Hesperidin structurally modified by gamma irradiation induces apoptosis in murine melanoma B16BL6 cells and inhibits both subcutaneous tumor growth and metastasis in C57BL/6 mice

Byun, Eui-Baek,Kim, Hye-Min,Song, Ha-Yeon,Kim, Woo Sik Elsevier 2019 Food and chemical toxicology Vol.127 No.-

<P><B>Abstract</B></P> <P>Hesperidin is a flavonoid which occurs in citrus fruits. Hesperidin was gamma-irradiated at doses of 0, 30, 70, and 150 kGy. Gamma irradiation induced a decreased hesperidin peak, and a new radiolytic peak that gradually increased up to 150 kGy. The new radiolytic peak was fractionated, and the fractionated hesperidin derivative was used for subsequent experiments. Hesperidin gamma-irradiated at 150 kGy was toxic toward B16BL6 cells, but not toward bone marrow-derived macrophages. This cytotoxicity was exerted via induction of apoptosis, as reflected by the high population of double-positive cells, increased sub-G1 phase cells, depolarization of matrix metalloproteinase, production of reactive oxygen species, weakness of cell adhesion, changes in cell morphology, and inhibition of B16BL6 cell migration. Furthermore, 150 kGy gamma-irradiated hesperidin decreased the expression of Bcl-2 and pro-caspases-3 and -9, increased the expression of Bax and cytosolic cytochrome <I>c</I>, and increased the cleavage of poly ADP ribose polymerase. <I>In vitro</I> mechanistic study revealed that 150 kGy gamma-irradiated hesperidin achieved significantly greater inhibition of lung metastasis and growth of melanoma B16BL6 cells in C57BL/6 mice than non-irradiated intact hesperidin did. These results suggest that the structural modification of hesperidin induced by gamma irradiation could facilitate the development of anti-cancer drugs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Gamma irradiation induced structural transformation of hesperidin. </LI> <LI> Gamma-irradiated hesperidin effectively induced apoptosis than unirradiated-hesperidin in B16BL6 melanoma cells through mitochondrial pathway. </LI> <LI> Gamma-irradiated hesperidin exhibited preventive and therapeutic effects in lung metastasis and tumor growth mice model. </LI> <LI> Gamma-irradiated hesperidin might be a candidate for cancer therapy. </LI> </UL> </P>

Enhanced Bioavailability of Verapamil after Oral Administration with Hesperidin in Rats

Yong-Ji Piao,Jun-Shik Choi 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.4

The aim of this study was to investigate the effects of hesperidin on the pharmacokinetics of verapamil and its major metabolite, norverapamil, in rats. The pharmacokinetic parameters of verapamil and norverapamil in rats were measured after the oral administration of verapamil (9 mg/kg) in the presence or absence of hesperidin (3 or 10 mg/kg). Compared to the control group, the presence of hesperidin significantly (p<0.01) increased the area under the plasma concentration-time curve (AUC) of verapamil by 71.1–96.8% and the peak concentration (Cmax) of verapamil by 98.3-105.2%. Hesperidin significantly (p<0.01) decreased the total plasma clearance (CL/F) of verapamil by 41.6-49.2% in rats. However there was no significant change in the time to reach the peak plasma concentration (Tmax), the elimination rate constant (Kel) and the terminal half-life (T1/2) of verapamil in the presence of hesperidin. The AUC and Cmax of norverapamil were significantly (p<0.05) higher in rats coadministrated with hesperidin than those of the control. Consequently hesperidin significantly enhanced bioavailability of verapamil in rats. These results might be due to the decreased efflux and metabolism of verapamil in the intestine. Drug interactions should be concerned in the clinical setting when verapamil is used concomitantly with hesperidin or hesperidin-containing dietary

Hesperidin depolarizes the pacemaker potentials through 5-HT4 receptor in murine small intestinal interstitial cells of Cajal

