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.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 as the Herba Cistanches water extract, shorten the mice relieving constipation time, suppress the absorption of large intestine moisture.
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 therefore easily recurrence and the transfer of postoperative tumor select 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 extracting method 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 extracting method, it is characterized in that, exsiccant Herba Cistanches chylocaulous is clayed into power, add water, reflux, extract, 2 hours, filter, medicinal residues add the water reflux, extract, 2 times again, each reflux, extract, 1 hour, merge 3 times extracting solution, lyophilization instrument dry extraction liquid obtains brown ceramic powder after the drying, be Herba Cistanches extract.
Described powder is 3: 5 with the mass ratio of add water, and medicinal residues are 3: 10 with the mass ratio of add water.
Described 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.
The specific embodiment
The present invention will be further described below in conjunction with the drawings and specific embodiments.
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 is filtered, and medicinal residues add 500g water again, reflux, extract, 2 times, the each extraction 1 hour merges 3 times extracting solution, lyophilization instrument dry extraction liquid, obtain brown ceramic powder after the drying, be Herba Cistanches extract, it be mixed with the aqueous solution of 300ug/ml with distilled water, standby.
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﹠amp; E dyeing, microscopically tissues observed canceration type and number.
Tissue, form and infiltration degree according to canceration are divided into 3 classes: 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: * represents significant difference P<0.05
The influence that table 2 Herba Cistanches extract is formed splenocyte
Annotate: * represents 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 by 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, may 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, dead cell number can 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.Collect medicine feed group and control group mice feces 5 times at interval in the experimentation, use 16SrRNA oligonucleotide probe in situ hybridization method, detect 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, extract concentrations when 200ug/ml~400ug/ml without any stimulating or inhibitory action.The extract concentrations of mice of feeding in this experiment is a safe concentration.