Application of fructus schizandrae extract in medicine for rescuing paraquat poisoning

Technical Field

The present invention relates to rescue paraguat schizandrae extract in the preparation of the application of the medicament.

Background Art

Paraquat (1, 1 '-dimethyl -4, 4' -bipyridine is made up; paraquat, PQ) is a highly toxic organic contact herbicide, acute parataf poisoning mortality exceeding > 50%^[1], with literature^[2] noted that 30 mg/kg parataf may be fatal, sales market this is equivalent to 20% paraquat solution 7-8ml, the lung tissue into a human body, the active absorbing parataf cause acute lung injury and interstitial pulmonary fibrosis. Paraquat also affect the heart, kidney, adrenal, central nervous system, liver, skeletal muscle and a zoki, and even lead to multiple organ dysfunction syndrome (Multiple   Organ   Disfunction   Syndrome, MODS)^[3].

Parataf acute poisoning materials are also non-specific drug, the currently used therapeutic drug and may have the therapeutic effect of the drug includes: 1. Immunosuppressants: cyclophosphamide, glucocorticoids; 2. Antioxidant treatment of: reduced glutathione (GSH), vitamin family, acetyl cysteine (NAC), quinone oxidoreductase 2 inhibitor, iraq reaches pulls presents , sodium salicylate (SA); 3. Traditional Chinese medicine for treating: some research found that cucurmin, rhubarb, Xuebijing to acute parataf poisoning lung injury has certain curative effect^[3,4].

Schisandra   chinensis (Turcz.) Baill the Magnoliaceae plant schizandrae Schisandra. Or Orostachyos   Wils   Rehd.et   sphenanthera Schisandra. Ripe fruit of drying. The former often uses "schisandra chinensis", the latter often uses "usage". Autumn when the time is ripe fruit picking, the drying or dried, and removing the stem. Tang, and the like contained in "new herbal" "gomi flesh gan Suan , nuclear hard, have a salty", in the name of a fruit. Schisandra chinensis is divided into the south, north two. The ancient medical [...] , the intestines, and, at the earliest top grade the book on Chinese medicine passes through Shennong listed in traditional Chinese medicine, strengthening force of nourishing, medicinal value is extremely high, a health-care effect. Efficacy can be used for the treatment of: collects the lung , kidney, body fluid, separates, arresting seminal emission. Cough with dyspnea treating lung deficiency, xerostomia does thirstily , spontaneous, night sweats, emaciated to adsorb, nocturnal emission spermatorrhoea, process. Modern traditional Chinese medicine pharmacology study found schizandrae extract containing lignan, crude polysaccharide, benzine components and flavonid, and some of the components have been proved to have very good oxidation resistance, removing the oxygen radical, the role of enhancing immunity. Schisandra chinensis phenol needlestirs up the line damage to mitochondrial oxidative stress plays a prominent role in protecting^[5]. The focus of this invention is as a research object to schizandrae extract, PQ poisoning to the mouse study the acute lung injury and the treatment of pulmonary fibrosis.

Content of the invention

The purpose of this invention is to overcome the disadvantages in the prior art, provide schizandrae extract in preparing rescue parataf poisoning the application of the medicament.

2nd purpose of this invention is to provide formulations containing Chinese magnoliavine fruit extract in the preparation of rescue parataf poisoning the application of the medicament.

The technical scheme of the present invention are summarized as follows:

Schisandra chinensis extract in the preparation of rescue parataf poisoning the application of the medicament.

Schisandra chinensis (Schisandraceae) or the usage of the states the fruit of Chinese magnoliavine (Kadsurajaponica).

The preparation containing Chinese magnoliavine fruit extract in the preparation of rescue parataf poisoning the application of the medicament.

The formulation is preferably a tablet, capsule, granules or injection.

Experiment proves that: schizandrae extract has good oxidation resistance, the protective function of the lung, lung injury to paraquat lead to having the remarkable role of protection, the role of the same mechanism may and reduce the oxygen free radicals form, reducing oxidation stress of inflammatory reaction, inhibit TGF-β 1, IL -6 the increase.

Description of drawings

Figure 1 is the large dose paraguat schizandrae extract of mouse survival number after being poisoned.

Figure 2 the large dose paraguat schizandrae extract after being poisoned the food is heavy mouse on average.

Figure 3 is the paraquat poisoning schizandrae extract HE dyeing effect on mouse lung tissue (magnification 100 times and 200 times). A: blank control group B: hundred dahuricus dry group (300 mg/kg) C: paraguat D C group vitamin + : parataf + schizandrae extract group (200 mg/kg) E: parataf + schizandrae extract group (400 mg/kg) F: parataf + schizandrae extract group of (800 mg/kg).

