Nghiên cứu độc tính và tác dụng bảo vệ gan của sản phẩm vismisco trên thực nghiệm ttta

MINISTRY OF EDUCATION & MINISTRY OF TRAINING HEALTH HANOI MEDICAL UNIVERSITY BUI THI QUYNH NHUNG STUDY TOXICITY AND THE HEPATOPROTECTIVE EFFECTS OF VISMISCO PRODUCTS ON EXPERIMENT Major: Pharmacology and toxicity Code: 9720118 SUMMARY OF MEDICAL DOCTORAL THESIS HANOI - 2023 THIS STUDY WAS COMPLETED AT HANOI MEDICAL UNIVERSITY Supervisors: Assoc.Prof Dr Pham Thi Van Anh Assoc.Prof.Dr Nguyen Trong Thong Reviewer 1: Reviewer2: Reviewer3: The thesis is defended in front of the University Thesis AssessmentCouncil More information of the thesis could be found at: THE AUTHOR’S SCIENTIFIC RESEARCHES Bui Thi Quynh Nhung, Nguyen Van Son, Pham Thi Van Anh (2016) Anti-inflammatory effect of Vismisco on experimental research Journal of Vietnamese Medicine, 449, 16-24 Bui Thi Quynh Nhung, Nguyen Van Son, Pham Thi Van Anh (2018) Effect of Vismisco in mouse liver injury induced by paracetamol Journal of Vietnamese Medicine 472, 569 - 574 Bui Thi Quynh Nhung, Nguyen Van Son, Pham Thi Van Anh, Nguyen Thanh Ha (2019) Evaluation of subchronic toxicity of Vismisco in experimental animals Journal of military pharmaco-medicine, 4, 90 - 97 Bui Thi Quynh Nhung, Pham Thi Van Anh, Nguyen Trong Thong, Luong Ngoc Cuong, Mai Phuong Thanh (2023) Antifibrotic effects of vismisco on carbon tetrachloride-induced liver cirrhosis in mice Journal of Practical Medicine, (162), 272-279 A: INTRODUCTION Rationale A liver is a large organ of the body, responsible for many important and complex functions, and plays an important role in the detoxification and metabolism of substances Causes of liver diseases include bacteria, viruses, parasites, alcohol, drugs or toxic chemicals, which can result in acute hepatitis, chronic hepatitis, possibly progressing to cirrhosis or cancer, when entering the liver In the treatment process of acute and chronic hepatitis, in addition to specific treatments, medicine that acts as a therapeutic agent has been shown to play an important role Liver diseases account for large morbidity and mortality rates in the world According to the 2019 World Burden of Disease report, liver disease accounts for about millions deaths each year In the treatment of acute and chronic hepatitis, in addition to specific treatment measures, drugs with therapeutic effects have been proven to play an important role Vismisco extracted from Vigna radiata (L.) Wilczek, Smilax glabra roxb and Scoparia dulcis L which are three herbs that have been studied and widely used in folk medicine to treat many different diseases We carry out this thesis to demonstrate the scientific basis of the use of Vismisco of liver diseases according, towards the possibility of widely using Vismisco for the clinical treatment of hepatitis and cirrhosis Objectives and contents of the thesis 2.1 Objectives: Studying the acute and semi-chronic toxicity of Vismisco in experimental animals Studying the effects on anti-acute liver damage, anti-chronic hepatitis and antioxidant effects, evaluateacute and chronic anti-inflammatory effects of Vismisco 2.2 Contents: 2.2.1 Studying toxicity - Determining the acute toxicity of Vismisco extracted from Vigna radiata (L.) Wilczek, Smilax glabra roxb and Scoparia dulcis L - Determining the semi-chronic toxicity of Vismisco 2.2.2 Studying the anti-hepatitis, liver injury recover, anti-chronic hepatitis and antioxidant effects of Vismisco - Studying the effects on anti-liver damage (protecting liver and repairing liver damage) of Vismisco - Studying the effects on anti-chronic hepatitis and antioxidant of Vismisco - Studying to the evaluateacute and chronic anti-inflammatory effects of Vismisco New contributions of the thesis - The dissertation is the first study on the toxicity, liver injury recover, antichronic hepatitis and anti-oxidant effects of Vismisco extracted from Vigna radiata (L.) Wilczek, Smilax glabra roxb and Scoparia dulcis L - The results of the study showed that Vismisco extracted from Vigna radiata (L.) Wilczek, Smilax