阅读说明：本技术 一种糖苷类化合物在制备预防或/和治疗肝损伤药物中的应用 (Application of glucoside compound in preparation of medicine for preventing or/and treating liver injury ) 是由 王煜丹 赵京浩 程桂广 袁明龙 于 2021-07-29 设计创作，主要内容包括：本发明公开了一种糖苷类化合物的新用途,即其在制备预防或/和治疗肝损伤药物中的应用,属于医药技术领域；本发明从天然植物红香树的叶子中采用超声辅助提取方法,经硅胶柱层析法分离纯化获得本发明糖苷类化合物；细胞凋亡实验及小鼠动物实验结果显示该化合物能降低APAP造成的肝损伤的程度,改善肝组织病变和肝细胞损伤,对APAP诱导的炎症损伤有明显的预防和治疗作用,减少氧化应激和过度炎症造成的机体和细胞损伤,细胞毒性评价结果显示,在100μg/mL以下对细胞存活率无明显影响,该化合物在低浓度时不会对生物机体及细胞健康造成危害,本发明为传统天然植物红香树提供了新的药用途径。(The invention discloses a new application of a glucoside compound, namely the application of the glucoside compound in preparing a medicament for preventing or/and treating liver injury, belonging to the technical field of medicines; the glucoside compound is obtained by separating and purifying leaves of a natural plant, namely the red cedar by a silica gel column chromatography by adopting an ultrasonic-assisted extraction method; the apoptosis experiment and mouse animal experiment results show that the compound can reduce the degree of liver injury caused by APAP, improve liver tissue lesion and liver cell injury, has obvious prevention and treatment effects on APAP-induced inflammatory injury, reduces organism and cell injury caused by oxidative stress and excessive inflammation, and has no obvious influence on the survival rate of cells under 100 mu g/mL as shown by cytotoxicity evaluation results, and the compound does not cause harm to organism and cell health at low concentration.)

1. The application of the glucoside compound with the structural formula shown as the following formula in preparing the medicine for preventing or/and treating liver injury:

。

Technical Field

The invention relates to an application of a glucoside compound in preparation of a medicine for preventing or/and treating liver injury, and belongs to the technical field of medicines.

Background

Acetaminophen (APAP) is considered to be the most common antipyretic analgesic in clinical treatment dosage, but many cold treatment compound preparations contain APAP, which causes that public unconsciously overuse of APAP when buying cold medicine to treat diseases, the excessive medicine easily causes liver injury, and if the liver injury cannot be reasonably controlled in time, serious diseases such as liver fibrosis, liver cirrhosis, even liver failure and the like can be caused. Currently, APAP is still the major source of clinical drug-induced liver injury, as investigated. Typically, following APAP administration, most APAP will be metabolized to the non-toxic glucuronic acid adduct and sulfated by hepatic phase II metabolic enzymes, and then excreted via the urine and feces. A small fraction (about 5-15%) of APAP is also metabolized by cytochrome P450 enzymes to the active metabolite N-acetyl-P-benzoquinone imine (NAPQI), which can be combined with Glutathione (GSH) in hepatocytes to convert to a non-toxic product. While normally reduced GSH in hepatocytes can neutralize NAPQI, excessive APAP can produce excessive NAPQI, leading to depletion of intracellular and mitochondrial GSH stores, leading to overproduction of Reactive Oxygen Species (ROS), resulting in oxidative stress and mitochondrial dysfunction, leading to hepatocyte necrosis and apoptosis.

A currently clinically common viable method for preventing or treating APAP liver injury is the administration of N-acetyl-L-cysteine (NAC) or vitamin C. However, its therapeutic effect is not satisfactory, and NAC can reduce the hepatotoxicity of APAP by increasing the neutralizing capacity of NAPQI by binding directly to GSH or by increasing GSH synthesis. However, the side effects caused in the NAC treatment process are obvious, and the NAC treatment process has stimulation to the gastrointestinal tract, the central nervous system and the respiratory tract, and is easy to cause adverse reactions such as nausea, vomiting, dizziness, bronchospasm, hemoptysis and the like.

