Issues in Biological Sciences and Pharmaceutical Research Vol.4(1),pp.1-5 January 216 Available online at http://www.journalissues.org/ibspr/ http://dx.doi.org/1.15739/ibspr.16.1 Copyright 216 Author(s) retain the copyright of this article ISSN 235-1588 Original Research Article Hepatoprotective effect of a traditional drug, gurgum-7 on carbon- tetrachloride-induced liver damage Received 27 November, 215 Revised 18 December, 215 Accepted 23 December 215 Published 22 January, 216 Amarzaya D 1, Uuganbayar B 1, Dejidmaa B 1, Adilzaya D 2, Chimedragchaa CH 1, Tsend- Ayush D 2, Dagvatseren B 1 and Tsend-auysh G 3 1 Institute of Traditional Medicine and Technology of Mongolia. 2 Health Sciences University of Mongolia. 3 Zadgai Tsagaan Rashaan Subilal *Corresponding Author E-mail: amarzaya_dj@yahoo.com Tel: 976-11-34313 In recent years, one of the leading diseases in the Mongolian population is digestive system diseases among which increased incidence of liver disease in the population is becoming a most challenging situation as it is one of the leading causes of mortality. As of 213, within 5 leading causes of morbidity in a population of 1., digestive system disease stands 2 nd while liver cirrhosis is ranked 3 rd in the leading causes of mortality. Therefore, the Mongolian traditional drug was selected to determine its impact on liver damage caused by carbon tetrachloride compared with the standard group which used. The study thus shows that prescription showed hepatoprotective effect. Key words:, CCI4, hepatoprotective effect. INTRODUCTION In recent years, the use of traditional medicine, especially herbal medicine and its substances is increasing rapidly. A recent survey shows that synthetic drugs/chemical drugs/consumption have increased by 2.1-2.5%, while natural drugs/herbal medicine has increased by 3.5-4.8% worldwide. WHO has published several guidelines (sustainable/rational use for medicinal plants) relating to the above issue. These guidelines recommended an appropriate use of natural SHC substances (raw materials) and herbal medicines to reduce its side effects in order to improve its safety and effectiveness (RSTMWP). In last several years, one of the leading causes of morbidity per 1, populations was digestive system diseases. Among them, liver diseases stand as one of the most common causes of mortality which requires special attention. As of 213, digestive system diseases stand 3 rd of 5 with leading causes of morbidity per 1, populations, while liver cirrhosis stands 3rd for the mortality caused by diseases (Uranbaigali et al., 215). Therefore, based on the above statistic data, we studied the influences of 2, a traditional herbal prescription which has been commonly used in practice to eliminate the heat of the liver. The purpose of the study was to determine the impact of a traditional drug, on carbon tetrachloride-induced liver damage in Wistar rats. The aims of the study are to carry out biochemical and histological analyses and comparative study of impacts on chronic liver disease. MATERIALS AND METHODS The present study was carried out in the Pharmacology Laboratory of the Institute of Traditional Medicine and Technology with the support of the pathological histology departments of the medical school of the University of Medical Science, Ulaanbaatar city, Mongolia. 7 Wistar albino rats (18-25 g) of either sex were used for the experiment. They were kept in standard animal cages and maintained at room temperature (2 ± 2 C) and relative humidity (55 ± 5%) under 12 h of light and dark cycle. The animals were allowed free access to commercial animal feeds (pelleted feed) and water. Experiments were carried out according to the rules of procedure with the use of experimental animals. The study protocol was approved by
