THE INFLUENCE OF COPTISINE BISULFATE ON THE EVOLUTION OF ACUTE TOXIC HEPATITIS

In the experiments made on white rats was studied the influence of coptisine bisulfate, alkaloid extracted from Chelidonium majus, in the following doses: 10 mg/kg and 20 mg/kg on acute toxic hepatitis cause by carbon tetrachloride. It was established that the researched substance reduced hepatic cytolysis and cholestasis through reestablishment of the transaminases activity and lactate dehydrogenase, while lowering the alkaline phosphatase/alanine aminotransferase ratio and modulated the deflection of the metabolic parameters of acute toxic hepatitis. Coptisine bisulfate corrected the carbon tetrachloride caused hypoproteinemia when administered for 7 days and normalized the albumin level at 14th day of treatment of acute toxic hepatitis. KEYWORDS


Introduction.
Hepatoprotective activity of Chelidonium majus shows benefic influence on the liver functions due to isoquinoline derivatives with antioxidant, choleretic, anti inflammatory and immunomodulatory effects, one of them being coptisine [2,5]. Chelidonium majus ethanolic extract proved the capacity of reducing the hepatotoxicity induced by carbon tetrachloride, through lowering the lipids cumulation, celular necrosis grouth and the lack of [6].
A certain amount of clinical studies that contain extracts from the patients with biliary tract and gallbladder disorders (gallstones, cholecystitis, cholangitis, postcholecystectomy syndrome, alcoholic toxic hepatitis) sugested a significant improvement of the clinical, instrumental and laboratory parameters (bilirubin, transaminases, complete blood count) [6].
Scientific literature analisis allow us to conclude that extract and alkaloids of Chelidonium majus manifests hepatoprotective activity through: decreasing the production of the reactive species of Oxygen; inhibition of the lipids peroxidation; glutathione synthesis rise and antioxidant enzyme activity; reduction of the inflammatory process due to decreasing of proinflammatory cytokines production; correction of lipidic metabolism disorders.
Due to the lack of coptisine bisulfate toxicity [3,4], compared with berberin and other alkaloids of Chelidonium majus and considering the hepatoprotective effect, it was established the aim of experimental research of coptisine bisulfate on acute toxic hepatitis (ATH) shaped in white rats by administering carbon tetrachloride.
Results. Using coptisine bisulfate in the treatment of ATH, there were studied the influence of this alkaloid on the evolution of acute hepatic lesion when using 10 and 20 mg/kg strenghts for establishing a strengh-hepatoprotective effect dependency.
When ATH was shaped at the 7th day was determined a decreasing of the acidic phosphatase from 4,8±0,3 UI/l in the witness lot to 3,7±0,3 UI/l (P<0,05), which was maintained at the same level at 14th day as well. When coptisine bisulfate was administered for 7 days in the doses 10 mg/kg and 20 mg/kg it was established a significant reduction of the acidic phosphatase level in the witness lot, and in the experimental hepatitis one as well. When the alkaloid was used for 2 weeks in both strenghts the acidic phosphatase content increased from 3,7±0,3 UI/l in the lot with CCl 4 to 4,2±0,4 UI/l at the dose of 10 mg/kg and 4,1±0,3 UI/l at the dose of 20 mg/kg (P>0,05).

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Tabel 2. The modification of alkaline phosphatase activity, acidic phosphatase and GGTP in serum in carbon tetrachloride induced HT and the coptisine bisulfate 10 and 20 mg/kg usage In ATH, induced by CCl 4 , activity of GGTP did not change significantly at the 7th day, but increased a lot at the 14th day (tab. 2). When coptisine bisulfate was administered in the dose of 10 mg/kg the level of the enzyme increased from 3,6±0,3 UI/l in the lot with CCl 4 , to 4,3±0,7 UI/l (P> 0,05), while the alkaloid in the dose of 20 mg/kg was decreasing snonsignificantly the content of GGTP (tab. 2). The usage of the alkaloid for 2 weeks contributed to diminishing of the GGTP level from 6,7±0,7 UI/l int he lost with experimental hepatitis, to 4,1±0,4 UI/l and 4,2±0,5 UI/l (P<0,05) in doses of 10 mg/kg and 20 mg/kg accordingly.
In the process of shaping the experimental hepatitis at the 7th day there was noticed a tendency of growing the bilirubin total level in rats from 40,1±2,6 mmol/l to 44,2±3,6 mmol/l (P>0,05), direct bilirubin from 23,0±1,4 mmol/k to 26,3±2,7 mmol/l (P>0,05) and indirect bilirubin from 17,1±1,3 mmol/l to 17,9±1,6 mmol/l (P>0,05). When coptisine bisulfate was administered in the doses of10 mg/kg and 20 mg/kg for 7 days there were no main changes in the total bilirubin content and its fractions (tab. 3). After 14 days there were no significant diversion in total, direct and indirect bilirubin level in comparison with the witness lot, in animals with ATH. Coptisine bisulfate, used for 2 weeks, both strenghts, did not influence the content of bilirubin and its fractions (tab. 3). Tabel 3. The modification of the total, direct and indirect bilirubin level in serum in carbon tetrachloride induced HT and coptisine bisulfate 10 and 20 mg/kg usage