Common antioxidants in natural plants include flavones and lignans. In particular, the flavonoid compound containing polyphenol hydroxyl can play an antioxidant role in vivo by resisting lipid peroxidation, eliminating active free radicals and acting on enzymes, and experiments and clinical practices prove that the flavonoid compound has a stronger liver protection effect. It is used clinically in treating acute and chronic hepatitis, liver cirrhosis and toxic liver damage.

At present, no report on the application of the glycoside compound in drug-induced liver injury protection is found.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a plant derived from a red sage tree (Annesleafragrans Wall.) The new application of the glucoside compound is the application of the glucoside compound with the structural formula shown as the following formula in preparing the medicine for preventing or/and treating liver injury:

；

the medicine for preventing or/and treating liver injury is prepared by taking glucoside compounds as active ingredients, is used for preparing the medicine for treating liver injury, and can be added with one or more auxiliary materials acceptable to pharmaceutical preparations, wherein the auxiliary materials comprise conventional fillers, diluents, adhesives, excipients, absorption promoters, fillers, surfactants, stabilizers and the like in the pharmaceutical field, and flavoring agents, pigments, sweetening agents and the like can be added when necessary; or the composition is compounded with other active ingredients to play a role in synergistically preventing and treating liver injury; can be prepared into various pharmaceutically suitable use dosage forms, such as capsules, pills, powder, tablets, granules, oral liquid, injection and the like.

The glycoside compound Confusoside is prepared by taking leaves of a red cedar as a raw material, naturally drying and crushing the leaves, preparing an ethanol extract by adopting an ultrasonic-assisted extraction method, quantitatively analyzing the ethanol extract with the content of 111.84 +/-9.53 mg/g, and separating and purifying the ethanol extract by using a silica gel column.

The invention is dedicated to the experimental research of the prevention and relief effect of the glucoside compound Confusoside on drug-induced liver injury caused by APAP, and after the glucoside compound Confusoside is administered to mice by intragastric administration, APAP is injected into the abdominal cavity for testing, the liver injury symptom is obviously improved.

The results of cytotoxicity experiments show that the survival rate of cells is not reduced within 100 mu g/mL when the human liver cancer cells (HepG 2 cells) are administrated once by 12.5, 25, 50, 100 and 200 mu g/mL Confusoside; the preparation is administered to mice at 20mg/kg and 60mg/kg Confusoside, and no behavior of the mice such as convulsion, tremor, diarrhea, lethargy, dyspnea and coma is seen; the Confusoside does not cause harm to the health of organisms and cells at low concentration.

The invention researches the glycoside compound Confusoside for preventing and relieving liver injury caused by APAP, and proves that the Confusoside can reduce the degree of liver injury caused by APAP, improve liver tissue lesion and liver cell injury, reduce organ indexes, evaluate indexes of alanine Aminotransferase (ALT), aspartate Aminotransferase (AST) and Lactate Dehydrogenase (LDH) of liver injury, promote oxidation resistance, inhibit inflammatory reaction to inhibit apoptosis and reduce organism and cell injury caused by oxidative stress and excessive inflammation, so the Confusoside can be used as a novel medicine for preventing and treating drug-induced liver injury.

Compared with the prior art, the invention has the following advantages:

1. the invention provides a new medicinal approach for the traditional natural plant of the red cedar;

2. the glycoside compound Confusoside does not cause obvious damage to organisms and cells at a lower concentration, can obviously reduce liver damage, and has high safety;

3. the glucoside compound has simple preparation process and rich content in natural plants, and is suitable for industrial production and market popularization and application.