glabra roxb and Scoparia dulcis L has not caused acute toxicity and Vismisco at doses 0.4g/kg/day and 1.2g/kg/day did not produce any toxic signs or evident symptoms of semi-chronic toxicity in experimental animals Vismisco with a dose of 0.6 g/kg/day and 1.8 g/kg/day had hepatoprotective effects and increasing the recovery of liver damage in white mice when toxic by paracetamol 400 mg/kg; Vismisco with a dose of 0.6 g/kg/day and 1.8 g/kg/day had to limit cirrhosis damage and antioxidant in white mice when toxic by long-term CCl4.Vismisco with a dose of 0.4 g/kg/day and 1.2 g/kg/day had an acute antiinflammatory effecton the acute inflammatory model in white sewer rats Vismisco with a dose of 0.6 g/kg/day and 1.8 g/kg/day had a chronic anti-inflammatory effecton the chronic inflammatory model in white house-mice - The results of this study are new contributions of the thesis, which was published for the first time on the toxicity and effects of anti-hepatitis, liver injury recover, anti-chronic hepatitis and antioxidant s in experimentation of Vismisco in Vietnam The scientific and practical significance of the thesis 4.1 The scientific significance of the thesis: The findings of the thesis have contributed to confirming the safety of Vismisco extracted from Vigna radiata (L.) Wilczek, Smilax glabra roxb and Scoparia dulcis L and initially elucidated the effects of acute anti-hepatitis, liver injury recover, anti-chronic hepatitis and antioxidant, creating a premise for further research 4.2 The practical significance of the thesis: It is a scientific basis for the development of effective medicine for the treatment of hepatobiliary diseases from available natural sources The organization of the thesis: The thesis consists of 150 pages, including the following parts: Introduction (2 pages); Overview 41 pages; Subject and research method 15 pages; Research results 41 pages with 29 tables, 29 pictures, charts; Discussion of 48 pages; Conclusions and recommendations (3 pages) The thesis has 174 references in Vietnamese and English B: CONTENT OF THE THESIS Chapter 1: OVERVIEW 1.1 Hepatitis and cirrhosis pathology 1.1.1 Concepts: The word "hepatitis" refers to all diseases that cause degenerative damage, hepatocellular necrosis and damage to the liver's stroma due to an inflammatory response Chronic hepatitis is a liver disease with necrotic and inflammatory lesions, with or without fibrosis, occurring for more than months 1.1.2 Causes: 1.1.2.1 Causes of hepatitis: There are many causes of hepatitis such as viruses, alcohol, poisoning chemicals, etc 1.1.2.2 Causes of cirrhosis: Hepatitis is caused by hepatitis B, C, and D viruses, alcohol, and a number of other causes Among them, viral and alcoholic hepatitis are the main causes of cirrhosis, which account for over 90% of cirrhosis cases 1.1.3 Mechanism of pathogenesis 1.1.3.1 Pathogenesis mechanism of hepatitis - Pathogenesis mechanism of alcoholic hepatitis: In the body, the liver is the most important alcohol metabolite Most alcohol is metabolized in the liver in two stages: Stage 1: metabolizing alcohol to acetaldehyde, Stage 2: Acetaldehyde is formed as a poison, which is quickly converted into acetate by the enzyme acetaldehyde dehydrogenase The metabolic capacity of this stage is limited If acetaldehyde is produced in a large amount, it will not be completely metabolized, causing vasodilation and binding to the cell membrane, which leads to cell damage through toxic, inflammatory and immune - Pathogenesis mechanism of the drug and chemical hepatitis: There are main mechanisms: liver damage due to idiosyncratic reactions and liver damage from the overdose - The role of free radicals in the pathogenesis mechanism of liver damage 1.1.3.2 Pathogenesis mechanism of cirrhosis: The major changes in the mechanism of cirrhosis are progressive diffuse fibrosis of the liver and reorganization of the microcirculation system in the liver The activation of astrocytes (HSC: hepatic stellate cell) increases the proliferation of collagen fibers and the