황민우,김정남,김병주 한국통합생물학회 2020 Animal cells and systems Vol.24 No.2

Hesperidin, a citrus flavonoid, can exert numerous beneficial effects on human health. Interstitial cells of Cajal (ICC) are pacemaker cells in the gastrointestinal (GI) tract. In the present study, we investigated potential effects of hesperidin on pacemaker potential of ICC in murine small intestine and GI motility. A whole-cell patch-clamp configuration was used to record pacemaker potential in ICC, and GI motility was investigated in vivo by recording gastric emptying (GE) and intestinal transit rate (ITR). Hesperidin depolarized pacemaker potentials of ICC in a dosedependent manner. Pre-treatment with methoctramine or 4-DAMP did not inhibit hesperidininduced pacemaker potential depolarization. Neither a 5-HT3 receptor antagonist (Y25130) nor a 5-HT7 receptor antagonist (SB269970) reduced the effect of hesperidin on ICC pacemaker potential, whereas the 5-HT4 receptor antagonist RS39604 was found to inhibit this effect. In the presence of GDP–β–S, hesperidin-induced pacemaker potential depolarization was inhibited. Moreover, in the presence of U73122 and calphostin C, hesperidin did not depolarize pacemaker potentials. Furthermore, hesperidin accelerated GE and ITR in vivo. These results imply that hesperidin depolarized ICC pacemaker potential via 5-HT4 receptors, G protein, and PLC/PKC dependent pathways and that it increased GI motility. Therefore, hesperidin may be a promising novel drug to regulate GI motility.

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.

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.

Enhanced Bioavailability of Verapamil after Oral Administration with Hesperidin in Rats

Piao, Yong-Ji,Choi, Jun-Shik 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.4

The aim of this study was to investigate the effects of hesperidin on the pharmacokinetics of verapamil and its major metabolite, norverapamil, in rats. The pharmacokinetic parameters of verapamil and norverapamil in rats were measured after the oral administration of verapamil (9 mg/kg) in the presence or absence of hesperidin (3 or 10 mg/kg). Compared to the control group, the presence of hesperidin significantly (p<0.01) increased the area under the plasma concentration-time curve (AUC) of verapamil by 71.1-96.8% and the peak concentration $(C_{max})$ of verapamil by 98.3-105.2%. Hesperidin significantly (p<0.01) decreased the total plasma clearance (CL/F) of verapamil by 41.6-49.2% in rats. However there was no significant change in the time to reach the peak plasma concentration $(T_{max})$, the elimination rate constant $(K_{el})$ and the terminal half-life $(T_{1/2})$ of verapamil in the presence of hesperidin. The AUC and $C_{max}$ of norverapamil were significantly (p<0.05) higher in rats coadministrated with hesperidin than those of the control. Consequently hesperidin significantly enhanced bioavailability of verapamil in rats. These results might be due to the decreased efflux and metabolism of verapamil in the intestine. Drug interactions should be concerned in the clinical setting when verapamil is used concomitantly with hesperidin or hesperidin-containing dietary.

Hesperidin Induces Apoptosis in SNU-668, Human Gastric Cancer Cells

Park, Hae-Jeong,Ra, Je-Hyun,Han, Mi-Young,Chung, Joo-Ho The Korean Society of Toxicogenomics and Toxicopro 2007 Molecular & cellular toxicology Vol.3 No.1

Hesperidin, known as a flavonoid constituent of citrus, has been known to reduce the proliferation of several cancer cells. We investigated whether hesperidin-induced cell death on SNU-668, human gastric cancer cells. The cytotoxicity of hesperidin on SNU-668 cells was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at the concentration of 1, 10, 50, and 100 ${\mu}M$. Cell viability by hesperidin was 53.18$\pm$2.85% of control value at 100 ${\mu}M$. The cell death by hesperidin showed apoptotic features, which were confirmed using a combination of 4, 6-diamidino-2-phenylindole (DAPI) staining and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. In the apoptosis-regulating genes, treatment of hesperidin decreased mRNA expression of B-cell CLL/lymphoma 2 (BCL2), whereas expression of BCL2-associated X protein (BAX) was increased. The mRNA expression and the activity of caspase3 (CASP3), a major apoptotic factor, was significantly increased by hesperidin treatment. These results suggest that hesperidin could induce apoptosis through CASP3 activation on SNU-668, human gastric cancer cells.

Effect of Hesperidin Supplementation on Lipid and Antioxidant Metabolism in Ethanol-fed Rats

Soon-Ja Kim,Hyun-Ju Seo,Hye-Jin Kim,Yun-Young Cho,Eun-Young Kwon,Hyosun Lee,Myung-Sook Choi 한국식품영양과학회 2006 Preventive Nutrition and Food Science Vol.11 No.4