Drawings

FIG. 1 shows the results of the cytotoxicity test of HepG2 administered with different concentrations of Confusoside compounds;

FIG. 2 shows the results of Control group (normal Control group) in which different concentrations of Confusoside inhibited APAP-induced apoptosis and HepG2 was detected for apoptosis by AV/PI and flow cytometry;

FIG. 3 shows the results of APAP groups (APAP administration) with different concentrations of Confusoside inhibiting APAP-induced apoptosis and HepG2 apoptosis detected by AV/PI and flow cytometry;

FIG. 4 shows the results of the NAC + APAP group (acetylcysteine (NAC, 100. mu.g/mL) in combination with APAP (50. mu.g/mL)) for the inhibition of APAP-induced apoptosis by Confusoside at various concentrations, and detection of HepG2 apoptosis by AV/PI and flow cytometry;

FIG. 5 shows the results of the Cf-L + APAP group with different concentrations of Confusoside inhibiting APAP-induced apoptosis, detecting apoptosis of HepG2 cells by AV/PI and flow cytometry (low concentration of Confusoside (50. mu.g/mL) in combination with APAP (50. mu.g/mL));

FIG. 6 shows the results of the Cf-H + APAP group with different concentrations of Confusoside inhibiting APAP-induced apoptosis, detecting apoptosis of HepG2 cells by AV/PI and flow cytometry (high concentration of Confusoside (100. mu.g/mL) in combination with APAP (50. mu.g/mL));

FIG. 7 shows the statistics of the apoptosis rate of different groups of HepG2 cells with different concentrations of Confusoside inhibiting APAP-induced apoptosis; in the figure, NC is a control group, NAC is an NAC + APAP group, Cf-L is a Cf-L + APAP group, and Cf-H is a Cf-H + APAP group;

the data in figures 2-7 are presented as mean ± mean standard deviation (SEM) (5 per group),^# p<0.01 represents comparison with Control group;^* p<0.05 ，^** p<0.01 represents comparison to the APAP group;

FIG. 8 is a schematic diagram showing the results of morphological observation and H & E staining histopathological observation of the liver of APAP liver injury mice with different concentrations of Confusoside;

FIG. 9 shows the liver organ index results of the experiment of APAP liver injury mice with different concentrations of Confusoside;

FIG. 10 shows the results of the biochemical index ALT in the experiment of mice with APAP liver injury induced by Confusoside at different concentrations;

FIG. 11 shows the AST results of the biochemical indicators in the experiment of mice with APAP liver injury induced by Confusoside with different concentrations;

FIG. 12 shows the results of LDH as biochemical indicators in the experiment of mice with APAP liver injury induced by Confusoside at different concentrations;

the data in FIGS. 8-12 are presented as SEM (6 per group).^# p<0.01 represents comparison with group C;^* p<0.05 ，^** p<0.01 represents comparison to group M; c: normal control group, M: APAP (400 mg/kg) dosing, Y: NAC (60 mg/kg) was administered in combination with APAP (400 mg/kg), Cf-L: confusoside (20 mg/kg) was co-administered with APAP (400 mg/kg) at low concentrations, Cf: high concentration Confusoside (60 mg/kg) was administered in combination with APAP (400 mg/kg);

FIG. 13 is the GSH content results of the test of reducing the oxidative stress injury of APAP administration mice with different concentrations of Confusoside;

FIG. 14 shows the results of the content of superoxide dismutase (SOD) in the test of reducing oxidative stress injury in APAP-administered mice with different concentrations of Confusoside;

FIG. 15 shows the results of the determination of Catalase (CAT) content in the test of reducing oxidative stress injury in APAP-administered mice with different concentrations of Confusoside;

