CN112028867B - Tricyclic diterpene compound and extraction method and application thereof - Google Patents

Tricyclic diterpene compound and extraction method and application thereof Download PDF

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CN112028867B
CN112028867B CN202010947134.XA CN202010947134A CN112028867B CN 112028867 B CN112028867 B CN 112028867B CN 202010947134 A CN202010947134 A CN 202010947134A CN 112028867 B CN112028867 B CN 112028867B
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闫炳雄
宋志军
吴云秋
姚彩云
刘喜慧
阮丽君
王硕
周小雷
缪剑华
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Guangxi Botanical Garden of Medicinal Plants
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Abstract

The invention discloses a tricyclic diterpenoid compound and an extraction method and application thereof, wherein the chemical structural formula of the tricyclic diterpenoid compound is as follows:
Figure DDA0002675710610000011
the molecular formula is C 20 H 34 O 3 . The compound can be used for preparing medicaments for treating acute lung injury and medicaments for resisting depression.

Description

Tricyclic diterpene compound and extraction method and application thereof
Technical Field
The invention relates to the technical field of chemical components of strong-drug plant pulp, in particular to a tricyclic diterpenoid compound extracted from strong-drug plant pulp and an extraction method and application thereof.
Background
Zhuang medicine radix Berberidis kunmingensis is a common medicinal material of Guangxi Zhuang nationality, has medicinal overground part, pungent and slightly bitter taste, warm property and fragrant smell, has the effects of dispelling wind and removing dampness, relieving swelling and pain, and clearing and activating the channels and collaterals, and has definite curative effect on treating rheumatic arthralgia, skin itch and other symptoms. Strong drug-plant short pulp is a herbaceous plant which is very easy to plant, but the research and report on chemical components of strong drug-plant short pulp are few at home and abroad, and the research on the aspect of treating lung injury and depression is not reported on the chemical components extracted from strong drug-plant short pulp in the prior art.
Disclosure of Invention
The invention provides a tricyclic diterpene compound aiming at chemical components in strong-drug plant short pulp, and the chemical structural formula of the tricyclic diterpene compound is as follows:
Figure GDA0003690636840000011
the molecular formula is C 20 H 34 O 3 。
2. The method for extracting the tricyclic diterpene compound is characterized in that the tricyclic diterpene compound is the tricyclic diterpene compound according to claim 1, and the extraction method comprises the following steps:
firstly, taking dry strong drug, dwarf shoot stem and leaf medicinal material powder, and leaching the medicinal material powder by using methanol with the volume concentration of 75-95 percent to obtain an extract;
suspending the extract in water, degreasing and extracting by using petroleum ether, extracting an obtained water layer by using ethyl acetate, combining ethyl acetate extracts, and concentrating to obtain thick paste;
and step three, carrying out gradient elution on the thick paste by using petroleum ether and acetone in the volume ratio of 20:1, 8:1, 6:1, 4:1, 3:1, 2:1 and 1:1 in sequence to obtain seven parts, wherein the seven parts are numbered as Fr-A, Fr-B, Fr-C, Fr-D, Fr-E, Fr-F and Fr-G parts respectively, and separating and purifying the Fr-B part by using silica gel and sephadex repeatedly through a preparative chromatography method to obtain a white solid, namely the tricyclic diterpenoid compound.
The invention also provides application of the tricyclic diterpenoid compound in preparation of a medicine for treating acute lung injury.
Preferably, the medicament down-regulates the level of an inflammatory factor, the level of an inflammatory regulator, in a subject with acute lung injury.
Preferably, the inflammatory factors include TNF-alpha, IL-1 beta, IL-6, and the inflammatory regulator includes MPO.
Preferably, the medicament contains a therapeutically effective amount of the tricyclic diterpene compound and a pharmaceutically acceptable carrier.
Preferably, the medicament is formulated into a pharmaceutically acceptable dosage form.
Preferably, the medicament is formulated as an injection.
Preferably, the dosage of the tricyclic diterpenoid compound is not less than 6.6 mg/kg-d.