formation of fibrous walls In addition to fibrinolytic activity, the liver cells that survive will increase their proliferative activity to produce regenerating nodules surrounded by fibrous tissue In the end, the liver becomes a fibrous tissue containing liver cell nodules, the microcirculation system that supplies blood to the liver cells and the liver's ability to produce proteins is severely damaged 1.1.4 Liver and oxidative stress * Free radicals * Antioxidants * Antioxidant system in the liver 1.1.5 Some medicinal plants have been studied to treat hepatitis 1.2 A research model of with anti-hepatitis 1.2.1 Research model of using CCl4 causing acute hepatotoxicity 1.2.2 Research model of using paracetamol causing acute hepatotoxicity 1.2.2 Research model of medicine with anti-cirrhosis: long-term use of CCl4 causes cirrhosis of the liver in experimental white mice 1.3 Evaluation methods 1.4 Overview of the Vismisco 1.4.1 Composition of Vismisco: Vismisco is extracted at Department of Pharmacy - Thai Nguyen University of Medicine and Pharmacy meeting basic standards including: Vigna radiata (L.) Wilczek 58.8% Smilax glabra roxb 11.8% Scoparia dulcis L 29.4% Intended dose in human 2.5 g Vismisco/day 1.4.2 Introduction of herbal ingredients of the Vismisco 1.4.3 Studies on biological effects: The domestic and international have researched and demonstrated that Vigna radiata (L.) Wilczek, Smilax glabra roxb and Scoparia dulcis L the has anti-inflammatory, antioxidant, protective and restorative effects of liver damage, anti-hyperlipidemia, Hypoglycemia Chapter 2: RESEARCH METHODOLOGY 2.1 Studied ingredients Vismisco is extracted at Department of Pharmacy - Thai Nguyen University of Medicine and Pharmacy meeting basic standards including: Vigna radiata (L.) Wilczek 58.8% Smilax glabra roxb 11.8% Scoparia dulcis L 29.4% Intended dose in human 2.5 g Vismisco/day 2.2 Medicine, chemicals, machines and equipment for studying: Meets testing standards 2.3 Experimental animals - Healthy white Swiss mice, both breeds, weighing 25.0 ± 2.0 grams (to study the protective and liver damage and anti-cirrhosis recovering effects) and 20.0 ± 2.0 gram (to study acute toxicity, hepatoprotective effect, liver injury recover, antichronic hepatitis and anti-oxidant, anti-chronic inflammatory effects) provided by the National Institute of Hygiene and Epidemiology - Healthy white mice, both breeds, weighing 150 ± 20 grams (to study antiacute inflammatory and semi-chronic toxicity effects) 2.4 Research Methods 2.4.1 Studying toxicity 2.4.1.1 Studying acute toxicity: was conducted under the guidance of the World Health Organization and the Ministry of Health of Vietnam on testing medicine derived from medicinal herbs It has determined LD 50 of test samples on white mice orally according to Litchfield - Wilcoxon method - Before conducting experiments, rats fasted overnight Each batch of 10 white mice is given a study drug sample in increasing doses - Finding the highest non-lethal dose (0%), the lowest non-lethal dose (100%), and intermediate doses and monitoring general condition of mice and the number of deaths in each batch for 72 hours to build a linear graph to determine LD 50 of the sample; then continuing to monitor the general condition of mice until the end of days after taking the study sample 2.4.1.2 Subchronic toxicity of Vismisco study: Subchronic toxicity was studied by using Vismisco samples on white rats orally under the guidance of the World Health Organization Rats received doses of Vismisco: equivalent to 0.4 g / kg / day (equivalent to the expected dose in human) and equivalent to 1.2 g / kg / day (3 times the expected dose per person) for consecutive weeks Rats were evaluated the general condition and change in weight, hematopoietic function, liver and kidney functions before taking the test sample, after weeks, after weeks stopping the sample Liver and kidney histopathology were evaluated after weeks of stoping the sample 2.4.2 Studying the anti-hepatitis, liver injury recover, anti-chronic hepatitis