This study examined the effect of hesperidin supplementation with an ethanol diet on lipid and antioxidant metabolism in rats. Male Sprague-Dawley rats were divided into two groups (n=10), and were assigned to one of two dietary categories: E?, ethanol diet (50 g/L) for 8 wks; E?H₄, ethanol diet for the first 4 wks and hesperidin (0.02%, w/w) supplemented ethanol diet for the last 4 wks. The plasma and hepatic lipids, hepatic cholesterol regulating enzyme activity, hepatic antioxidant enzyme activity and lipid peroxidation were determined. Supplementation with hesperidin for the last 4 wks during the 8 wks period of the ethanol diet, significantly increased the ADH activity. In conjunction with the chronic administration of ethanol, hesperidin supplementation resulted in a significant decrease in the hepatic cholesterol and triglyceride concentrations compared to the E? group. The hepatic HMG-CoA reductase and ACAT activities were significantly lower in the hesperidin-supplemented group. When comparing hepatic antioxidant enzyme activities, SOD, GSH-Px, and G6PD activities and GSH level were significantly higher in the E?H₄ group than in the E? group. Plasma TBARS levels were significantly lower in rats fed ethanol with hesperidin compared to the rats fed only ethanol; however, the hepatic TBARS levels were not significantly different between the groups. Accordingly, the additional hesperidin supplement with an ethanol diet might be effective for improving the hepatic lipid metabolism and antioxidant defense system.

Effect of Hesperidin Supplementation on Lipid and Antioxidant Metabolism in Ethanol-fed Rats

Kim, Soon-Ja,Seo, Hyun-Ju,Kim, Hye-Jin,Cho, Yun-Young,Kwon, Eun-Young,Lee, Hyo-Sun,Choi, Myung-Sook The Korean Society of Food Science and Nutrition 2006 Preventive Nutrition and Food Science Vol.11 No.4

This study examined the effect of hesperidin supplementation with an ethanol diet on lipid and antioxidant metabolism in rats. Male Sprague-Dawley rats were divided into two groups (n=10), and were assigned to one of two dietary categories: $E_8$, ethanol diet (50 g/L) for 8 wks; $E_8H_4$, ethanol diet for the first 4 wks and hesperidin (0.02%, w/w) supplemented ethanol diet for the last 4 wks. The plasma and hepatic lipids, hepatic cholesterol regulating enzyme activity, hepatic antioxidant enzyme activity and lipid peroxidation were determined. Supplementation with hesperidin for the last 4 wks during the 8 wks period of the ethanol diet, significantly increased the ADH activity. In conjunction with the chronic administration of ethanol, hesperidin supplementation resulted in a significant decrease in the hepatic cholesterol and triglyceride concentrations compared to the $E_8$ group. The hepatic HMG-CoA reductase and ACAT activities were significantly lower in the hesperidin-supplemented group. When comparing hepatic antioxidant enzyme activities, SOD, GSH-Px, and G6PD activities and GSH level were significantly higher in the $E_8H_4$ group than in the E8 group. Plasma TBARS levels were significantly lower in rats fed ethanol with hesperidin compared to the rats fed only ethanol; however, the hepatic TBARS levels were not significantly different between the groups. Accordingly, the additional hesperidin supplement with an ethanol diet might be effective for improving the hepatic lipid metabolism and antioxidant defense system.

Anti-inflammatory Effects and its Mechanisms of Hesperidin in an Asthmatic Mouse Model Induced by Ovalbumin

Jeong Hyun Chang 대한의생명과학회 2010 Biomedical Science Letters Vol.16 No.2

Hesperidin, a member of the flavanone group of flavonoids, can be isolated in large amounts from the rinds of some citrus species [e.g., Citrus aurantium L. (bitter orange), Citrus sinensis L. (sweet orange) and Citrus unshiu Marcov. (satsuma mandarin)], and has been reported to have anticarcinogenic, antihypotensive and antimicrobial properties. Despite the efficacy of these polyphenolic compounds as immune modulators, the effects of the flavonoids are poorly understood about allergic effect. In this study, we investigated whether hesperidin could influence on Th1 and Th2 balance. Allergic reactions included an increase in the number of eosinophils in bronchoalveolar lavage (BAL) fluid, an increase in inflammatory cell infiltration into the lung tissue around blood vessels and airways, airway luminal narrowing, the development of airway hyper-responsiveness (AHR). The administration of hesperidin before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions. Accordingly, this study may provide evidence that hesperidin plays a critical role in the amelioration of the pathogenetic process of asthma in mice. These findings provide new insight into the immunopharmacological role of hesperidin in terms of its effects in a murine model of asthma, and also broaden current perspectives in our understanding of the immunopharmacological functions of hesperidin.