The invention also provides application of the tricyclic diterpenoid compound in preparation of an antidepressant drug.
Preferably, the agent increases adenylate cyclase activity and increases AC-cAMP signal transduction pathway activity.
Preferably, the medicament is formulated as an oral dosage form.
Preferably, the dosage of the tricyclic diterpenoid compound is not less than 5 mg/kg-d.
The invention at least achieves the following beneficial effects:
1. experiments show that the compound of the invention shows obvious antidepressant activity in a chronic stress rat model and has application in preparing antidepressant drugs.
2. Experiments show that the compound can effectively inhibit increase of white blood cell count in mouse bronchoalveolar caused by LPS, reduce MPO activity in lung tissues, inhibit increase of TNF-alpha, IL-1 beta and IL-6 levels in the bronchoalveolar cells, and obviously increase cAMP content in the lung tissues. Has good research and development values in the aspect of preparing the medicine for treating acute lung injury.
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FIG. 1 is a HR-ESI-MS spectrum of a tricyclic diterpene compound prepared according to example 1 of the present invention;
FIG. 2 shows tricyclic diterpene compounds prepared in example 1 of the present invention 1 H-NMR spectrum;
FIG. 3 shows tricyclic diterpene compounds prepared in example 1 of the present invention 13 C-NMR spectrum;
FIG. 4 shows tricyclic diterpene compounds prepared in example 1 of the present invention 1 H– 1 H COSY spectrum;
FIG. 5 is an HMBC profile of the tricyclic diterpene compound prepared in example 1 of the present invention;
FIG. 6 shows tricyclic diterpene compounds prepared in example 1 of the present invention 13 C DEPT-135 Spectrum;
FIG. 7 is an HSQC spectrum of the tricyclic diterpene compound prepared in example 1 of the present invention;
FIG. 8 is a bar graph of the effect of groups on cAMP content in mice with acute lung injury in experiment 3.
Detailed Description
The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.
It is to be noted that the experimental methods mentioned in the following embodiments are all conventional methods unless otherwise specified, and the reagents and materials are commercially available unless otherwise specified.
< example 1>
Step one, taking 7.0kg of dry strong drug plant dwarf pulp stem and leaf medicinal material powder, leaching for 3 times with methanol with the volume concentration of 75-95% at room temperature for 5 days each time, filtering and combining filtrates, and recovering the methanol to obtain a concentrated extract;
suspending the concentrated extract in water, and degreasing and extracting for 3 times by using petroleum ether and the extract in a volume ratio of 1: 1; extracting the obtained water layer with ethyl acetate and water layer at volume ratio of 1:1 for 3 times, mixing ethyl acetate extracts, and concentrating to obtain 505g soft extract;
and step three, carrying out gradient elution on the thick paste by using petroleum ether and acetone in the volume ratio of 20:1, 8:1, 6:1, 4:1, 3:1, 2:1 and 1:1 in sequence to obtain seven parts, wherein the seven parts are numbered as Fr-A, Fr-B, Fr-C, Fr-D, Fr-E, Fr-F and Fr-G parts respectively, and separating and purifying the Fr-B part by using silica gel and sephadex repeatedly through a preparative chromatography method to obtain a white solid, namely the tricyclic diterpenoid compound.
