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6TMR Modern Herbal Medicine  2019, Vol. 2 Issue (3): 121-130    DOI: 10.12032/TMRmhm2017A49
Orginal Article     
The Protective Effect of Jiujiuguiyi, a Medicine and Food Homologous Formula, on Acute Alcohol Poisoning Mice
Yang-Fan Zhou1,2,3, Tian-Ze Zhang1,2,3, Si-Qi Deng1,2,3, Meng Zhang1,2,3, Yu-Jiao Zhan1,2,3, Yi-Fang Li1,2,3,*(), Rong-Rong He1,2,3,*()
1Guangdong Province Research and Development Center for Chinese Medicine in Disease Susceptibility, College of Pharmacy, Jinan University, Guangzhou, China.
2International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry of Education (MOE), College of Pharmacy, Jinan University, Guangzhou, China.
3Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research,College of Pharmacy, Jinan University, Guangzhou, China.
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Abstract

Background: Nowadays, acute alcoholic intoxication has become the third public problem in China, and the anti-inebriation products mainly aimed at increasing the activity of enzyme involved in the alcohol metabolism, which is a single mechanism that can accelerate alcohol metabolism. Thus, a new formula, Jiujiuguiyi (JJGY) which could protect liver, relieve the abnormal excitability of the center and improve muscle retardation at the same time is designed by us. Methods: The model of acute alcoholic intoxication was established by intragastric administration with 0.12 ml/10g 50% alcohol in mice. JJGY was orally administrated (gavage) once a day for 20 consecutive days before the establishment of acute alcoholic model. Mice were randomly divided into 8 groups with 8 each: blank control group (CON), model group (M), Haiwangjinzun positive control group (HWJZ), experimental groups (AL, AH, BL, BH, AB). Giant, crawling time on the rota-rod, the activities of aspartate amino trans- ferase (AST), alanine amino transferase (ALT) and superoxide dismutase (SOD) in both liver and serum, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), glutathione peroxidase (GSH-PX) in liver as well as the HE staining of liver slices, the formation of malondialdehyde (MDA) in serum were determined after acute alcoholic intoxication. Results: Compared with model group, JJGY significantly decreased the AST and ALT activity in liver and serum and MDA activity in serum. Meanwhile, it enhanced the ADH and ALDH level in liver as well as the hepatic and serous SOD activity, indicating more efficient metabolism of alcohol and less hepatic injury. HE staining results also proved that JJGY could reduce alcoholic liver cell injury, and the effect was more obvious in the group medicated before alcohol administration. Moreover, JJGY significantly prolonged the crawling time on the rota-rod and improved the gait of mice and the effect was proved to be better than the widely used health product Haiwangjinzun. Conclusions: This study suggests that JJGY is able to protect liver, relieve the abnormal excitability of the center and improve muscle retardation after acute alcoholic intoxication. Its liver protection effect is likely related to its modulation on the alcohol metabolizing and antioxidant enzymes.

Highlights

In the present study, a new compound Chinese herbal medicine formula, Jiujiuguiyi, was designed by using the medicine and food homology theory. The formula aims at protecting liver, relieving the abnormal excitability of the center and improving muscle retardation at the same time. Acute alcoholic intoxication model in mice was built, then the ethology test and biochemical test were conducted to exam the efficacy of the formula in different preparations. The results suggest that JJGY can protect the acute alcoholic intoxication mice through multiple mechanisms, providing a new way to develop antialcoholismic drug homologous food.



Key wordsJiujiuguiyi (JJGY)      acute alcoholic intoxication of mice      medicine and food homology      ethology test     
Published: 25 July 2019
Fund:  This work was Financial supported by China College Students' Innovation and Entrepreneurship Project (Grant NO. CX18012 and 201910559129S)
Corresponding Authors: Li Yi-Fang,He Rong-Rong     E-mail: rongronghe@jnu.edu.cn;liyifang706@jnu.edu.cn
About author: # Both authors contributed equally to this work.
Cite this article:

Yang-Fan Zhou, Tian-Ze Zhang, Si-Qi Deng, Meng Zhang, Yu-Jiao Zhan, Yi-Fang Li, Rong-Rong He. The Protective Effect of Jiujiuguiyi, a Medicine and Food Homologous Formula, on Acute Alcohol Poisoning Mice. 6TMR Modern Herbal Medicine, 2019, 2(3): 121-130. doi: 10.12032/TMRmhm2017A49

URL:

https://www.tmrjournals.com/mhm/EN/10.12032/TMRmhm2017A49     OR     https://www.tmrjournals.com/mhm/EN/Y2019/V2/I3/121

Figure 1. Effect of JJGY on hepatic AST, ALT, ADH, ALDH, GSH-PX and SOD
The livers were collected from acute alcohol poisoning mice after ethology test, then they were homogenized at 1:9 in cold normal saline (NS), and centrifuged at 3000 rpm for 15 min, and the supernatant was collected for the analysis of transaminase (AST, 1a), alanine aminotransferase (ALT, 1b), glutathione peroxidase (GSH-PX, 1e), superoxide dismutase (SOD, 1f), alcohol dehydrogenase (ADH, 1c) and aldehyde dehydrogenase (ALDH, 1d) with Mk3 fluorescence microplate System. Compared with control group, ** P < 0.01, *** P<0.001, compared with model group, # P < 0.05, ## P < 0.01, ### P < 0.001.
Figure 2. Effect of JJGY on hepatic tissue
Mouse livers from different groups were fixed in 10% formalin solution and processed by hematoxylin-eosin (HE) staining. Conventional HE staining sections were prepared and histopathological changes were observed under optical microscope.
Figure 3. Effect of JJGY on serum AST, ALT, MDA and SOD
Whole blood was collected from the orbit of acute alcohol poisoning mice after ethology test. The blood was centrifuged at 2500 rpm for 15 min and the serum was collected. The activities of serum aspartate transaminase (AST, 3a), alanine aminotransferase (ALT, 3b), malondialdehyde (MDA, 3c) and superoxide dismutase (SOD, 3d) were analyzed with Mk3 fluorescence microplate System.
Figure 4. Effect of JJGY on motor coordination.
After setting up the acute alcoholic intoxication model, the mice were placed on a rota-rod analyzer to record the Falling Time (4a) after adapting the environment. After plenty of rest, the mice were placed in the catwalk analysis system for recording Candence (4b) and Swing Speed (4c) after adapting the environment. Compared with control group, ** P < 0.01, *** P < 0.001, compared with model group, # P < 0.05, ## P < 0.01, ### P < 0.001.
Figure 5. Effect of JJGY on gaits of mice.
After motor coordination test, mice got plenty of rest. Then, the mice were placed in the catwalk analysis system for recording Run Duration (5a), Walking Speed (5b), Step Cycle (5c), Timing View (5d, RF, right front; RH, right hind; LF, left front; LH, left hind) and Footfall Patterns (5e, Aa, RF-RH-LF-LH; Ab, LF-RH-RF-LH; Ca, RF-LF-RH-LH; Cb, LF-RF-LH-RH; Ra, RF-LF-LH-RH; Rb, LF-RF-RH-LH). The giant of mice should have a certain regulation.
Compared with control group, ** P < 0.01, *** P<0.001, compared with model group, # P < 0.05, ## P < 0.01, ### P<0.001.
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