CN102114079B - A kind of Cistanche extract extraction method and its application - Google Patents
A kind of Cistanche extract extraction method and its application Download PDFInfo
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- CN102114079B CN102114079B CN2011100391162A CN201110039116A CN102114079B CN 102114079 B CN102114079 B CN 102114079B CN 2011100391162 A CN2011100391162 A CN 2011100391162A CN 201110039116 A CN201110039116 A CN 201110039116A CN 102114079 B CN102114079 B CN 102114079B
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Abstract
The invention discloses a method for extracting cistanche deserticola extract and application thereof, belonging to the technical field of medicinal plant application research. The extract is derived from cistanche deserticola or cistanche tubulosa of cistanche genus of Orobanchaceae family. Grinding dry succulent stem of Cistanchis herba into powder, reflux extracting, filtering, reflux extracting residue, and freeze drying to obtain brown powder, i.e. Cistanchis herba extract. The cistanche deserticola extract can activate an immune system, increase the weight of spleen of immune organs, treat intestinal cancer, reduce intestinal hyperplasia, protect normal intestinal epidermal cells and reduce helicobacter pylori infection in the intestinal tract.
Description
Technical field
The invention belongs to medicinal plants applied research technical field, be specifically related to a kind of Herba Cistanches extract, method for distilling and the application aspect preparation treatment bowelcancer medicine thereof.
Background technology
Herba Cistanches has invigorating the liver and kidney, benefiting essence-blood, and the effect of loosening bowel to relieve constipation is returned kidney, large intestine channel.See from Gui Jingshang; Herba Cistanches has the effect of getting twice the result with half the effort to treatment large intestine pathological changes; But the research of modern pharmacology drug effect mainly concentrates on the functions of loosening bowel relieving constipation of Herba Cistanches, and the influence of other large intestine pathological changes is studied seldom, can significantly improve mice intestinal propelling degree like the Herba Cistanches water extract; Shorten the mice relieving constipation time, suppress large intestine absorption of water.
Pharmacological effect article about Herba Cistanches is a lot, but does not all relate to the anti-intestinal cancer effect and the mechanism thereof of Herba Cistanches.Intestinal cancer is the malignant tumor of big epithelium of intestinal mucosa origin, is one of modal malignant tumor of digestive tract, and mortality rate is higher.Excision is the topmost treatment means of intestinal cancer, yet the postoperative tumor is prone to recurrence and transfer, therefore selects hypertrophy, the transfer of rational and effective medicine inhibition tumor cell, and is very useful to improving intestinal cancer patient survival rate.
The animal model of intestinal cancer has a lot, and is wherein comparatively common with TGF-β 1-deficient mice.The Rag2 gene knockout of TGF-β 1-deficient mice (Tgfb1 ± Rag2-/-), and lacking T cell and B cell, caecum and colon position can be inflamed in the mice growth course, cause hyperplasia and canceration in various degree.
Summary of the invention
The object of the invention is to provide a kind of Herba Cistanches extract.
The object of the invention also is to provide the method for distilling of above-mentioned Herba Cistanches extract.
The object of the invention also is to provide the purposes of above-mentioned Herba Cistanches extract.
A kind of Herba Cistanches extract method for distilling is characterized in that, exsiccant Herba Cistanches chylocaulous is clayed into power; Add water, reflux, extract, 2 hours is filtered; Medicinal residues add the water reflux, extract, 2 times again, and each reflux, extract, 1 hour merges 3 times extracting solution; Lyophilization appearance dry extraction liquid obtains brown ceramic powder after the drying, be Herba Cistanches extract.
Said powder is 3: 5 with the mass ratio of add water, and medicinal residues are 3: 10 with the mass ratio of add water.
Said Herba Cistanches extract derives from Desert Herba Cistanches or the Cistanche Tubulosa that the Orobanchaceae Herba Cistanches belongs to.
The application of described Herba Cistanches extract on preparation treatment bowelcancer medicine.
The application of described Herba Cistanches extract on preparation treatment enteritis medicine.
The intestinal health product of described Herba Cistanches extract.
Beneficial effect of the present invention: but Herba Cistanches extract activating immune system of the present invention, the weight of increase immune organ spleen, treatment intestinal cancer reduces the outgrowth generation of intestinal, protects normal intestinal epidermis cell, reduces the intestinal Helicobacter pylori and infects.
Description of drawings
Fig. 1 Western blot detects iNOS and expresses electrophoresis pattern.
Outgrowth number figure in Fig. 2 tissue slice.
Fig. 3 Herba Cistanches extract is protected the bar diagram of normal intestinal epidermis cell.
The bar diagram of 24 hours nitric oxide growing amounts of Fig. 4 RAW264.7 cell.
The specific embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is further specified.
Embodiment 1: the preparation of Herba Cistanches extract
Exsiccant Herba Cistanches chylocaulous is clayed into power, take by weighing this powder 300g, add water 500g, reflux, extract, 2 hours; Filter, medicinal residues add 500g water again, and reflux, extract, 2 times was extracted 1 hour at every turn; Merge 3 times extracting solution, lyophilization appearance dry extraction liquid obtains brown ceramic powder after the drying; Be Herba Cistanches extract, it be mixed with the aqueous solution of 300ug/ml with distilled water, subsequent use.
Embodiment 2: the pharmacological evaluation of Herba Cistanches
With feed intestinal cancer model mice 3 months of the Herba Cistanches extract aqueous solution of 300ug/ml.Experiment finishes the back and observes the spleen variation, gets small intestinal and big intestinal tissue, 10% formalin fixed, paraffin-embedded tissue, H&E dyeing, microscopically tissues observed canceration type and number.