The chemical structural formula of the tricyclic diterpenoid compound is as follows:
Figure GDA0003690636840000031
tricyclic diterpenes: as a white solid, HR-ESI-MS (see FIG. 1) at m/z 305.2482(calcd for 305.2481) [ m + H-H 2 O]And combining the hydrocarbon spectrum to estimate the molecular formula of the compound to be C 20 H 34 O 3 The unsaturation was calculated to be 4. 1 H-NMR(CDCl 3 600MHz) Table 1 and FIG. 2 show that there are four methyl proton signals in the molecule: delta H 0.73(s,3H, H-19),0.81(s,3H, H-18),1.23(s,3H, H-16),1.11(s,3H, H-17); proton signals on three olefinic carbons: delta H 5.85(1H,dd,J＝17.4,10.9Hz,H-14),δ H 5.24(1H, d J ═ 17.4Hz, H-15) and δ H 5.10(1H, d J ═ 10.9Hz, H-15); one oxygen-substituted methine proton signal: delta. for the preparation of a coating H 3.89(1H, dd, J ═ 9.1,3.3Hz, H-12) and the oxygen substituted methylene signals: delta H 3.07(1H, d, J ═ 10.8Hz, H-20a) and δ H 3.37(1H,d,J＝10.8Hz,H-20β)。 13 C NMR(CDCl 3 150MHz) data (see table 2 and figure 3) showing a total of 20 carbons, 4 methyl groups, 8 methylene groups and 4 methine groups according to DEPT (see figure 6) and HSQC (see figure 7) data, the remaining 4 being quaternary carbons. Wherein the double bond carbon signal is δ C 141.0(C-14) and delta C 113.7(C-15) continuous hydroxyl carbon Signal of delta C 81.6(C-12) hydroxymethyl carbon Signal Δ C 17.2(C-19), the presumed compound is a tricyclic tetramethylThe labdane tricyclic diterpenoid compound has a double bond and two hydroxyl groups (shown in the chemical structural formula).
In the HMBC spectra (see FIG. 5), the methyl proton signal H-16 (. delta.) is seen H 1.23) and C-12 (. delta.) C 8.16)、C-14(δ C 141.0) remote correlation, H-17(δ) H 1.11) and C-7 (. delta.)) C 39.3)、C-9(δ C 60.0) remote correlation, H-18(δ) H 0.81) and C-19 (. delta.)) C 17.0)、C-3(δ C 35.5) and C-5 (. delta.)) C 50.3) remote correlation, H-20 (delta) H 0.72) and C-1 (. delta.)) C 39.2)、C-9(δ C 60.0) remote correlation, it can be determined that the methyl groups are linked at the 13-, 8-, 10-and 4-positions, respectively, and the hydroxymethylene carbon δ C 72.0 to C-4. The position of the double bond may be represented by H-15 (. delta.) H 5.24) and C-13 (. delta.)) C 74.5) and H-16 (. delta.)) H 1.23) and C-12 (. delta.)) C 81.6)、C-14(δ C 141.0) is determined. Bonding of 1 H- 1 The H COSY spectrum (see FIG. 4) shows that H-11 (. delta.) ( H 1.56) and H-12 (. delta.)) H 3.89) are related to COSY and the H-12 and methine carbon signals are delta C 81.6(C-12) is directly related and the other substituted hydroxyl group can be determined to be attached at the 12-position. To this end, delta is transformed C 72.0(C-20；1H,δ H 3.37,d,J＝10.8Hz；1H,δ H 3.07, d, J ═ 10.8 Hz). Therefore, the planar configuration of the compound was determined and named aromatic B. According to the results of single crystal X-ray diffraction experiments, the absolute configuration of the carbon atom at the chiral center of the compound is determined to be 4R,5R,8R,9R,10S,12S and 13S.
Table 1: process for preparing tricyclic diterpenes 1 H-NMR data (600MHz, CDCl) 3 ,δin ppm,J in Hz)
Figure GDA0003690636840000041
Figure GDA0003690636840000051
Table 2: process for preparing tricyclic diterpenes 13 C-NMR data (150MHz, CDCl) 3 ,δin ppm)
Figure GDA0003690636840000052
Figure GDA0003690636840000061
< test 1>
The determination test of the activity of the tricyclic diterpene compound on the adenylate cyclase is as follows:
1. drugs and reagents: the tricyclic diterpene compound extracted in example 1 is abbreviated as the compound in example 1; a cAMP quantitative determination kit; tris (hydroxymethyl) aminomethane (Tris); the other reagents are analytically pure products.
2. The main apparatus is as follows: isolated myocardial perfusion loading (homemade); LS6500 liquid scintillation counter, Beckman corporation, usa.