FIG. 16 shows the results of the Malondialdehyde (MDA) content in the experiment of reducing oxidative stress injury in APAP-administered mice with different concentrations of Confusoside;

the data in FIGS. 13-16 are presented as SEM (6 per group).^# p<0.01 represents comparison with group C;^* p<0.05 ，^** p<0.01 represents comparison to group M; c: normal control group, M: APAP (400 mg/kg) dosing, Y: NAC (60 mg/kg) was administered in combination with APAP (400 mg/kg), Cf-L: confusoside (20 mg/kg) was co-administered with APAP (400 mg/kg) at low concentrations, Cf: high concentration Confusoside (60 mg/kg) was administered in combination with APAP (400 mg/kg);

FIG. 17 shows inflammatory factor TNF-αContent results;

FIG. 18 shows the inflammatory factor IL-1 in the experiment of reducing the inflammatory injury of APAP-administered mice with different concentrations of ConfusosideβContent results;

FIG. 19 shows the results of the content of IL-6, an inflammatory factor, in the experiment of reducing the inflammatory injury of APAP-administered mice with different concentrations of Confusoside;

FIG. 20 shows the results of the NO content in the experiment of reducing the inflammatory injury of APAP-administered mice with different concentrations of Confusoside;

the data in figures 17-20 are represented by SEM (6 per group),^# p<0.01 represents comparison with group C;^* p<0.05 ，^** p<0.01 represents comparison to group M; c: normal control group, M: APAP (400 mg/kg) dosing, Y: NAC (60 mg/kg) was administered in combination with APAP (400 mg/kg), Cf-L: confusoside (20 mg/kg) was co-administered with APAP (400 mg/kg) at low concentrations, Cf: high concentrations of Confusoside (60 mg/kg) were administered in combination with APAP (400 mg/kg).

Detailed Description

The invention is further illustrated below with reference to experimental data and the accompanying drawings. These experimental examples are only for illustrating the present invention and are not intended to limit the scope of application of the present invention. After reading the description of the present invention, those skilled in the art will be able to make various changes, modifications and alterations equivalent to the embodiments of the present invention without specific indications, which are made from conventional commercial products or reagents prepared by conventional methods, and without specific indications, which are made by conventional methods.

Example 1: separation and purification of glycoside compound Confusoside

1) Drying leaves of the red fragrant tree collected from Yongde county of Lincang city, Yunnan province of China naturally and crushing;

2) mixing a red scented leaf powder sample with 80% ethanol water solution according to the ratio of material to liquid g: mL of 1:10, performing ultrasonic extraction for 30min, filtering to obtain an extracting solution, repeatedly extracting filter residue for 3 times, mixing the extracting solutions, centrifuging at 4000rpm for 10min, taking supernate, and performing reduced pressure evaporation concentration by using a rotary evaporator to obtain an extractum red scented leaf extract;

3) pre-freezing the extract, freeze-drying at low temperature by a freeze concentration dryer to obtain an ethanol extract of the leaves of the red sage root, and storing in a sample dryer;

4) weighing the ethanol extract of the leaves of the red sage, separating and purifying by using a silica gel column, and performing CHCl₃Eluting with-MeOH (volume ratio of 30: 1), collecting eluent, concentrating, dissolving with 80% methanol water, and crystallizing to obtain Confusoside;

5) the compound structure was identified by Nuclear Magnetic Resonance (NMR) as follows:

Confusoside is yellow amorphous powder; 1H (DMSO-d6, 400 MHz)δ: 12.50 (1H, s, OH-2′), 9.16 (1H, s, OH-4), 7.05 (2H, d, J = 8.3 Hz, H-2/H-6), 6.66 (2H, d, J= 8.3 Hz, H-3/H-5), 6.55 (1H, d, J = 1.8 Hz, H-3′), 6.58 (1H, dd, J = 1.8, 8.9 Hz, H-5′), 7.90 (1H, d, J = 8.9 Hz, H-6′). ¹³C NMR (DMSO-d6, 100 MHz) δ: 130.9 (s, C-1), 129.2 (d, C-2/C-6), 115.0 (d, C-3/C-5), 155.5 (s, C-4), 114.4 (s, C-1′), 163.3 (s, C-2′), 103.4 (d, C-3′), 163.5 (s, C-4′), 108.3 (d, C-5′), 132.6 (d, C-6′), 40.0 (t, C-α), 28.9 (t, C-β), 99.6 (d, C-1″), 73.1 (d, C-2″), 76.4 (d, C-3″), 69.5 (d, C-4″), 77.1 (d, C-5″), 60.5 (t, C-6″), 204.4 (s, CO). HRESIMS （m/z）419.1363 [M - H]^- (calcd. for C₂₁H₂₃O₉, 419.1348)；

