Hawthorn ethanolic extracts with triterpenoids and flavonoids exert hepatoprotective effects and suppress the hypercholesterolemia-induced oxidative stress in rats

Document Type: Original Article


1 Department of Histology and Embryology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

3 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Urmia Medical University of Sciences, Urmia, Iran

4 Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

5 Department of Pathology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

6 Division of Veterinary Pharmacy, Pharmacology and Toxicology, Faculty of Science, Utrecht University, The Netherlands

7 Division of Pharmacology, Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, The Netherlands


Objective(s):The current study was aimed to determine the bioactive constituents and biological effects of the Crataegus monogyna ethanolic extracts from bark, leaves and berries on hypercholesterolemia.
Materials and Methods: Oleanolic acid, ursolic acid, quercetin and lupeol concentrations were quantified by HPLC. Total phenol content and radical scavenging activity of extracts were also measured. The hypocholesterolemic, antioxidant, and hepatoprotective effects of the extracts were examined in hypercholesterolemic rats and compared with orlistat.
Results: The highest phenol content, oleanolic acid, quercetin and lupeol levels and free radical scavenging potency were found in the bark extract, and the highest ursolic acid level was found in the berries extract. Orlistat and extracts significantly (P<0.05) lowered the hypercholesterolemia-increased serum level of hepatic enzymes and lipid peroxidation level. Hawthorn’s extracts protected from hepatic thiol depletion and improved the lipid profile and hepatic damages.
Conclusion: Data suggested that hawthorn’s extracts are able to protect from hypercholesterolemia-induced oxidative stress and hepatic injuries. Moreover, the hypocholesterolemic effect of extracts was found comparable to orlistat.


1. Petkov V. Plants with hypotensive, antiathero-matous and coronarodilatating action. Am J Chin Med 1979; 7:197-236.

2. Rigelsky JM, Sweet BV. Hawthorn: pharmacology and therapeutic uses. Am J Health Syst Pharm 2002; 59:417-422.

3. Tassell MC, Kingston R, Gilroy D, Lehane M, Furey A. Hawthorn (Crataegus spp.) in the treatment of cardiovascular disease. Pharmacogn Rev 2010; 4:32.

4. Rajendran S, Deepalakshmi P, Parasakthy K, Devaraj H, Devaraj SN. Effect of tincture of Crataegus on the LDL-receptor activity of hepatic plasma membrane of rats fed an atherogenic diet. Atherosclerosis 1996; 123:235-241.

5. Lin Y, Vermeer MA, Trautwein EA. Triterpenic acids present in hawthorn lower plasma cholesterol by inhibiting intestinal ACAT activity in hamsters. Evid Based Complement Alternat Med 2011: article ID 801272; doi: 10.1093/ecam/nep007

6. Urbonavičiūtė A, Jakštas V, Kornyšova O, Janulis V, Maruška A. Capillary electrophoretic analysis of flavonoids in single-styled hawthorn (Crataegus monogyna Jacq.) ethanolic extracts. J Chromatogr A 2006; 1112:339-344.

7. Zhang Z, Chang Q, Zhu M, Huang Y, Ho WK, Chen Z-Y. Characterization of antioxidants present in hawthorn fruits. J Nutr Biochem 2001; 12:144-152.

8. Liu P, Yang B, Kallio H. Characterization of phenolic compounds in Chinese hawthorn (Crataegus pinnatifida Bge. var. major) fruit by high performance liquid chromatography–electrospray ionization mass spectro-metry. Food Chem 2010; 121:1188-1197.

9. D'Mello PM, Joshi UJ, Shetgiri PP, Dasgupta TK, Darji KK. A simple HPLC method for quantitation of quercetin in herbal extracts. J AOAC Int 2011; 94:100-105.

10. Saboo S, Taapadiya G, Khadabadi S. Antioxidant and hepatoprotective potential of isolated fraction of Rivea hypocrateriformis. J Biol Sci 2011; 11:428-432.

11. Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzymol 1999; 299:152-178.

12. Mensor LL, Menezes FS, Leitão GG, Reis AS, Santos TCd, Coube CS, et al. Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method. Phytother Res 2001; 15:127-130.

13. Lu LS, Wu CC, Hung LM, Chiang MT, Lin CT, Lin CW,  et al. Apocynin alleviated hepatic oxidative burden and reduced liver injury in hypercholesterolaemia. Liver Int 2007; 27:529-537.

14. Hosseinimehr SJ, Azadbakht M, Abadi AJ. Protective effect of hawthorn extract against genotoxicity induced by cyclophosphamide in mouse bone marrow cells. Environ Toxicol Pharmacol 2008; 25:51-56.

15. Caner M, Dogruman H, Taskin E, Kandil A, Demirci C. Effects of orlistat and its relationship with nitric oxide in the small intestinal mucosa. Chin J Physiol 2005; 48:217-222.

