백질의 발현에는 영향을 주지 않았다. 게다가, hesperetin은 p38 MAPK와 ERK1/2의 인산화를 억제한 반면 JNK의 인산화에는 영향을 주지 못했다. 이러한 결과는 hesperetin은 항염증 활성을 가지며, 이러한 항염증 활성은 p38 MAPK와 ERK1/2 경로를 억제함으로써 일어난다는 것을 나타낸다. In a previous study, we isolated 11 different kinds of compounds from ethyl acetate fractions of lees (jubak) which is a by-product of Korean traditional wine production. These compounds were identified as caffeic acid, coumaric acid, D-mannitol, ferulic acid, hesperetin, hesperidin, naringenin, naringin, sinapic acid, syringic acid, and vanilic acid. To evaluate their anti-inflammatory activities in an in vitro model, nitric oxide (NO) production was measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells after the treatment of these cells with each compound. Among the various chemicals, hesperetin and naringenin showed the highest inhibition of NO production in the LPS-activated RAW 264.7 cells. Hesperetin was chosen for further study because of its strong anti-inflammatory activity and because the mechanisms underlying its anti-inflammatory properties still remain unclear. Our results showed that hesperetin dramatically inhibited NO production in a dose-dependent manner as compared with in an LPS-only treated group, without affecting cell viability. In addition, hesperetin reduced the protein expression of the pro-inflammatory gene inducible nitric oxide synthase (iNOS) in a dose-dependent manner, whereas it did not affect cyclooxygenase-2 (COX-2) expression. Furthermore, hesperetin inhibited phosphorylation of p38 mitogen- activated protein kinase (MAPK) and extracellular signal regulated kinase (ERK) 1/2, whereas it did not affect phosphorylation of c-jun N- terminal kinase (JNK). The results indicated that hesperetin regulated the LPS-induced inflammatory response by suppressing p38 MAPK and ERK1/2 signaling. Overall, our results may help to understand the mechanisms underlying the anti-inflammatory activity mediated by hesperetin.

Hesperidin과 Hesperetin의 간 손상 동물모델에서 산화적 스트레스에 대한 간 보호 효과

김지현(Ji Hyun Kim),이여(Li Li),김미숙(Mi Suk Kim),조은주(Eun Ju Cho),김현영(Hyun Young Kim),최진상(Jine Shang Choi) 한방비만학회 2022 한방비만학회지 Vol.22 No.1

Objectives: To investigate the protective effect of hesperidin and hesperetin against oxidative stress in 2,2'-azobis (2-aminopropane) dihydrochloride (AAPH)-induced liver toxicity in rats. Methods: Hesperidin or hesperetin (200 mg/kg/day, respectively) was orally administered for 7 days once daily in rats. Subsequently, AAPH (50 mg/kg/day) was administered intraperitoneally. Lipid peroxidation, nitric oxide production, catalase activity, and protein expressions of nuclear factor-kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) in the liver tissues were measured. Results: Administration of hesperidin and hesperetin significantly decreased serum aspartate transaminase and alanine transaminase levels in AAPH-induced oxidative stress liver tissues compared with control group. Lipid peroxidation and nitric oxide (NO) production were also significantly reduced by hesperidin and hesperetin in AAPH-induced oxidative stress liver tissues. In particular, lipid peroxidation levels of hesperetin-administered group significantly decreased to 5.02 nmole/mg protein in oxidative stress rats. Hesperidin and hesperetin significantly increased antioxidant activity, such as that of catalase. Furthermore, administration of hesperidin and hesperetin substantially down-regulated the expression of NF-κB and iNOS in liver tissues. Administration of hesperidin reduced NO levels and iNOS expression more than in the hesperetin-administered group. Conclusions: Administration of hesperidin and hesperetin led to a reduction in AAPH-induced liver toxicity by regulating oxidative stress.