and antioxidant and anti-inflammatory effects of Vismisco 2.4.2.1 Studying anti-hepatitis effects * Studying liver-protection effects: The mice were separated into groups of 15 animals each Group (control): Taking distilled water of 0.2 ml / 10g Group (hepatitis model): Taking distilled water + PAR Group (positive control): Taking silymarin (140 mg / kg / day) + PAR Group (Vismisco dose 1): Taking Vismisco dose equivalent to 0.6 g/kg/day + PAR Group (Vismisco dose 2): Taking Vismisco dose equivalent to 1.8 g/kg/day + PAR The mice continuously took distilled water, silymarin or samples during days On the 8th day, hour after taking the drug (fasted mice 16-18 hours earlier), their livers were damaged in groups to by taking paracetamol orally (PAR) dose of 400 mg / kg, with volume 0.2 ml/10 g 48 hours after being poisoned by PAR, the mice was taken blood of the carotid artery to determine the activity of enzymes AST, ALT and at the same time taken livers to observe histopathology (macroscopic, microscopic) and MDA * Studying the effect of increasing the liver damage recovery of Vismisco: Studying the effect of increasing the liver damage recovery of Vismisco for days: The mice were separated into groups of 10 animals each They had their liver damaged by taking oral dose of PAR 400mg/kg, with a volume of 0.2ml/10g hour after taking PAR, they were given distilled water or reagents corresponding to each batch: Group (control): Taking distilled water of 0.2 ml/10g Group (hepatitis model): Taking PAR + distilled water Group (positive control): Taking PAR + silymarin 140 mg/kg/day Group (Vismisco dose 1): Taking PAR + Vismisco dose equivalent to 0.6 g/kg/day Group (Vismisco dose 2): Taking PAR + Vismisco dose equivalent to 1.8 g/kg/day Mice in groups were given distilled water or reagent for days After days of taking the reagent, they were killed to take carotid artery blood to determine the activity of enzymes AST, ALT; and to take out the liver to MDA Studying the effect of increasing the liver damage recovery of Vismisco for days: Mice were randomly divided into groups, 10 each Mice were given distilled water, silymarin or Vismisco sample doses equivalent to those in the experiment to studying the effect of increasing the liver damage recovery of Vismisco for days: Mice in groups were given distilled water or reagent for days After days of taking the reagent, they were killed to take carotid artery blood to determine the activity of enzymes AST, ALT; and to take out the liver to observe histopathology (macroscopic, microscopic) and MDA 2.4.2.2 Studying anti-chronic hepatitis and antioxidant effects of Vismisco Mice were randomly divided into groups, 11 each The mice were induced to model chronic hepatitis leading to cirrhosis by injection of peritoneal carbon tetrachloride (CCl4) mixed in olive oil at dose of 10 µ/10g, times/week for 19 weeks The reagent or control was taken daily for 19 weeks (at the same time CCl4 was administered) Group (control): Taking distilled water with a volume of 0.2 ml/10g Group (model): Peritoneal injection CCl4 10 µ/10g, times/week Drink distilled water of 0.2 ml/10g Group (positive control): Peritoneal injection CCl4 10 µ/10g, times/week Silymarin oral dose of 140 mg/kg/day Group (Vismisco dose 1): Peritoneal injection CCl4 10 µ/10g, times / week Vismisco dose equivalent to 0.6 g/kg/day Group (Vismisco dose 2): Peritoneal injection CCl4 110 µ/10g, times / week Vismisco dose equivalent to 1.8 g/kg/day - The mice were injected with CCl4 and reagents for 19 weeks In the 20th week, they were assessed the relative weight, slaughtered, and taken blood and liver to assess liver function, hematological indicators and hydroxyprolin content and observe histopathology (macroscopic, microscopic) of their livers 2.4.3.2 Studying the anti- acute inflammatory effect * Evaluating the anti-inflammatory effect on mouse paw edema model with carrageenin - The rats were separated into groups of 10 animals each Group (model): Taking distilled water 10 ml/kg/day Group (positive