u/l Issues Biol. Sci. Pharma. Res. 2 Table1.The model of hepatotoxicity for experimental animals Groups n Treatment group 1 2.5 ml distilled water was given orally for each day. Control group 2 5% CCl4 (.4ml/1gm) subcutaneously injection once per day, for 4 day 2.5 ml distilled water was given orally for each day. Live-52 2 5% CCl4 (.4ml/1gm) subcutaneous injection once per day, for 4 days. 1 mg/kg Liv 52 4.5 was given orally each day for 28 days Gurgum 7 2 5% CCl4 (.4ml/1gm) subcutaneous injection once per day, for 4 days. 275 mg/kg Gurgum 7 was given orally each day for 28 days 2 15 1 ALT Control 5 Figure 1: s impact on experimental animal liver damage by serum level of ALT the Ethics Committee of the Health Sciences University of Mongolia. The acute toxicity study The animals were divided into 3 groups (n=2). 5% carbon tetrachloride (CCl 4) was subcutaneously injected at.4 ml/1 gm dose for 4 days (Table 1) and the evaluation taken on days 3, 7, 14 and 28 (Figure 1) (Khishigjargal et al., 26); Ambaga (1992). The study results were obtained by evaluating aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) in serum level and histomorphological examination. Results are presented as mean, standard deviation (SD) and standard error (m) at 95% confidence interval. Statistical analysis was performed using one-way analysis of variance (ANOVA). P value <.5 was considered significant. RESULTS In the group treated with (study group), serum ALT level decreased by 22.3% in 3 days, 37% in 7 days, 61.2% in 14 days and 49.2% in 28 days (Figure 2) compared to the control group (p>.1). There were statistically significant differences in serum AST level between the healthy and control groups. Serum AST level was eliminated by 39.2% in 28 days compared to the control group (p>.1). According to Figure 3, serum ALP level in the study group was lowered by 44.5% in 7 days, 52.1% in 14 days and 39.3% in 28 days respectively, compared to the control group. (p>.1) Comparative histological analysis was carried out on the liver tissues of experimental animals. According to Figure 4A, in the control group CCI 4 induced liver tissue damage showed inflammatory cells around the hepatic cords and triads, large and small fatty changes around the central vein, hepatocytes necrosis and decomposition, dismissed pillar structure of hepatocytes and blood stasis in the sinusoids in 3 days. Meanwhile, in Figure 4B, liver tissue showed small fatty changes, mild protein changes in hepatocytes and micro vascular blood stasis but the liver pillar structure remained. In Figure 5A, CCI 4 induced control group liver tissue showed diffused fatty droplets, blood stasis, liver pillar and lobular structure was lost in lobular center, HCC formation of protein fatty metamorphosis in 7 days.
u/l u/l Amarzaya et al. 3 4 3 2 AST Control 1 Figure 2: Gurgum 7 s impact on experimental animal liver damage by serum level of AST 5 4 3 2 ALP control 1 Figure 3: Gurgum 7 s impact on experimental animal liver damage by serum level of ALP Figure 4: CCI4 induced liver damage histopathology in 3rd day. (A: control group H&Ex1; B: H&Ex1 ) a. artery b. vein c. Liver Trinity d. inflammatory cell infiltration e. fatty metamorphosis j. HCC regeneration, F. blood stasis i. HCC decomposition and necrosis
Issues Biol. Sci. Pharma. Res. 4 Figure 5: CCI4 induced liver damage histopathology in 7 days. (A-Control-H&Ex1; B-, H&Ex2) b. vein, c. liver triads, d. inflammatory cell infiltration. e. fatty changes; j. HCC compensation calving: Figure 6: CCI4- induced liver damage histopathology in 14 days. (A-Control-H&E1x1, 2x2; B- H&Ex2) b. vein s. liver triads d. inflammatory cell infiltration. e. fatty changes J. HCC compensation calving Meanwhile Figure 5B showed that in the -treated group, pillar structure was stored, few lymphocytes and mono-nucleus surrounded the triads, mild fatty changes in the lobular edges and multiplied HCC compensation calving (nucleus fragmentation) in the examining field within 7 days. Figure 6A showed histology in the control group in 14 days. It showed hepatocellular necrosis and degeneration, blister with decomposition and dismissed pillar structures. Blood stasis in the vein and micro-vascular, arterial plasma sedimentation occurred in the valley. An Inflammatory infiltration found in portal triads. In Figure 6B, treated group showed that pillar structure was stored within 14 days and HCC regenerative changes was observed/increased amount of dual core cells in 28 days. Figure 7A showed HCC epicenter necrosis, granulamatous protein changes. Veins and micro