Hesperetin suppresses LPS/high glucose-induced inflammatory responses via TLR/MyD88/NF-κB signaling pathways in THP-1 cells

Aeri Lee,HyunJi Gu,Min-Hee Gwon,Jung-Mi Yun 한국영양학회 2021 Nutrition Research and Practice Vol.15 No.5

BACKGROUND/OBJECTIVES: Unregulated inflammatory responses caused by hyperglycemia may induce diabetes complications. Hesperetin, a bioflavonoid, is a glycoside in citrus fruits and is known to have antioxidant and anticarcinogenic properties. However, the effect of inflammation on the diabetic environment has not been reported to date. In this study, we investigated the effect of hesperetin on proinflammatory cytokine secretion and its underlying mechanistic regulation in THP-1 macrophages with co-treatment LPS and hyperglycemic conditions. MATERIALS/METHODS: THP-1 cells differentiated by PMA (1 μM) were cultured for 48 h in the presence or absence of hesperetin under normoglycemic (5.5 mM/L glucose) or hyperglycemic (25 mM/L glucose) conditions and then treated with LPS (100 ng/mL) for 6 h before harvesting. Inflammation-related proteins and mRNA levels were evaluated by enzyme-linked immunosorbent assay, western blot, and quantitative polymerase chain reaction analyses. RESULTS: Hesperetin (0–100 μM, 48 h) treatment did not affect cell viability. The tumor necrosis factor-α and interleukin-6 levels increased in cells co-treated with LPS under hyperglycemic conditions compared to normoglycemic conditions, and these increases were decreased by hesperetin treatment. The TLR2/4 and MyD88 activity levels increased in cells co-treated with LPS under hyperglycemic conditions compared to normoglycemic conditions; however, hesperetin treatment inhibited the TLR2/4 and MyD88 activity increases. In addition, nuclear factor-κB (NF-κB) and Acetyl-NF-κB levels increased in response to treatment with LPS under hyperglycemic conditions compared to normoglycemic conditions, but those levels were decreased when treated with hesperetin. SIRT3 and SIRT6 expressions were increased by hesperetin treatment. CONCLUSIONS: Our results suggest that hesperetin may be a potential agent for suppressing inflammation in diabetes.

Neuroprotective effects of hesperetin on H2O2-induced damage in neuroblastoma SH-SY5Y cells

Moon Ha-Rin,Yun Jung-Mi 한국영양학회 2023 Nutrition Research and Practice Vol.17 No.5

BACKGROUND/OBJECTIVES: Oxidative stress is a fundamental neurodegenerative disease trigger that damages and decimates ner ve cells. Neurodegenerative diseases are chronic central ner vous system disorders that progress and result from neuronal degradation and loss. Recent studies have extensively focused on neurodegenerative disease treatment and prevention using dietar y compounds. Heseperetin is an aglycone hesperidin form with various physiological activities, such as anti-inflammation, antioxidant, and antitumor. However, few studies have considered hesperetin’s neuroprotective effects and mechanisms; thus, our study investigated this in hydrogen peroxide (H2O2)-treated SH-SY5Y cells. MATERIALS/METHODS: SH-SY5Y cells were treated with H2O2 (400 µM) in hesperetin absence or presence (10–40 µM) for 24 h. Three-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays detected cell viability, and 4′,6-diamidino-2-phenylindole staining allowed us to obser ve nuclear morphology changes such as chromatin condensation and apoptotic nuclei. Reactive oxygen species (ROS) detection assays measured intracellular ROS production; Griess reaction assays assessed nitric oxide (NO) production. Western blotting and quantitative polymerase chain reactions quantified corresponding mRNA and proteins. RESULTS: Subsequent experiments utilized various non-toxic hesperetin concentrations, establishing that hesperetin notably decreased intracellular ROS and NO production in H2O2-treated SH-SY5Y cells (P < 0.05). Furthermore, hesperetin inhibited H2O2-induced inflammation-related gene expression, including interluekin-6, tumor necrosis factor-α, and nuclear factor kappa B (NF-κB) p65 activation. In addition, hesperetin inhibited NF- κB translocation into H2O2-treated SH-SY5Y cell nuclei and suppressed mitogen-activated protein kinase protein expression, an essential apoptotic cell death regulator. Various apoptosis hallmarks, including shrinkage and nuclear condensation in H2O2-treated cells, were suppressed dose-dependently. Additionally, hesperetin treatment down-regulated Bax/ Bcl-2 expression ratios and activated AMP-activated protein kinase-mammalian target of rapamycin autophag y pathways. CONCLUSION: These results substantiate that hesperetin activates autophagy and inhibits apoptosis and inflammation. Hesperetin is a potentially potent dietar y agent that reduces neurodegenerative disease onset, progression, and prevention.