Tissue, form and infiltration degree according to canceration are divided into 3 types: hypertrophy, adenoma, cancer.Outgrowth typical characteristic is that mucosa thickens, the enteraden elongation, but the upper epidermis cell is normal, and normal goblet cell structure is arranged.The flesh layer that is characterized as of adenoma thickens, and goblet cell reduces, and the enteraden of differentiation appears in Submucosa.Intestinal cancer be characterized as the structural deterioration of tangible muscle layer tissue, nuclear atypia appears in and enteraden upper epidermis cellular abnormality.
Embodiment 3: Herba Cistanches extract: the activating immune system experiment
With feed intestinal cancer model mice 3 months of the Herba Cistanches extract aqueous solution of 300ug/ml.Relatively medicine feed group and control group mice immune organ change and separating Morr. cell; Adopt NK cell and macrophage in CD49 and the F4/80 antibody difference labelling splenocyte.
The result shows that the spleen weight of medicine feed group mice is higher than matched group, and splenocyte quantity also is higher than matched group, and there were significant differences (seeing table 1); The NK cell proportion of medicine feed group mice is significantly higher than matched group, and the ratio of macrophage is higher than matched group, but does not have significant difference (seeing table 2).
Table 1 Herba Cistanches extract is to the influence of intestinal cancer model mice spleen organ
Annotate: * representes significant difference P＜0.05
The influence that table 2 Herba Cistanches extract is formed splenocyte
Annotate: * representes significant difference P＜0.05
Embodiment 4: Herba Cistanches extract reduces that intestinal is outgrowth tests
With feed intestinal cancer model mice 3 months of the Herba Cistanches extract aqueous solution of 300ug/ml.Canceration type and number through histologic analysis medicine feed group and the section of matched group intestinal tissue.10 times of visuals field are added up every various canceration types of mice and number down.
The result shows that the hypertrophy number of medicine feed group mice is starkly lower than the matched group (see figure 2), and there were significant differences, the generation of adenoma and intestinal cancer is also had inhibitory action, but do not see significant difference, maybe be less relevant with laboratory animal quantity.
Embodiment 5: Herba Cistanches extract is protected normal intestinal epidermis cell experiment
Select the normal epidermis cell MODE-K of mice cell line for use, with the damage in the cytokine simulation intestinal, the apoptosis ratio of cell behind the FACS detection adding Herba Cistanches extract.
The result shows; Herba Cistanches can be protected normal intestinal epidermal growth (see figure 3); And under mimic impaired environment, it is impaired that Herba Cistanches extract can protect cell to avoid, under the impaired situation of mimic 55% cell; When adding Herba Cistanches extract simultaneously, can dead cell number be reduced to about 35%.This has crucial meaning for cytothesis and the stable state of keeping intestinal.
Embodiment 6: Herba Cistanches extract reduces intestinal Helicobacter pylori infection experiment
Feed 6 week of intestinal cancer model mice with the Herba Cistanches extract aqueous solution of 300ug/ml.The experimentation interval is collected medicine feed group and control group mice feces 5 times, and application 1 6SrRNA oligonucleotide probe in situ hybridization method detects the infection of Helicobacter pylori in the feces.
The result is presented in 5 detections, and 1 subinfection only appears in the medicine feed group, and 3 subinfections (seeing table 3) appear in matched group.Show that Herba Cistanches extract can effectively reduce the infection of Helicobacter pylori in the intestinal, reduce the generation of enteritis.
Table 316SrRNA detects mouse model intestinal Helicobacter pylori and infects
Annotate :+expression feces is infected by Helicobacter pylori;-expression is not infected.
Embodiment 7: Herba Cistanches extract: activate the mouse macrophage experiment
Use mouse macrophage RAW264.7 cell line; Herba Cistanches extract processing with 0ng/mL (contrast), 50 μ g/mL, 100 μ g/mL, 200 μ g/mL was collected supernatant after 24 hours; Positive control is collected supernatant with the Lps processing of 10ng/mL after 24 hours; Detect the generation of this cell nitric oxide (Nitric Oxide), collecting cell detects the expression of nitric oxide synthetase (iNOS) with Western blot.
The result shows that this extract can activating macrophage, and the expression (see figure 1) of inducible nitric oxide synthase promotes nitric oxide to generate (see figure 4).
Embodiment 8: Herba Cistanches extract is to the acute toxicity test of mice
With feed intestinal cancer model mice 3 months of 300ug/ml Herba Cistanches extract aqueous solution.The individual routine of serious side effects do not occur after Herba Cistanches occurring taking, tried the mice well-grown, none death.Cell experiment also shows in addition, and extract concentrations has no when 200ug/ml～400ug/ml to stimulate or inhibitory action.The extract concentrations of mice of feeding in this experiment is a safe concentration.
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| CN101857526A (en) * | 2009-04-13 | 2010-10-13 | 新疆医科大学 | Preparation of galactitol or galactitol extract from Cistanche deserticola and Cistanche deserticola glycosides waste residue liquid and their application |
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| CN101857526A (en) * | 2009-04-13 | 2010-10-13 | 新疆医科大学 | Preparation of galactitol or galactitol extract from Cistanche deserticola and Cistanche deserticola glycosides waste residue liquid and their application |
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Inventor after: Guo Yuhai Inventor after: Jia Yamin Inventor after: Du Caigan Inventor after: Zhu Yanxia Inventor before: Guo Yuhai Inventor before: Jia Yamin Inventor before: Zhu Yanxia |
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