control): Taking aspirin at a dose of 200 mg/kg Group (Vismisco dose 1): Taking Vismisco dose equivalent to 0.4 g/kg/day (Taking a dose with an equivalent effect on humans, calculated by a factor of 8) Group (Vismisco dose 2): Taking Vismisco dose equivalent to 1.2 g/kg/day - Rats were given distilled water or drugs for days consecutively before being induced inflammation On the 4th day, 30 minutes after taking the drug, inflammation was induced by injecting carrageenin 1% (mixed in saline) 0.05 ml / mouse into the soles of the right hind feet of the mouse The volume of the mice feet (to the ankle joint) was measured with a specialized tool (Plethysmometer) at the times: before causing inflammation, hours, hours, hours after causing inflammation The increase in leg volume of each mouse (V%) was calculated Inhibition of edema reaction was determined (I%) * Evaluating the anti- acute inflammatory effects by the method of causing peritoneal effusion on white rats: - Rats were randomly divided into groups, 10 each Rats were given distilled water, aspirin or Vismisco sample doses equivalent to those in the experiment to evaluate the anti-inflammatory effect on the model of mouse paw edema by carrageenin - Rats were given distilled water or drugs for days consecutively before causing inflammation On the 4th day, 30 minutes after taking the drugs or distilled water, causing peritonitis with 0.05 g carrageenin solution + 1.4 ml formaldehyde in 100 ml of normal saline, with a volume of ml of peritoneal cavity per rats - After 24 hours of causing inflammation, the rats’ abdominal cavities were opened to suck inflammatory fluid The volume was measured and the number of leukocytes/ml of inflammatory exudate was counted to quantify the protein in the inflammatory exudate 2.4.3.3 Anti chronic -inflammatory effects on an experimental granulomatous model with amiante The mice were randomly divided into groups, 12 each Group (model): Taking distilled water 0.2 ml/10g Group (positive control): Taking methylprednisolon dose of mg/kg Group (Vismisco dose 1): Taking Vismisco dose equivalent to 0.6 g/kg/day Group (Vismisco dose 2): Taking Vismisco dose equivalent to 1.8 g/kg/day - Chronic inflammation was caused by inoculating sterile mg amiante fibers (120°C drying for hour) impregnated with 1% carrageenin, on the nape of the neck of each mouse - After transplanting granulomas, the mice were given distilled water or reagents continuously for 10 days On the 11th day, they were killed with chloroform, peeled off fresh grain tumors to weigh Then tumors were dried 56oC for 18 hours The dried granulomas were weighed Then the average granuloma weight (amiante weight excluded) was compared among groups The antiinflammatory effect was calculated by the percentage of tumor weight reduction compared with the model group, quantification of TNF-α in mouse serum by ELISA method 2.5 Data processing The data were statistically processed by the t - test Student method and "pre post" test (Avant - Après) The data were presented as: X ± SD The difference was significant when p < 0.05 CHAPTER 3: RESEARCH FINIDINGS 3.1 Studying acute toxicity and subchronic toxicity 3.1.1 Acute toxicity Table 3.1 Correlation between Vismisco dose and mice mortality Groups Dose of Vismisco Percentage of Other abnormal (n = 10) (g/kg dead (%) findings Group 144 None Group 216 None Group 288 None Group 360 None In all-male testing animals, signs of neither toxicity nor death among the mice were observed within the critical 72 hours post-administration and to the end of the seventh day Hence the LD 50 of Vismisco evaluated by the Litchfield Wilcoxon method could not be determined (orally) 11 reduced AST and ALT activity compared to group (p < 0.001), equivalent to the biological control group (p > 0.05) 12 Table 3.7 Effects of Vismisco on the activity of MDA in mice poisoned by PAR MDA (nmol/g liver) MDA (nmol/g liver) Groups (2 days after) (4 days after) Group (control) 7.64 ± 1.01 7.56 ± 0.26 Group (model) 9.94 ± 0.86*** 10.67 ± 1.83*** Group (silymarin) 8.27 ± 1.25Δ 8.96 ± 1.12Δ Group Vismisco 0.6g/kg 8.99 ± 1.08 9.53 ± 0.87 Δ Group Vismisco 1.8g/kg 7.79 ± 1.49 8.91 ± 0.33 Δ Comment: The results in table 3.7 showed that MDA in the model group (poisoned by high PAR dose) increased significantly compared to the biological control group (p 0,05) - All of the groups taking Vissmisco and silymarin significantly reduced total bilirubin concentration in mice blood compared to the model group (p < 0.001) * Effects of Vismisco on the degree of coagulation index and hemostasis Tables 3.10 Effects of Vissmisco on the prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen Prothrombin APTT Groups (n = 11) Fibrinogen (g/L) (s) (s) Group (control) 12.54 ± 0.32 22.20 ± 0.48 1.52 ± 0.07 Group (model) 13.54 ± 0.39*** 23.07 ± 0.73** 0.95 ± 0.06*** Group (silymarin) 12.53 ± 0.21 22.22 ± 0.46ΔΔ 1.42 ± 0.06ΔΔΔ Group Vismisco 0.6g/kg 12.32 ± 0.42 21.94 ± 0.58ΔΔ 1.47 ± 0.12ΔΔΔ Group Vismisco 1.8g/kg 12.45 ± 0.24 21.74 ± 0.37ΔΔΔ 1.38 ± 0.10ΔΔΔ Note: ** p < 0,01; *** p < 0,001, p compared with group (control) ΔΔ p < 0,01; ΔΔΔ p < 0,001, p compared with group (model) 14 The results in Table 3.10 show that: - In the model group: Prothrombin time, activated partial thromboplastin time (APTT) were significantly prolonged with statistical significance compared to the control group (p < 0.01; p < 0.001) and the fibrinogen content decreased significantly compared with the control group (p < 0.001) - Oral administration of Silymarin and Vismisco at both doses had no effect on prothrombin time, activated partial thromboplastin time (APTT) and fibrinogen content compared with controls - The oral group of Silymarin and Vismisco at both doses reduced activated partial thromboplastin time (APTT) and increased fibrinogen content compared with the model batch (p < 0.01; p < 0.001) * Effects of Vismisco on indicators assessing the degree of anti-chronic hepatitis Effects of Vismisco on hydroxyproline content in mice liver Tables 3.11 Effects of Vismisco on hydroxyproline content in mice liver Hydroxyproline content Groups (n = 11) (µg Hyp/1g mơ tươi) Group (control) 221.95 ± 8.82 Group (model) 340.26 ± 37.45*** Group (silymarin) 193.47 ± 13.05ΔΔΔ Group Vismisco 0.6g/kg 220.54 ± 29.02 ΔΔΔ Group Vismisco 1.8g/kg 205.14 ± 11.41ΔΔΔ Note: *** p < 0,001, p compared with group (control) ΔΔΔ p < 0,001, p compared with group (model) Comment: The groups using reagents and standard drugs tended to reduce hydroxyproline concentration compared to the model group, but the difference was not statistically significant (p> 0.05) Effects of Vismisco on histopathology in mice liver: Chart 3.1 Effects of Vismisco on histopathology in mice liver 15 The chart above shows that the batch of models has the degree of liver fibrosis on microscopic images with 67% of the samples showing bridging fibrosis (portal-portal space or portal-central space) (3 points) and 33% of patient samples showed fibrosis in the wide portal space (1 point) Silymarin well improved the degree of liver fibrosis on microscopic images with 33% of patients without fibrosis (0 points) and 67% of samples with wide portal fibrosis (1 point) Vismisco significantly improved the degree of liver fibrosis on microscopic images, specifically, 67% of the samples in both groups of Vismisco had no liver fibrosis (0 points) in 33% of the samples Wide portal fibrosis (1 point) * Effects of Vismisco on indicators assessing the degree of antioxidant Table 3.12 Effects of Vismisco on the activity of MDA in mice poisoned by CCl4 Groups (n = 11) MDA (nmol/g liver) Group (control) 8.98 ± 0.59 Group (model) 10.27 ± 1.05*** Group (silymarin) 9.10 ± 0.75 ΔΔΔ Group Vismisco 0.6g/kg 9.43 ± 0.66 ΔΔΔ Group Vismisco 1.8g/kg 8.73 ± 0.69 ΔΔΔ Note: *** p < 0,00;1 p compared with group (control) ΔΔΔ p < 0,001, p compared with group (model) The results in Table 3.12 show that: - The concentration of MDA in the CCl 4-toxic model was significantly higher than that of the biological control (p < 0.001) - MDA levels in mice taking Silymarin and taking Vismisco at both doses