。

Example 2: cytotoxicity test of glycoside Compound Confusoside

The cytotoxicity evaluation of the compound Confusoside is carried out by measuring the cell viability by using human liver cancer cells (HepG 2) through an MTT method, and the survival rate of the glycoside compound Confusoside to HepG2 cells is researched by the following specific method:

taking out the frozen HepG2 cells from the liquid nitrogen tank, recovering the cells, culturing for three generations, and carrying out an MTT (methanol to transfer) experiment; HepG2 cells at 3X 10⁴Inoculating the strain/well density to a 96-well plate, incubating for 24h, and grouping experiments after the incubation is finished; blank group (NC): 200μTreating HepG2 cells for 24h by using an L complete culture medium; sample group: 200. mu.L of Confusoside solutions with concentrations of 12.5, 25, 50, 100 and 200. mu.g/mL were selected to treat HepG2 cells for 24 h. After the treatment was completed, the cells were incubated with 0.5mg/mL MTT solution (150. mu.L/well) for 4h, then dissolved in 150. mu.L dimethyl sulfoxide (DMSO), the absorbance of each well was recorded at 570nm using a SpectraMax M5 microplate reader, and the cell viability was calculated by the following formula:

as shown in FIG. 1, the cell viability showed a dose-dependent decrease with the increase of the concentration of the glycoside compound Confusoside, and the cell viability was less than 90% at a concentration of 200. mu.g/mL, which indicates that Confusoside inhibits the proliferation of HepG2 cells at 200. mu.g/mL and shows a certain toxic effect, while Confusoside does not have obvious cytotoxicity at a concentration of 100. mu.g/mL or less.

Example 3: protective action of glycoside compound Confusoside on acetaminophen-induced apoptosis

The effect of the compound Confusoside on APAP-induced apoptosis of HepG2 was examined using Annexin V-FITC (AV)/PI apoptosis kit.

HepG2 cells in 12-well plates (cell number 1.5X 10)⁵One/well) for 12h, when the cells are close to the full-length well plate, the cells are dosed, and the Control group: 1mL of DMEM complete medium was used to treat the cells; APAP group: 1mL of DMEM complete medium was used to treat the cells; NAC + APAP group: 1mL of 100. mu.g/mL NAC-treated cells; Cf-L + APAP group: 1mL of 50. mu.g/mL Confusoside-treated cells; Cf-H + APAP group: 1mL of 100. mu.g/mL Confusoside-treated cells; after 24h, the previous culture medium was discarded, and the cells were subjected to further drug administration treatment, Control group: 1mL of DMEM complete medium was used to treat the cells, and 1mL of 50mM APAP was used to treat the cells for 24h for the remaining groups; after the treatment, the cells were collected in a 1.5mL centrifuge tube, washed 3 times with cold PBS, added with 100 μ L of binding buffer, 5 μ L of Annexin V-FITC and Propidium Iodide (PI), incubated for 10min at room temperature in the dark, and after completion, the apoptosis was detected by flow cytometry.

As shown in FIGS. 2 to 7, the detection of apoptosis by flow cytometry significantly induced apoptosis after APAP treatment compared with Control group (ii) apoptosisp<0.01), but 3 administration groups significantly inhibited APAP-induced apoptosis compared to the APAP group: (p<0.01), and the Confusoside sample set significantly inhibited in a dose-dependent manner.