3. Ex vivo rat cardiac adenylate cyclase activity assay: healthy males, weighing about 370g of rat 1, were decapitated, and then the heart was removed, and the residual blood in the heart was washed with cold Tris-HCl buffer solution at a concentration of 50mmol/L under ice bath conditions. Ventricular tissue 500mg was weighed, 5mL of 50mmol/L Tris-HCl buffer was added, and the ventricular tissue was minced and homogenized. Each 50ul volume of the ventricular homogenate was dispensed into tubes, two tubes were removed and boiled to inactivate as a control, epinephrine (epinephrine group) and 50ul of each compound of example 1 (example 1 group) at different concentrations were added to each tube, and 50mmol/L Tris-HCl containing 2mmol/L MgSO was added to each tube 2 12.5mmol/L theophylline, reacting solution with pH 7.5 to 450ul, adding 50ul of 100mmol/L ATP solution, incubating in 30 ℃ water bath for 5min, boiling for 5min, cooling to room temperature in ice bath, centrifuging at 3000rpm for 10min, taking supernatant 5ml, measuring cAMP with cyclic adenosine monophosphate kit, and subtracting cAMP amount in an inactivation test tube from cAMP amount in a reaction test tube to obtain cAMP amount generated by AC catalysis. The unit of enzyme activity is expressed as pmol cAMP/(mg wet weight 5 min).
4. The detection method comprises the following steps: and detecting the cAMP content in the sample by an ELISA method. The samples collected in the above step were further treated as required by the ELISA kit, and the cAMP content in each sample was measured by ELISA method, and the results are shown in Table 3.
Table 3: measurement of adenylate cyclase Activity
Figure GDA0003690636840000071
P <0.01 compared to control group.
As can be seen from the results in table 3, the compound of the present invention has significant activity of activating Adenylate Cyclase (AC), and increases cyclic adenosine monophosphate (cAMP) concentration in rat heart cells, thereby participating in various cell function regulation, and playing a wide regulatory role in vivo.
< test 2>
The determination test of the antidepressant action of the tricyclic diterpenoid compounds provided by the invention is as follows:
depression is a complex mood disorder disease characterized mainly by depressed mood, decreased speech, mental retardation, and bradykinesia, with a morbidity of about 9% to 18% in western countries. It is currently believed that chronic, low-level stressors are the major contributors to the development and progression of depression. The stress is mainly characterized in that the body is stimulated by internal and external environments to cause dysfunction of hypothalamic-pituitary-adrenal axis (HPA axis) and dysfunction of immune system, thereby causing various behavioral and functional changes. In recent years, it has been found that antidepressant drugs cause an increase in Adenylate Cyclase (AC) activity and an increase in cAMP content, and thus may act through the AC-cAMP signal transduction pathway.
1. Animals, drugs and reagents: male SD rats, 180-200 g. Example 1 extracted tricyclic diterpenes and imipramine (Sigma company, usa). cAMP enzyme immunoassay kit (Sigma, USA); EGTA (purity more than or equal to 97%, Beijing technology, Biotechnology, Inc.); ATP, DTT (seoulman bio, beijing); theophylline (Sigma company, usa); 125I-cAMP radioimmunoassay kit.
2. The instrument comprises the following steps: an ultrasonic homogenate instrument; a 550 model full-automatic enzyme labeling instrument; SN-695B model intelligent exemption 1 measuring instrument.