Figure 4 schizandrae extract to paraquat poisoning effect on mouse lung tissue dyeing Masson (magnification 200 times). A: blank control group B: hundred dahuricus dry group (300 mg/kg) C: paraguat D C group vitamin + : parataf + schizandrae extract group (200 mg/kg) E: parataf + schizandrae extract group (400 mg/kg) F: parataf + schizandrae extract group of (800 mg/kg)

Figure 5 the PQ poisoning schizandrae extract of mouse serum GSH and lung tissue of the impact on the level of MDA

Figure 6 schizandrae extract the lung tissue of MICE TGF-β 1 PQ poisoning and IL-6 of the impact on the level of

Mode of execution

Used in the present invention the extract of fructus Schisandrae chinensis ethanol reflux extraction method.

The following discloses a preparation step is to enable those of skill in the art can better xie Ben invention, but not to make any limitation of the invention.

Embodiment 1

Schisandra chinensis extract extraction:

Schisandra chinensis raw material north takes 500g plus 8 times the volume of the raw material quality a concentration of 95% aqueous solution of ethanol of the, heating reflux extraction 2 times, 3h/time, filtration, filtrate is combined, decompression recovering ethanol concentration to extract, lyophilized 48h, shall be red-brown powder, calculated to yield 23.5%, powder treating chest obstruction content of 5.5%.

Other parameters of the extracted, to yield 10%-40% in the range of, powder content is treating chest obstruction 5-20% the extract may be used for the invention.

The extracted schizandrae according to the above-mentioned method, the extraction rate is 10%-40% in the range of, powder content is treating chest obstruction 5-20% the extract may be used for the invention.

Embodiment 2

Pharmacological experiment:

Schisandra chinensis extract the large dose paraguat rescue study of the role of poisoning:

1 material and method

1.1 experimental animals and material

Schisandra chinensis extract (embodiment 1 to obtain), with physiological saline to configuration;

Vitamin C injection: tianjin his Company limited (quasi-word dicom H12020392);

4 age male kunming white level SPF 72 only (Beijing uygur passes advantage China experimental animal technology limited), weight (21 ±2 . 4) g;

20% paraquat liquid: Shanghai shanghai association biological medicine (siayi) ltd. (product standard number GB   19308-2003);

GSH detection kit: nankin jiancheng biological engineering research Institute;

MDA detection kit: blue clouds day ;

Mouse TGF-β 1 and IL-6 (ELISA) kit: the United States Company & D R;

IL-17 antibody: shipment numbers ab79056, abcam Company of the United States.

1.2 method

1.2.1 acute PQ poisoning establishment of lung injury model experiment is divided into:

Blank control group (Con);

paraquat group (PQ);

PQ+SCEs (200 mg/kg, 400 mg/kg, 800 mg/kg) group ; (SCEs is the acronym of the Chinese magnoliavine fruit extract)

 C PQ+vitamin (100 mg/kg) group.

Each group of 12 only.

The establishment of the model: mouse disposable intragastrically PQ liquid, the dosage is 300 mg/kg.

Administration method: in model building PQ+SCEs set after the 24-hour intragastrically SCEs one-time, in model building PQ+vitamin   C set after the 24-hour intraperitoneal vitamin a   C, and PQ Con group of dosage such as water instead of the physiological saline, then each group of MICE daily to the medicine according to the above-mentioned method a, continuous 21 days.

1.2.2 preparation of mouse orbitalis RAT serum, blood naturally solidified at room temperature 20 minutes, centrifugal 20 minutes (3000 R/min). Collecting supernatants, -80 the preservation [...].

1.2.3 preparation of the homogenate of RAT lung tissue after the tissue homogenate detectors, 1600g centrifugal 10 minutes, taking for subsequent determination.

1.2.4 mouse lung tissue dyeing and HE removal of animal lung after dyeing Masson, after observing the general situation, with 10% neutral formaldehyde fixed, embedding, slice. Dyeing HE: the routine haematin and eosin staining 10 minutes; dyeing Masson: dyeing duplicate red liquorponceau acidity 10 minutes, in order to 2% glacial acetic acid aqueous solution for washing, 1% molybdenum acid aqueous solution of differentiation 5 minutes, without washing directly using aniline Aizen 5 minutes, observation under the lens pathological morphology of lung tissue.