16. Ranjbar A, Khorami S, Safarabadi M, Shahmoradi A, Malekirad AA, Vakilian K, et al. Antioxidant activity of Iranian Echium amoenum Fisch & CA Mey flower decoction in humans: a cross-sectional before/after clinical trial. Evid Based Complement Alternat Med 2006; 3:469-473.

17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193:265-275.

18. Niehaus W, Samuelsson B. Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. Eur J Biochem 1968; 6:126-130.

19. Wang H, Zhang Z, Guo Y, Sun P, Lv X, Zuo Y. Hawthorn fruit increases the antioxidant capacity and reduces lipid peroxidation in senescence-accelerated mice. Eur Food Res Technol 2011; 232:743-751.

20. Kwok CY, Wong CNY, Yau MYC, Yu PHF, Au ALS, Poon CCW, et al. Consumption of dried fruit of Crataegus pinnatifida (hawthorn) suppresses high-cholesterol diet-induced hypercholesterolemia in rats. J Funct Foods 2010; 2:179-186.

21. Kushner R. Medical management of obesity. Semin Gastrointest Dis 2002; 13:123-132

22. Silva JRdA, Rezende CM, Pinto ÂC, Pinheiro ML, Cordeiro MC, Tamborini E, et al. Triterpenic esters from himatanthus sucuuba (Spruce) Woodson. Quim Nova 1998; 21:702-704.

23. Baskar R, Meenalakshmi Malini M, Varalakshmi P, Balakrishna K, Bhima Rao R. Effect of lupeol isolated from Crataeva nurvala stem bark against free radical-induced toxicity in experimental urolithiasis. Fitoterapia 1996; 67:121-125.

24. Saratha V, Subramanian SP. Lupeol, a triterpenoid isolated from Calotropis gigantea latex ameliorates
the primary and secondary complications of FCA induced adjuvant disease in experimental rats. Inflammopharmacology 2012; 20:27-37.

25. Nagaraj M, Sunitha S, Varalakshmi P. Effect of lupeol, a pentacyclic triterpene, on the lipid peroxidation and antioxidant status in rat kidney after chronic cadmium exposure. J Appl Toxicol 2000; 20:413-417.

26. Saleem M, Afaq F, Adhami VM, Mukhtar H. Lupeol modulates NF- B and PI3K/Akt pathways and inhibits skin cancer in CD-1 mice. Oncogene 2004; 23:5203-5214.

27. Saleem M, Kaur S, Kweon M-H, Adhami VM, Afaq F, Mukhtar H. Lupeol, a fruit and vegetable based triterpene, induces apoptotic death of human pancreatic adenocarcinoma cells via inhibition of Ras signaling pathway. Carcinogenesis 2005; 26:1956-1964.

28. Sudhahar V, Kumar SA, Varalakshmi P. Role of lupeol and lupeol linoleate on lipemic–oxidative stress in experimental hypercholesterolemia. Life Sci 2006; 78:1329-1335.

29. Itoh M, Hiwatashi K, Abe Y, Kimura F, Toshima G, Takahashi J, et al. Lupeol reduces triglyceride and cholesterol synthesis in human hepatoma cells. Phytochem Lett 2009; 2:176-178.

30. Radhiga T, Rajamanickam C, Senthil S, Pugalendi KV. Effect of ursolic acid on cardiac marker enzymes, lipid profile and macroscopic enzyme mapping assay in isoproterenol-induced myocardial ischemic rats. Food Chem Toxicol 2012; 50:3971-3977.

31. Azevedo MF, Camsari Ç, Sá CM, Lima CF, Fernandes Ferreira M, Pereira Wilson C. Ursolic acid and luteolin 7 glucoside improve lipid profiles and increase liver glycogen content through glycogen synthase kinase 3. Phytother Res 2010; 24:220-224.

32. Jeong W, Jeong D-H, Do S-H, Kim Y-K, Park H-Y, Kwon O-D, et al. Mild hepatic fibrosis in cholesterol and sodium cholate diet-fed rats. J Vet Med Sci 2005; 67:235-242.

33. Beltroy EP, Liu B, Dietschy JM, Turley SD. Lysosomal unesterified cholesterol content correlates with liver cell death in murine Niemann-Pick type C disease. J Lipid Res 2007; 48:869-881.

34. Wang D, Wei Y, Pagliassotti MJ. Saturated fatty acids promote endoplasmic reticulum stress and liver injury in rats with hepatic steatosis. Endocrinology 2006; 147:943-951.

35. Abo Salem OM, Abd Ellah MF, Ghonaim MM. Hepatoprotective activity of quercetin against acrylonitrile induced hepatotoxicity in rats. J Biochem Mol Toxicol 2011; 25:386-392.

36. Domitrović R, Jakovac H, Vasiljev Marchesi V, Vladimir-Knežević S, Cvijanović O, et al. Differential hepatoprotective mechanisms of rutin and quercetin in CCl4-intoxicated BALB/cN mice. Acta pharmacol Sin 2012; 33:1260-1270.