Hesperidin과 Hesperetin의 AAPH로 산화적 손상을 유도한 동물 모델에서 항산화 활성 조절을 통한 신장 보호 효과

김지현,이여,김미숙,조은주,김현영 경상국립대학교 농업생명과학연구원 2022 농업생명과학연구 Vol.56 No.1

본 연구는 hesperidin과 hesperidin의 aglycone 형태인 hesperetin의 2,2'-azobis (2-aminopropane) dihydrochloride (AAPH)에 의해 신장독성이 유도된 쥐에서 신장 보호 효과에 대해 연구하였다. Hesperidin과 hesperetin은 200 mg/kg/day의 농도로 7일간 위내투여하였으며, AAPH를복강주사하여 급성 신장 손상을 유도하였다. 이 후 쥐의 신장 조직에서 지질과산화 함량, nitric oxide (NO) 생성량, catalase 효소 활성을 측정하였으며, nuclear factor-kappa B (NF-κB) 및 inducible nitric oxide synthase (iNOS) 단백질 발현량을 측정하였다. AAPH로 신장 독성을 유도한control군의 신장 내 지질과산화 및 NO 생성량은 신장 독성을 유도하지 않은 normal군에 비해 유의적으로 증가하여 산화적 손상이 유도됨을확인하였다. 반면 hesperidin과 hesperetin를 투여했을 때 신장 내 지질과산화 및 NO 생성량이 control군에 비해 유의적으로 감소하여 산화적스트레스 개선 효과를 확인하였다. Hesperidin과 hesperetin을 투여한 군의 경우 신장 내 항산화 효소인 catalase 활성을 유의적으로 증가시켰다. 뿐만 아니라, hesperidin과 hesperetin의 투여는 AAPH로 신장독성을 유도한 control군 100% 대비 NF-κB 단백질을 각각 66% 및 71%로, iNOS 단백질 발현을 각각 46% 및 33%로 억제시켰다. 따라서 본 연구를 통해 hesperidin과 hesperetin의 투여가 항산화 활성 조절을 통해 AAPH로유도된 신장 독성을 억제하는 것을 알 수 있었다.

Antioxidant and Neuroprotective Effects of Hesperidin and its Aglycone Hesperetin

Cho, Jung-Sook The Pharmaceutical Society of Korea 2006 Archives of Pharmacal Research Vol.29 No.8

The present study evaluated antioxidant and neuroprotective activities of hesperidin, a flavanone mainly isolated from citrus fruits, and its aglycone hesperetin using cell-free bioassay system and primary cultured rat cortical cells. Both hesperidin and hesperetin exhibited similar patterns of 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities. While hesperidin was inactive, hesperetin was found to be a potent antioxidant, inhibiting lipid peroxidation initiated in rat brain homogenates by $Fe^{2+}$ and L-ascorbic acid. In consistence with these findings, hesperetin protected primary cultured cortical cells against the oxidative neuronal damage induced by $H_2O_2$ or xanthine and xanthine oxidase. In addition, it was shown to attenuate the excitotoxic neuronal damage induced by excess glutamate in the cortical cultures. When the excitotoxicity was induced by the glutamate receptor subtype-selective ligands, only the N-methyl-D-aspartic acid-induced toxicity was selectively and markedly inhibited by hesperetin. Furthermore, hesperetin protected cultured cells against the $A_{{\beta}(25-35)}-induced$ neuronal damage. Hesperidin, however, exerted minimal or no protective effects on the neuronal damage tested in this study. Taken together, these results demonstrate potent antioxidant and neuroprotective effects of hesperetin, implying its potential role in protecting neurons against various types of insults associated with many neurodegenerative diseases.