1.2.5 serum GSH activity detection detecting MDA and lungand lungtakes the blood serum the organization of the tissue homogenate, that strictly in accordance with the kit for measuring absorbance of the method, according to the formula and MDA is converted into the concentration of GSH.

1.2.6 alveolocapillary TGF-β 1 and IL-6 detection takes the lung tissue homogenate, ELISA kit detecting TGF-β 1 and IL-6, a specific method in accordance with the specification.

1.2.7 immunofluoresce detect lung tissue IL-17 the embedding of the expression level of lung tissue for cryosections (the 6  m), paraformaldehyde fixed, dehydration; IL-17 for antibody 4 the overnight incubation [...] , incubation two anti-, exact, fluorescent microscope observation after the sealing.

1.3 all data statistical method using SPSS   21.0 software for analysis, the data in Expressed, compared between groups using single factor analysis of variance, standard α=0.05 test, P < 0.05 is statistically significant difference.

2 results

2.1 mouse survival and observation of food intake: paraquat group toxicantexposure 1st day of the event of death, survival only in 8th day 1 only animal; PQ+SCEs are significantly improved the survival rate of each group, in particular high-concentration group of (800 mg/kg) animal survival 5 only, and significantly higher feeding paraquat group , schizandrae extract can be rescued by high-concentration acute parataf poisoning (Figure 1, 2).

2.2 observation lens dyeing HE:PQ group of mouse lung tissue compared with the loss of the normal structure Con group, alveoloar walls collapse, alveoloar endourethral with exudate, telangiectasias, Hyperemia performance; PQ+SCEs (200,400,800 mg/kg) group of mouse lung injury pathological change notably reduced compared with the PQ group, and with the increase of the density of the SCEs, the pathologic change is gradually reduced; PQ+SCEs (800 mg/kg) group of mouse alveoloar wall to collapse PQ+vitamin   C group is obviously smaller than that (Figure 3). Dyeing Masson: PQ group of mouse lung tissue with Con performance under the lens compared alveoloar interstitia widness, extracellular matrix also obviously hyperplasia, a large amount of dyed green collagen fiber accretionary deposition, forming extensive fibrosis; and PQ+SCEs group of mouse lung fibrosis degree compared significantly reduced with the PQ; PQ+SCEs (800 mg/kg) group of mouse lung tissue collagen fiber deposition less than PQ+vitamin   C group (Figure 4).

2.2 the   SCEs PQ poisoning mouse serum GSH and lung tissue content of MDA PQ group of MICE serum compared with the content of GSH Con group (p < 0.01) significantly reduced, is markedly elevated content of MDA (p < 0.001); PQ+SCEs each group of serum GSH content significantly increased compared with the PQ group (p < 0.05); PQ+SCEs (800 mg/kg) group of mouse serum GSH content and a marked increase in the compared   C PQ+vitamin (p < 0.05) (Figure 5A). PQ+SCEs (400 mg/kg, 800 mg/kg) MDA content is reduced notably lung tissue (p < 0.01), PQ+SCEs (200 mg/kg) group of mouse lung tissue compared with the content of MDA with no obvious change group PQ; PQ+SCEs (400 mg/kg, 800 mg/kg) group of mouse lung tissue PQ+vitamin   C group content and MDA compared with (Figure 5B) significantly reduced. The above result can be that the larger the concentration of the mouse is able to effectively reduce SCEs poisoning the organism oxidative stress level PQ, reduced oxygen free radical damage to the body.

2.3 the   SCEs TGF-β 1 PQ poisoning mouse lung tissue and IL-6 content of mouse lung tissue PQ group TGF-β 1 and IL-6 Con group average expression level increased significantly compared with the (p < 0.01); TGF-β 1 PQ+SCEs group of mouse lung tissue (Figure 6A) and IL-6 (Figure 6B) expression level significantly reduced compared to the PQ group (p < 0.05);

PQ+SCEs (800 mg/kg) group of mouse lung tissue IL-6   C PQ+vitamin expression level compared with the group without significant difference (p > 0.05). The above-mentioned results show that can effectively reduce SCEs PQ poisoning mouse lung tissue TGF-β 1 and IL-6 expression.

To sum up, the SCEs mouse lung injury caused by PQ has obvious protective role, the role of the same mechanism may and reduce the oxygen free radicals form, reducing oxidation stress of inflammatory reaction, inhibit TGF-β 1, IL -6 the upward.

Schisandra chinensis extract is made according to the conventional method of tablet, capsule, granules or injection.

References

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