3. The method comprises the following steps: establishing a chronic stress model of rats, randomly distributing 10 rats in each cage, and feeding the rats under the conditions of room temperature of 21-23 ℃, humidity of 40-60%, natural illumination and free ingestion of drinking water. The chronic stress time course is 20d, once a day, and the time is as follows: 9: 00-14: 00, daily stress stimulation in the following order: performing high-speed horizontal oscillation for 45 min; forced swimming (water temperature 10 ℃): 5 min; braking for 1.5 h; tail clamping (1 cm from tail root): 1 min; water is forbidden for 24 hours; foot sole electric shock (1mA, time course 1s, 1 time per 1 min): 30 min; forced swimming (water temperature 10 ℃): 5 min; fasting for 24 h; braking for 2 h; high-speed horizontal oscillation is carried out for 1 h; tail clipped (1 cm from root): 1 min; water is forbidden for 24 hours; cage changing and isolated culture are carried out for 24 hours; foot sole shock (1mA, time course 1s, 1 time every 1 min): 45 min; forced swimming (water temperature 10 ℃): 5 min; high-speed horizontal oscillation is carried out for 1.5 h; braking for 2.5 h; tail clamping (1 cm from tail root): 1 min; fasting for 24 h; cage changing and isolated culture are carried out for 24 hours. Example 1 groups were: various doses of the compound of example 1 (2.5, 5, 10mg/kg, ig, dissolved in peanut oil) were administered 60min before the chronic stress test each day. Imipramine groups were: imipramine (10mg/kg ip, dissolved in distilled water) was administered 30min before the chronic stress experiment each day. The normal group was: without undergoing chronic stress. The blank control group was: no medications were given daily before chronic stress. At 60min after the administration on day 21 (example 1 group) and 10min after the administration on day 21 (imipramine group), the brains were immediately decapitated and harvested, and the brains were harvested from the normal group and the blank control group, respectively, by decapitation, and the frontal cortex and hippocampus of the rat were separated and stored in liquid nitrogen for later use in the RIA method for AC activity and the ELISA method for cAMP.
4. AC activity by RIA method: taking a proper amount of tissue samples, and carrying out the following steps: 40 Add pre-cooled Tris-HCl buffer (0.05mol/L, pH 7.4) and grind well into a tissue suspension homogenate on an ice bath. An appropriate amount of homogenate was taken and the protein content was determined by the Bradford method. The total reaction volume was 500uL, which contained 50mmol Tris-HCl buffer, 8mmol theophylline, 4mmol DTT, 1mmol EGTA, 1mmol ATP. Adding appropriate amount of tissue homogenate, keeping temperature in water bath at 30 deg.C for 15min, heating in boiling water for 3min to terminate reaction, cooling, centrifuging at 3000r/min for 10min, and collecting supernatant to measure cAMP. The reaction was performed in a double tube, the homogenate was heated in boiling water for 3min in the control tube to inactivate the enzyme, and the other steps were the same as in the experimental tube. cAMP was measured as described in the 125I-cAMP radioimmunoassay kit. The amount of cAMP produced by AC catalysis was subtracted from the amount of cAMP in the reaction tube, and the enzyme activity was calculated from this amount in units of pmol/(mg. min).
4.1, data statistics processing: statistical processing is carried out by adopting SPSS 19 statistical software package, and data are calculated as mean +/-standard deviation
Figure GDA0003690636840000081
Expression, one-way analysis of variance, differences between groups (P)<0.05) Student-Newman-Keuls test was used to calculate the enzyme activity data as shown in Table 4.
Table 4: effect on AC activity in different brain regions of chronically stressed rats (n-10,
Figure GDA0003690636840000082
)
Figure GDA0003690636840000083
Figure GDA0003690636840000091
p <0.01 compared to control.
5. Measurement of cAMP by ELISA: the tissue samples were placed in a stainless steel grinder, liquid nitrogen was added and ground to a fine powder, the tissue was weighed after evaporation of the liquid nitrogen, dissolved in 10-fold volume of 0.1mol/LHCl, centrifuged at 3000r/min at room temperature for 10min, and then diluted with 0.1mol/L HCl supplied for testing. Following the cAMP enzyme immunoassay kit instructions, a standard curve was prepared and the cAMP concentration in the sample was calculated as shown in table 5.
Table 5: measurement results of cAMP Activity in different brain regions of chronically stressed rat
Figure GDA0003690636840000092
P <0.01, P <0.05 compared to control.
The results in tables 4 and 5 show that after 20 days of chronic stress, the cAMP level and AC activity of the frontal cortex and hippocampus of the rat are significantly reduced, and the difference is significant compared with the normal control group, which suggests that the AC-cAMP signal transduction pathway can be damaged by the chronic stress; while the compound of example 1 (5 or 10mg/kg, ig) antagonizes this effect and increases cAMP levels in the corresponding brain regions of rats, the AC activity increases, the group of ig 5mg/kg compound of example 1 achieves the effect of the group of ip 10mg/kg imipramine and the AC-cAMP signaling pathway activity increases. The AC-cAMP signaling pathway plays an important role in the antidepressant effect of the compound of example 1. In conclusion, the compounds of the present invention showed significant antidepressant activity in a chronic stress rat model.