blood vessels were dilated with not much blood flow in it; valley was empty. Figure 7B showed that in treated group, liver pillars structure was maintained and sinusoids and capillary structures were normal in 28 days. DISCUSSION CCI 4 can damage HCC through oxidative mechanisms, changes in connective tissue structures which can further cause degradation of liver function. Besides hepatocellular regeneration and inflammatory infiltration, hepatocellular regeneration proliferation of hepatic stellate cells and deposition of connective tissue are major features of liver histopathology (Qi-ppeum et al., 25). According to this study, can decrease serum ALT level (which refers to cell decomposition) by 22.3-61.2%, AST by 8.6-39.6%, ALP by 9.1-52.1%, respectively, due to the pathohistology analysis liver pillar structure was maintained, sinusoids and capillary structure was normal and HCC regeneration and inflammatory cell infiltration degenerated. One of the main ingredients in s is Carthamus tinctorius L, which has hepatoprotective effect on CCl4 induced liver damage and at the same time can reduce cirrhosis damage level. Zhang et al. (211) proved that this
Amarzaya et al. 5 Figure 7: CCI4-induced liver damage histology in 28 days (A. Monitoring H & E kh2; B., C-28 days, H & E kh2) effect can be explained by substances called Kartamin (Wu et al., 213; Shijun et al., 213). Carthamus tinctorius L is also known to reduce HCC compensation, decrease cholestasis, connect biliation and cholecyst during hepatitis (Bayarmaa et al., 213). Conclusion The study proves that a traditional recipe () can decrease HCC decomposition and necrosis through lowering serum ALT, AST and ALP levels in experimental animals while simultaneously, its hepatoprotective effect was confirmed by pathohistology analysis. Conflict of Interests The authors declare that there is no conflict of interests regarding the publication of the paper REFERENCES Amarzaya D, Altanchimeg A, Chimedragchaa Ch, Tsend- Ayush, Dagvatseren B (215). Influence of traditional ingredients of Nephric on the renal failure induced with kanamycin on the rat. Mongolian J. Health Sci. 12(1):29-33. Amarzaya D, Dagvatseren B, Chimedragchaa Ch, Tsend- Ayush D (215). Systematic traditional medicine and special characteristic to compound module-hierarchical drugs. J. Mongolian Traditional Med., 4(1):43-48. Ambaga M, Chültemsüren M, Sarantsetseeg B. (1992) To explore and customize hepatoprotective new recipes based on the Oriental medicine theory. J. Med. Mongolia 2(8):39-46. Bayarmaa M, Badamsuren D, Tserendagva D (213). Effect of Dracocephalum foetidum L. on isoniazid induced liver injury. Mongolian J. Health Sci., 23 (9):32-34. Khishigjargal S, Sarantsetseg B (26). Hepatoprotective effect of Saxifraga hirculus L. extract. The 7th Mongolian Russian International Medical Symposium. pp. 38-4. Qi-ppeum l, Won-il J, Da-hee J, Sun-hee D, Tae-hwan K, Kyushik J (25). Diagnostic Evaluation of Carbon Tetrachloride-induced Rat Hepatic Cirrhosis Model., Anticancer Res. 25:129-138. Shijun Y, Yuping T (213) Chemical and Biological Properties of Quinochalcone C-Glycosides from the Florets of Carthamus tinctorius. Molecules., 18:1522-15254. Siddhartha S, Archana M, Pradeep M (211). Hepatoprotective activity of Cajanus cajan against carbon tetrachloride induced liver damage. Int. J. Pharma. Pharma. Sci. 3(2):146-147. Syed AB, Iqbal MM, Kiranmai M, Ibrahim M (212). Hepatoprotective activity of phyllanthus amarus seeds extracts in CCl4 treated rats: In vitro & In vivo, Global J. Med. Res., 12(6):41-42. The Regional Strategy for Traditional Medicine in the Western Pacific. (211-22).pp.11-14. Uranbaigali N, Ariunaa Z, Chimedragchaa C (215) Hepatoprotective effect of Antomen preparation on CCI 4 induced hepatic damage in rats. J. Oriental Med., 1(9):71-74. Wu SC, Yue, Ding H (213) Carthamus red from Carthamus tinctorius L. exerts antioxidant and hepatoprotective effect against CCl4-induced liver damage in rats via the Nrf2 pathway. J. Ethnopharmacol. pp.148-158. Zhang YB, Guo J, Dong HY, Zhao (211) Hydroxysafflor yellow A protects against chronic carbon tetrachlorideinduced liver fibrosis. Eur. J. Pharm., 66:438-444.