Antioxidant and Neuroprotective Effects of Hesperidin and its Aglycone Hesperetin

조정숙 대한약학회 2006 Archives of Pharmacal Research Vol.29 No.8

The present study evaluated antioxidant and neuroprotective activities of hesperidin, a flavanone mainly isolated from citrus fruits, and its aglycone hesperetin using cell-free bioassay system and primary cultured rat cortical cells. Both hesperidin and hesperetin exhibited similar patterns of 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities. While hesperidin was inactive, hesperetin was found to be a potent antioxidant, inhibiting lipid peroxidation initiated in rat brain homogenates by Fe2+ and L-ascorbic acid. In consistence with these findings, hesperetin protected primary cultured cortical cells against the oxidative neuronal damage induced by H2O2 or xanthine and xanthine oxidase. In addition, it was shown to attenuate the excitotoxic neuronal damage induced by excess glutamate in the cortical cultures. When the excitotoxicity was induced by the glutamate receptor subtype-selective ligands, only the N-methyl-Daspartic acid-induced toxicity was selectively and markedly inhibited by hesperetin. Furthermore, hesperetin protected cultured cells against the Aβ(25-35)-induced neuronal damage. Hesperidin, however, exerted minimal or no protective effects on the neuronal damage tested in this study. Taken together, these results demonstrate potent antioxidant and neuroprotective effects of hesperetin, implying its potential role in protecting neurons against various types of insults associated with many neurodegenerative diseases.

Hesperidin과 Hesperetin의 in vitro에서의 항산화 효과

김현영,이여,조은주 대한암예방학회 2010 Journal of cancer prevention Vol.15 No.4

In this study, we investigated the antioxidant activity of hesperidin and hesperetin, which are the active compounds from Citrus junos, under in vitro. We measured the scavenging activities of hesperidin and hesperetin on 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical (․OH), nitric oxide (NO) and the inhibition activities on oxidation of protein and lipid under in vitro. Hesperidin and hesperetin dose-dependently scavenged DPPH, ․OH and nitric oxide (NO). In addition, hesperidin and hesperetin significantly reduced oxidation of protein and lipid induced by 2,2'-azobis(2-aminopropane) dihydrochloride. Hesperidin had stronger activity in protein oxidation, on the other hand hesperetin had stronger activity in lipid oxidation. The present study indicates that hesperidin and hesperetin would have antioxidant activities by protecting against oxidative damage induced by free radicals. (Cancer Prev Res 15, 333-339, 2010)

Molecular Mechanism Underlying Hesperetin-induced Apoptosis by in silico Analysis and in Prostate Cancer PC-3 Cells

Sambantham, Shanmugam,Radha, Mahendran,Paramasivam, Arumugam,Anandan, Balakrishnan,Malathi, Ragunathan,Chandra, Samuel Rajkumar,Jayaraman, Gopalswamy Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.7

Aim: To investigate the molecular mechanisms underlying triggering of apoptosis by hesperetin using in silico and in vitro methods. Methods: The mechanism of binding of hesperetin with NF-${\kappa}B$ and other apoptotic proteins like BAX, BAD, $BCL_2$ and $BCL_{XL}$ was analysed in silico using Schrodinger suite 2009. In vitro studies were also carried out to evaluate the potency of hesperetin in inducing apoptosis using the human prostate cancer PC-3 cell line. Results: Hesperetin was found to exhibit high-affinity binding resulting from greater intermolecular forces between the ligand and its receptor NF-${\kappa}B$ (-7.48 Glide score). In vitro analysis using MTT assay confirmed that hesperetin reduced cell proliferation ($IC_{50}$ values of 90 and $40{\mu}M$ at 24 and 48h respectively) in PC-3 cells. Hesperetin also downregulated expression of the anti-apoptotic gene $BCL_{XL}$ at both mRNA and protein levels and increased the expression of pro-apoptotic genes like BAD at mRNA level and BAX at mRNA as well as protein levels. Conclusion: The results suggest that hesperetin can induce apoptosis by inhibiting NF-${\kappa}B$.