< test 3>
The determination test of the tricyclic diterpenoid compound for treating acute lung injury comprises the following steps:
1. materials: experimental animal ICR mouse, male, body mass 19 ~ 21g, clean grade, experimental environment: the temperature is kept at 22 +/-2 ℃ and the humidity is 60-70 percent, and water and food can be freely drunk.
1.2, reagents and instruments: example 1 extracted tricyclic diterpenes; LPS (lipopolysaccharide); dexamethasone; myeloperoxidase kits; TNF-alpha, IL-1 beta and IL-6ELISA kits; a protein assay kit; cAMP ELISA kit. GF-1 type time-controlled speed-regulating high-speed disperser; ELX800UV type microplate reader; DHG-9145A type electric heating constant temperature air blast drying cabinet.
2. Method of producing a composite material
2.1 model preparation and administration
Mice ALI model was prepared by the method of instilling LPS into the airways, and the mice were randomly divided into a normal saline group, an LPS model group, a dexamethasone group (DXM5mg/kg) and an example 1 group (6.6mg/kg), and 12 mice were administered per group. Mice were anesthetized by intraperitoneal injection of 100g/L chloral hydrate at 280mg/kg, trachea was isolated inactively, LPS (2mg/kg) was added dropwise to the airways of the mice in each group except the saline group, and the saline group was given an equal volume of saline. After 10min, the administration groups (dexamethasone group and example 1 group) were injected with the corresponding dose of drug intraperitoneally at one time, and the normal saline group and the model group were injected with the same volume of normal saline intraperitoneally. Left at 37 ℃ and sacrificed after 6h, the left lung is subjected to bronchoalveolar lavage, and the right lung is collected and stored.
2.2 white blood cell count and Classification in bronchoalveolar lavage fluid
The mice are anesthetized after 6h of modeling, femoral artery bloodletting is killed, right lung is ligated, trachea cannula is exposed, bronchoalveolar lavage is carried out 3 times by 1.5mL of PBS, recovery rate of bronchoalveolar lavage fluid (BALF) reaches 90%, the cells are counted after part of BALF is evenly mixed, total White Blood Cell (WBC) counting is carried out, the rest BALF is centrifuged at 4 ℃ and 250 Xg for 10min, cell sediment smear is taken, Giemsa staining is carried out after air drying, cell classification counting is carried out, including macrophage, neutrophil and lymphocyte, supernatant is taken after centrifugation, and the supernatant is stored at-80 ℃.
2.3 determination of the content of relevant cytokines and proteins in bronchoalveolar lavage fluid
TNF-alpha, IL-6 and IL-1 beta related cytokines in BALF are determined by a double-antibody sandwich ELISA method according to the requirements of a kit specification, and the total protein content in BALF is determined by utilizing a Bio-Rad protein determination kit.
2.4 determination of MPO Activity and cAMP content in Lung tissues
Accurately weighing the lower right lung lobe tissue, preparing 100g/L tissue homogenate by using normal saline according to the requirements of a kit, centrifuging for 10min at 4 ℃ and 12000 Xg, collecting supernatant, and measuring the cAMP content in the tissue homogenate by using a cAMP ELISA kit according to steps, wherein the MPO activity is measured by adopting an enzymology dynamic method.
2.5 statistical treatment
Statistical processing was performed using SPSS 13.0 software. The homogeneity of variance test is carried out on each group of data, and the mean-average comparison of samples adopts the analysis of variance and the Dunnett test.
2.6, the results are shown in table 6, table 7 and fig. 8.