Example 4: prevention and treatment effects of glycoside compound Confusoside on APAP-induced liver injury of mice

1. Experimental methods

Selecting 30 male Kunming mice with the weight of 20-25 g, adaptively feeding all the mice for 7d before formal experiments, wherein the feeding environment is 24 +/-1 ℃ at room temperature and 50 +/-10% of relative humidity, and performing light and shade alternation every 12 h. Mice were fed with laboratory standard feed and double distilled water. After 7 days of acclimation to the experiment, the mice were randomly divided into 5 groups of 6 mice: group C (control group), group M (APAP model group), group Y (NAC group), group Cf-L (Confusoside low dose group), and group Cf-H (Confusoside high dose group). Normally feeding mice, and respectively carrying out intragastric administration on different groups of mice for 7 days continuously, wherein the dose of each intragastric administration is 0.2 mL;

group C: distilled water;

and M groups: APAP (400 mg/kg) administration;

group Y: 60mg/kg (drug/mouse body weight) NAC (N-acetyl-L-cysteine) was administered in combination with APAP (400 mg/kg);

group Cf-L: 20mg/kg (drug/mouse body weight) of Confusoside was co-administered with APAP (400 mg/kg);

group Cf-H: 60mg/kg (drug/mouse body weight) of Confusoside was co-administered with APAP (400 mg/kg);

after 7 days of continuous administration, mice were fasted overnight, and on the 8 th morning, APAP was dissolved in PBS, and a volume of PBS was injected into mice in group C by intraperitoneal injection, and PBS (400 mg/kg APAP/mouse body weight) containing APAP was injected into mice in group M, group Y, group Cf-L, and group Cf-H.

After 6h, the mice were subjected to eyeball bleeding and euthanized; mice were dissected, liver tissue was observed, collected and used for analysis.

Observing the morphological change of the liver, taking a picture, weighing the liver tissue weight of each mouse, calculating the liver organ index, and randomly preparing H from 2 liver tissues in each group&E staining the sections. Centrifuging mouse blood at 1800 Xg and 4 deg.C for 10min, and collecting blood plasma; ALT, AST, LDH, NO, tumor necrosis factor-α(TNF-α) Interleukin-6 (IL-6) and interleukin-1β(IL-1β) And the biochemical indexes are equal.

Mixing the rest mouse liver tissue with 0.9% sodium chloride solution, and homogenizing with ultrasonic cell crusher; the homogenate was centrifuged at 10000rpm and 4 ℃ for 10min, the supernatant was collected and stored at-80 ℃ for further use; preparing liver homogenate to measure MDA, CAT, SOD, GSH and other biochemical indexes.

The H & E stained sections were observed and histopathological evaluation of mouse livers was performed.

2. Results

(1) Prevention and treatment of liver injury in APAP-dosed mice with Confusoside at various concentrations

As can be seen from FIG. 8, the liver of normal mice in group C was red, and the envelope was smooth, glossy and elastic. Otherwise, morphological changes of mouse liver were observed. The M groups of livers have deepened color, gray yellow spot necrosis, dull liver surface and hardened texture, and obvious spot hyperemia of individual livers can be seen. Compared with the group M, the liver injury of mice in the group Y, the group Cf-L and the group Cf-H is obviously improved, and the liver tissue morphology is close to that of the group C.

The histopathological evaluation was consistent with morphological observations. The hepatic lobule structure of group C is clear, the hepatic cells have no necrotic degeneration, no inflammatory cell infiltration, the cell nucleus is large and clear, the hepatic cords around the central vein are arranged in a radioactive manner, and the hepatic sinus is normal; the peripheral veins of the hepatic lobules of the M mice all have hepatic cell degeneration and necrosis with different degrees, wherein the necrosis is manifested by coagulability, nuclear compaction or lysis and is accompanied by infiltration of inflammatory cells such as neutrophils, monocytes and the like. The cell cord has normal shape, and vacuole degeneration of cells can be seen in the middle zone; compared with the group M, the hepatic lobules of the group Y, the group Cf-L and the group Cf-H are more complete in structure, the hepatic cords tend to be arranged neatly, the hepatic cell morphology is obviously improved, and inflammatory infiltration and degenerative necrotic cells near the central vein are obviously reduced; the liver injury induced by excessive APAP is obviously improved.