Animal groups
Nr. Of animals When determining the LDH level at the 7th day after shaping ATH it was determined a growth of LDH from 386,2±12,8 UI/l in the witness lot to 1005,7±72,5 UI/l (P 1-2<0,05 ), following to grow until 1355,7±83,7 UI/l (P<0,05) in the 14th day. Whencoptisine bisulfate in strenghts of 10mg/kg and 20mg/kg was administered for 7 and 14 days, the LDH level was at the position of the animals in the witness lot (tab. 4).
When shaping ATH it was noticed a non esential fall of the glucose level in the 7th day and a non significant increase in the 14th day. Coptisine bisulfate, in 10 mg/kg and 20 mg/kg doses, nearly did not influence after 1 week and after 2 weeks as well, the changes of blood glucose induced by the hepatotoxic substance (tab. 4).  Carbon tetrachloride did not influence the cholesterol total level in the 7th day, but determined a increase of it in the 14th day from 5,09±0,04 mmol/l in the witness lot to 5,28±0,05 mmol/l (P<0,05). Coptisine bisulfate (10 mg/kg and 20 mg/kg), administered for 7 and 14 days determined a decrease of the total cholesterol content (tab. 5).
In the experimental hepatitis were not ascertained esential changes of the total protein level, both in 7th and 14th day (tab. 5). In the same time the hepatotoxic determined a decrease of the albumin content from 5,28±0,05 g/l in the witness lot to 30,6±0,8 g/l (P<0,05). Coptisine bisulfate in doses of 10 mg/kg and 20 mg/kg, when administered for 7 days, corrected the CCl 4 induced hypoproteinemia. The albumin level in the 14th day in animals with ATH and the ones treated with coptisine bisulfate was at the level of the witness lot (tab. 5).

№ 8(36), Vol.2, August 2018
Tabel 5. The modification of the cholesterol level, total proteins and albumin in serum in carbon tetrachloride induced HT and coptisine bisulfate 10 and 20 mg/kg usage According to the obtained results, coptisine bisulfate, one of Chelidonium majus alkaloids is a component that lacks in systemic toxicity, owns hepatoprotective properties through re-establishing the transaminases activity, reducing the Ritis coefficient and normalizing the cholestasis in acute toxic hepatitis induced by carbon tetrachloride. This fact indicates its contribution to pharmacological effects of celandine extracts and the possibility of using of this product in the complex treatment of toxic hepatitis.

Conclusions.
1. Coptisine bisulfate, in doses of 10 mg/kg and 20 mg/kg, in a dose dependent way, annihilated the cytolisis syndrome by reestablishing the transaminases activity, esspecially ALAT, and the obvious decrease of the Ritis ratio.
2. It was established the reduction of cholestasis under the influence of coptisine bisulfate by non esential growth of alkaline phosphatase activity and of total, direct and indirect bilirubin level, (decrease of FA/ALAT ratio) and modulated the metabolic parameters deflection of acute toxic hepatitis.
3. When administered for 7 and 14 days, coptisine bisulfate in strenghts of 10 mg/kg and 20 mg/kg, normalises the LDH content at the level or witness lot animals, but the blood glucose level induced by the hepatotoxic substance was not influenced. 4. Coptisine bisulfate corrected the hypoproteinemia caused by CCl 4 , when administered for 7 days and normalised the albumin level in the 14th day of treatment of the ATH animals.