Table 6: influence on the number of leukocytes, the number of neutrophils and the protein content in alveolar lavage fluid of mice with acute lung injury
Group of White blood cell (× 10) 9 /L) Neutrophil (%) Protein content (pmol/L)
Physiological saline group 13.56±3.67 2.85±1.62 281.22±15.42
LPS model group 205.73±68.31 ·· 78.15±11.83 ·· 308.63±14.77 ··
Dexamethasone group 84.68±24.71 ** 54.79±12.65 ** 284.34±14.97 *
EXAMPLE 1 group 70.53±21.22 ** 51.43±11.02 ** 261.32±19.28 *
Comparison with saline group ·· P<0.01; comparison with LPS group * P<0.05， ** P<0.01。
Compared with a normal saline control group, the total number of leucocytes and the percentage of neutrophils in the BALF of the LPS model group are increased (P < 0.01), after the compound of the example 1 is treated, the increase of the total number of leucocytes and the percentage of neutrophils caused by LPS can be reduced (P < 0.01), and the action intensity is equivalent to that of dexamethasone; the protein content in BALF of mice in LPS group is obviously increased (P is less than 0.01) compared with that in normal saline group, and after the compound in example 1 and dexamethasone are given for treatment, the protein content in BALF can be obviously reduced (P is less than 0.05), and the difference has statistical significance.
Table 7: influence on MPO, TNF-alpha, IL-1 beta and IL-6 content in mice with acute lung injury
Figure GDA0003690636840000111
Comparison with saline group ·· P<0.01; comparison with LPS group * P<0.05， ** P<0.01。
The MPO activity in lung tissues of mice in an LPS group is obviously improved (P is less than 0.01) compared with that of a normal saline group, after administration treatment, the MPO activity in a compound group in example 1 is obviously reduced (P is less than 0.05), and dexamethasone is reduced to some extent; compared with normal saline group, the contents of TNF-alpha, IL-1 beta and IL-6 in BALF of LPS group are all obviously increased (P is less than 0.01), the contents of dexamethasone and the like can be obviously reduced, the content of the compound in example 1 can be also obviously reduced (P is less than 0.01), and the effect is better than that of dexamethasone
FIG. 8 is a bar graph of the effect of cAMP levels in mice with acute lung injury in each group (compared to normal saline group) ·· P<0.01; comparison with LPS group ** P<0.01, asterisks on bars indicate statistical significance). From the data in FIG. 8, it was shown that the lung tissue cAMP content was significantly decreased in LPS group compared to normal saline group (P)< 0.01); dexamethasone and the compound of example 1 significantly increased cAMP levels in lung tissue compared to LPS group (P < 0.01).
In conclusion, the data can show that the compound in the example 1 can effectively inhibit the increase of white blood cell count in mouse BALF caused by LPS, reduce MPO activity in lung tissues, inhibit the increase of TNF-alpha, IL-1 beta and IL-6 levels in BALF, and obviously increase cAMP content in lung tissues, and has good research and development values in the aspect of treating acute lung injury.
While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, so that the invention is not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.

Claims (7)

1. The application of the tricyclic diterpenoid compound has the chemical structural formula as follows:
Figure DEST_PATH_IMAGE002
the molecular formula is C 20 H 34 O 3 The tricyclic diterpenoid compound is characterized by being used for preparing a medicine for treating acute lung injury.
2. The use of claim 1, wherein the medicament down-regulates the level of an inflammatory factor, the level of an inflammatory regulator, in a subject with acute lung injury.
3. The use according to claim 2, wherein the inflammatory factor is TNF- α, IL-1 β, IL-6 and the inflammatory regulator is MPO.
4. The use according to any one of claims 1 to 3, wherein the medicament is formulated as an injection and the tricyclic diterpene compound is administered in a dose of not less than 6.6 mg/kg-d.
5. Use according to claim 1, characterized in that the tricyclic diterpenes are used for the preparation of antidepressant medicaments.
6. The use of claim 5, wherein said medicament increases adenylate cyclase activity and increases AC-cAMP signal transduction pathway activity.
7. Use according to claim 5 or 6, wherein the medicament is formulated as an oral preparation and the tricyclic diterpene compound is administered in a dose of not less than 5 mg/kg-d.
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