As can be seen from FIGS. 9-12, the liver organ index and the biochemical indicators AST, ALT, and LDH of the liver injury condition were visually evaluated, and the water level of the M groups was significantly increased compared to that of the C group (see that the water level of the M group was significantly increased in the above cases: (see that the water level of the M group was significantly increased in the above cases)p<0.01）；Compared with the group M, the indexes of the group Y and the group Cf-L, Cf-H have obviously reduced water average; the Cf-H group is close to the Y group in level, and the effect is more obvious compared with the Cf-L group.

The results show that the compound Confusoside has obvious prevention and treatment effects on APAP-induced liver injury.

(2) Confusoside with different concentrations for alleviating oxidative stress injury of APAP (apop-activated peptide) administration mice

As can be seen from FIGS. 13 to 16, the liver tissues of rats in group M had significantly reduced SOD and CAT activities (as compared with those in group C) ((for example, SOD and CAT activities in liver tissues of rats in group M) ((for example, SOD and CAT activities in liver tissues of rats in group C)p<0.01), the GSH content is also significantly reduced (p<0.01); compared with the group M, the SOD, CAT and GSH activities of the group Y, the group Cf-L and the group Cf-H are all improved, wherein the increase of the group Cf-H is most obvious and is close to the group Y (the group C and the group C are respectively)p<0.01), which shows that Cf can effectively treat liver tissue oxidative damage caused by APAP, and the dosage and the effect are in positive correlation; furthermore, M groups had significantly elevated MDA levels (p<0.01). Comparison with group C suggested lipid peroxidation in liver tissue; Cf-L group reduced MDA levels compared to M group ((M))p<0.05), the Cf-H group has more obvious effect and is close to the Y group (p<0.01); the activities of CAT, SOD, GSH and MDA are important indexes for evaluating the oxidative stress condition in organisms, and the results show that Confusoside has obvious prevention and treatment effects on APAP-induced oxidative stress injury.

(3) Confusoside with different concentrations for alleviating inflammatory injury of APAP (apo-praecox) administration mice

As can be seen from FIGS. 17-20, the inflammatory factor TNF-α、IL-1βThe content of IL-6 and NO is obviously increased (p<0.01). Compared with the group M, the inflammatory factors TNF-α、IL-1βIL-6 and NO contents are all lower than those of group M, and TNF-αThe content is reduced, but the change is not obvious, and the Cf-H group is obviously reduced by (p<0.01) and the group Y, the group Cf-L and the group Cf-H are all significantly reduced compared with the group Mp<0.01), the Cf-L group is not significantly different from the M group (p>0.05). Similarly, IL-1 in group Y and group Cf-HβThe content is obviously lower than that of group M: (p<0.01), the IL-6 content in the Cf-L group is obviously lower than that in the M group (p<0.05), and group Cf-H is close to group Y: (p<0.01). Cf group NOThe content is obviously lower than that of group M: (p<0.01), the NO content of both the Cf-L group and the Cf-H group is significantly lower than that of the M group (p<0.01), the NO content of both the Cf-L group and the Cf-H group is significantly lower than that of the M group (p<0.01), the NO content of the Cf-L group and the Cf-H group is obviously lower than that of the M group (p<0.01)。TNF-α、IL-1βThe activity of IL-6 and NO is a key index for evaluating the level of inflammation in organisms, so that the results show that Confusoside has obvious prevention and treatment effects on APAP-induced inflammatory injury.