Protective Effect of Diterpene Phenol Extract of Rosemary on Several Liver Injuries

 Abstract : To observe the effects of rosemary diterpene phenol extract on liver damage induced by carbon tetrachloride (ccl4), thioacetamide (TAA), acetaminophen (AAP), ethanol (alcohol) and DL-ethionine (DL-E) hepatotoxicity in mice. METHODS The liver toxicants were administered to mice after ig administration of rosemary diterpene phenol extract, and the serum alanine aminotransferase (ALT) activity and hepatic triglyceride (TG) content were measured. Results Rosemary diterpene phenolic extracts significantly inhibited the elevation of ALT and TG induced by ccl4, TAA, AAP, alcohol, and DL-E, and had no significant effect on ALT and TG in normal mice. The LD5O of ig rosemary diterpene phenol extract in mice was more than 15g/kg. Conclusion Rosemary diterpene phenol extract has hepatoprotective and enzyme-lowering effects, and has the prospect of development and application.

 

Diego Rosmarinus officinalis L. is a genus of rosemary in the Labiatae family. It is native to Mediterranean countries, and has been successfully introduced into cultivation on a large scale in Dushan County, China under the guidance of experts from the Institute of Botany of the Chinese Academy of Sciences in recent years. Rosemary diterpene phenolic components have obvious antioxidant, anti-tumor, anti-HIV and microbial activities[1] , but studies on the hepatoprotective and hypomelanotic effects of rosemary diterpene phenols have not been reported. We observed that rosemary diterpene phenolic extracts had significant protective effects against liver injury caused by several commonly used chemical toxicants.

 

1 Material

The powder of rosemary diterpene phenol extract (2Og per gram of raw herb) was supplied by Associate Researcher Wu Huamei and Visiting Scholar Zhang Gao from Guizhou Institute of Biology. In the experiment, the powder was solubilized with O.2 mL of Tween-8O, and then made up to the required dosage with distilled water. The ccl4 was dissolved in vegetable oil. Acetaminophen (AAP), the product of Beijing No.2 Pharmaceutical Factory, prepared with distilled water, and dissolved by heating. Biphenyl dibenzoate (BDD), produced by Zhejiang Xinchang Pharmaceutical Factory, supervised by the Institute of Pharmaceuticals, Chinese Academy of Medical Sciences, batch number 9O1222, was supplied by the Animal Section of Guizhou Epidemic Prevention Station.

 

2 Methodology and results

2. 1 Effect of rosemary diterpene phenol extract on ccl4-induced liver injury in mice.

The mice were randomly grouped into 1O mice per group, normal control group, and 3 administration groups ig rosemary diterpene phenol extract 5O, 1OO, 2OO mg/(kg . d × 4d, positive control group ig biphenyl dibenzoate 2OO mg/(kg . d × 4d), respectively. d) × 4d, positive control group ig biphenyl dibenzoate 2OO mg/(kg . d) × 4d, the last dose of biphenyl dibenzoate 2OO mg/(kg . d) × 4d. d) × 4d, 8h after the last dose, ip O.1%ccl4 vegetable oil solution 1OmL/kg, followed by fasting, 16h later, blood was taken from severed head, and the serum was separated for the determination of ALT according to the law[2] ; and a small piece of liver tissue was made into 5% liver homogenate with ice-cold PBS (O.1mol/L, pH7.4), and TG was measured according to the law[3] , and the results are shown in Table 1. The results showed that the extract of rosemary diterpene phenol had a significant and dose-dependent lowering effect on the elevation of ALT induced by ccl4, and also had a significant lowering effect on the elevation of TG.

 

2.2 Effects of rosemary diterpene phenolic extract on thioacetamide (TAA), acetaminophen (AAP) and alcohol-induced liver injury in mice.

The experimental animals were treated in the same way as in item 2.1. ig Rosemary diterpene phenol extract 1OOmg/(kg . d) × 5d, 8h after the last dose, ip TAA 5Omg/kg, ip AAP175mg/kg, ig 45% alcohol 1OmL/kg, the results are shown in Table 2. It can be seen that rosemary diterpene phenol extracts have a significant effect on the elevation of ALT and hepatic TG induced by TAA and AAP, and have a significant effect on the accumulation of hepatic fat induced by alcohol in mice. The effect of rosemary diterpene phenol extracts on the elevation of ALT and hepatic TG induced by TAA and AAP was shown in Table 2.

 

2. 3 Effect of rosemary diterpene phenol extract on DL-ethionine (DL-E)-induced fatty liver in mice.

After grouping the mice, the drug group was given ig rosemary diterpene phenol extract 1OOmg/(kg . d) × 5d, 1h after the last dose, and the experimental group was given ig DL-E25Omg/kg once. d) × 5d, 1h after the last dose, animals in the experimental group were given ig DL-E25Omg/kg once, and the hepatic TG content was measured after 24 h. The results are shown in Table 2.

Phenol extracts significantly inhibited DL-E-induced TG in animal liver.

 

2.4 Effect of rosemary diterpene phenol extract on ALT and hepatic TG in normal mice :.

The animals were randomly divided into two groups of 10 animals each, the normal control group and the drug group ig 1OOmg/(kg.d.) of diterpene phenol extract, and were fasted for 24h after the last dose of the drug. In the normal control group, 1O animals were randomly divided into two groups, each with 1O animals, the normal control group was given 1OOmg/(kg.d) × 4d, and the animals were fasted for 24h after the last dose of the drug, and the animals were put to death to take the serum test of ALT, and the liver test of TG, the results were: ALT in the normal control group was (155. 8±1OO. 8)U/1OOmL, and that of the rosemary group was (131. 6±1OO. 9)U/1OOmL (P>O. O5), and that of the normal control group was (O. 26± O. 18)mmol/L. (O. 26± O. 18) mmol/L in the normal control group and (O. 19± O. O5) mmol/L in the rosemary group (P>O. O5). The diterpene phenolic extract of rosemary had no significant effect on ALT and TG in normal animals.

 

2. 5 Rosemary diterpene phenol extract Acute toxicity test.

2O mice were taken, once ig rosemary diterpene phenol extract 15g/kg, observed for 7 d, no abnormal performance and death were observed, suggesting that the LD5O of rosemary diterpene phenol extract is greater than 15g/kg, and the toxicity is very small.

 

3 Discussion

In this study, it was observed that the hepatoprotective effect of the extract of rosemary diterpene phenol on liver injury caused by ccl4, TAA, AAP, alcohol, and DL-E in mice was favorable. The mechanisms of liver damage are different for all hepatotoxicants, and it can be concluded that the protective effect of rosemary diterpene phenol extracts on the liver is not realized by a single protective mechanism. It was also observed that the extract of rosemary diterpene phenol had no significant effect on ALT activity and liver TG content in normal animals, which indicates that rosemary diterpene phenol does not have a direct inhibitory effect on ALT, but does have a protective effect on the liver, and its toxicity is very small, which suggests that rosemary diterpene phenol is safe for consumption, and it is very promising to be developed and utilized as a new hepatoprotective and enzyme-lowering drug.

 

References.

[1] TU Pengfei , XU Zhanhui , ZHENG Jiatong , et al.  Chemical composition and application of rosemary [J].  Natural Products Research and Development , 1998, 1O(3): 62-68.

[2] Shanghai Medical Laboratory .  Experimental Clinical Testing [M].  Shanghai : Shanghai Science and Technology Publishing House , 1965.

[3] Shanghai Sixth People's Hospital Laboratory Group.  Simple and rapid method for the determination of serum triglycerides [J] , Medical Information Exchange , 1975, (5) :41.