were significantly lower with statistical significance compared to the model group (p < 0.001) Table 3.13 Effects of Vismisco on the change of SOD, GPx activities in mouse serum in a model of chronic hepatitis by CCl4 Groups (n = 11) SOD (UI/l) GPx (UI/l) Group (control) 107.07 ± 4.26 9653.34 ± 136.50 Group (model) 21.22 ± 1.56*** 778.25 ± 87.88*** ΔΔΔ 196.86 ± 8.11 5935.66 ± 308.46ΔΔΔ Group (silymarin) 103.38 ± 3.88ΔΔΔ 2838.34± 243.38ΔΔΔ Group Vismisco 0.6g/kg Group Vismisco 1.8g/kg 87.54 ± 3.07ΔΔΔ 1327.65 ± 129,95ΔΔΔ *** Note: p < 0,00;1 p compared with group (control) ΔΔΔ p < 0,001, p compared with group (model) The results in Table 3.13 show that: - The erythrocyte SOD and GPx activities of the model mice significantly decreased compared to the control group (p < 0.001) - Vismisco oral dose of 0.6 g/kg/day had the same SOD activity as the control group and higher than the model batch The erythrocyte GPx activity of mice in this batch was higher than that of the model batch (p < 0.001) 16 - The group taking Vismisco dose of 1.8g/kg/day had higher erythrocyte SOD and GPx activity compared to the model batch (p < 0.001) 3.2.3 Evaluating the anti- acute inflammatory and chronic anti-inflammatory effects of Vismisco 3.2.3.1 Evaluating the anti- acute inflammatory effects of Vismisco * The anti- acute inflammatory effects on the model of carrageenan-induced sewer- rat paw edema Table 3.14 Acute anti-inflammatory effects of Vismisco on the model of carrageenan-induced sewer- rat paw edema After h (V2) After h (V4) After h (V4) % % % Study Degree Reduction Degree Reduction Degree Reduction group of in edema of in edema of in edema /Dose edema compared edema compared edema compared (n = 10) (%) with (%) with (%) with controls controls controls Group 1: 58.66 ± 61.61 ± 67.00 ± water 14.21 15.22 19.01 Group 2: 32.00 ± 52.00 ± 58.00 ± ↓ ↓ 10.19 Aspirin ↓ 45.28 15.60 12.84 16.24 13.1 200 mg/kg ** 66.00 ± Group 3: 66.00 ± ↑ ↑ 68.00 ± ↑ 16.19 15.47 Vismisco 11.86 7.8 17.98 2.06 + 0.4g /kg ++ 55.00 ± Group 4: 53.00 ± 60.00 ± ↓ ↓ ↓ 13.15 13.93 Vismisco 9.36 8.87 2,52 17.59 ++∆ ∆ 1.2g/kg - At hours after causing the inflammation: In a group taken Aspirin, it reduced the apparent volume of rat paw, in a group taken Vismisco with a dose of 1.2 g / kg had tending to reduce the volume of rat paw, However, the difference was not statistically significant A group taken Vismisco with a dose of 1.2g / kg with the effect of inhibiting the swelling response was 8.87% compared to the control - At hours after causing the inflammation: There was no difference in increased volume of rat paw in groups taking Vismisco, Aspirin compared to the control In a group taking Vismisco with a dose of 1.2 g / kg had a 2.52% reduction in edema response to the control - At hours after causing the inflammation: There was no difference in increased volume of rat paw in groups taken Vismisco, Aspirin as compared to the control In a group taking Vismisco with a dose of 1.2g / kg had the most potent effect of inhibiting the swelling response (17.59%) 17 * On the model of carragenin- induced peritonitis Table 3.15 Impact of Vismisco on the volume of inflammatory exudates, an amount of WBC and levels of protein in the inflammatory exudates Groups (n = 10) Group 1: control Volume of inflammatory exudates(ml) Amount of WBC (G/l) Levels of protein (mg/dl) 4.01  1.19 23.74  9.23 3.66  0.21 Group (Aspirin 2,00  0.62*** 16.96  4.44* 3.45  0.33 200mg/kg) Group (Vismisco 4.73  1.58+++ 11.89  4.56** + 3.97  0.19** ++ 0.4g/kg) Group (Vismisco 4.40  1.42+++ 10.85  3.74***++ 3.78  0.22+ 1.2g/kg) - For the volume of inflammatory exudates: In a group taking Aspirin, it had the effect to reduce the volume of inflammatory exudates; in a group taking Vismisco , It had no the effect on decreasing the volume of inflammatory exudates, but there was no difference as compared to the control .- For the number of WBC in inflammatory exudates: Vismisco significantly reduced the number of white blood cells as compared to the group taking aspirin and the control (p