CN115607532A - Application of rhein in preparing medicine for improving immunity of old people - Google Patents
Application of rhein in preparing medicine for improving immunity of old people Download PDFInfo
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Abstract
The invention belongs to the technical field of biological medical treatment, relates to a new application of Rhein (Rhein), and particularly relates to Rhein for improving the CD8 of an elderly organism + Novel use of T cell in virus-specific immune response function. The rhein comprises rhein, pharmaceutically acceptable salts or esters thereof, hydrates thereof or mixtures thereof, can be used as a medicament for clinically improving the antiviral infection specific immunity of the elderly, and provides help for improving the prognosis and outcome of patients; simultaneously strengthen the specificity CD8 of the old to the viral vaccine + The T cell immunoreactivity has wide clinical application value.
Description
Technical Field
The invention belongs to the technical field of biological medical treatment. Relates to the application of rhein in preparing medicine for improving the immunity of old people.
Background
The global population is rapidly aging, and according to the data of the world health organization, the population over 60 years old is growing more rapidly than any other stage population almost in every country, and the population over 60 years old is expected to exceed 20 billion by 2050. The proportion of the population 60 years and older is expected to rise to 22% of the general population in the next 40 years. With age, the immune system of the aging host declines in its response, which we call "immune aging," and this aging includes changes in the innate immune system and in the adaptive immune response. Natural immunity is relatively conserved, but healthy elderly are still often associated with high levels of underlying inflammatory states. The T cell changes in the adaptive immune system are particularly prominent. These changes include a global shift from naive T cells to memory phenotype T cells, resulting in a decrease in the number of naive T cells and an increase in memory phenotype T cells. Antigen-specific T cells themselves also change substantially with age.
Studies have shown that CD8 in the elderly organism + T cells have a reduced function of specific immune responses and evidence suggests that CD8 + Decreased T cell diversity is associated with decreased immune responses of elderly hosts against new infections, including influenza virus. Aged (more than 18 months old) mouse CD8 + Significant reduction in T cell diversity, shift in dominant epitopes, limitation of TCR diversity, and even loss of some dominant epitope response to influenza, such as the MHC class I-restricted epitope NP of nucleoprotein NP 366–374 /D b The loss of primary immune response to NP results in a reduced ability to resist viral infection
Overall, immune aging causes the decrease of acquired immune protective function of the body, the decrease of vaccine response capability, the increase of basic inflammatory state and the enhancement of immune inflammatory pathological action, so the incidence of various infectious diseases (including influenza), autoimmune diseases, tumors and the like is increased. In view of CD8 in specific immune response + The T cells play a key role in eliminating virus infected cells and improving the CD8 of the old organism + The T cell specific immune response function is beneficial to the resistance of the elderly host to various virus infectious diseases.
Rhein (4, 5-dihydroxanthoquinone) belongs to the monoanthracene 1,8 dihydroxy anthraquinone derivatives, is an effective component separated and purified from various traditional Chinese medicines such as rhubarb, polygonum multiflorum, polygonum cuspidatum and the like, and has pharmacological effects of protecting liver, resisting inflammation, resisting oxidation, delaying aging, resisting cancer, resisting pathogenic microorganisms and the like. Based on the current situation of the prior art, the inventor of the application intends to provide a novel medicinal application of Rhein (Rhein), in particular to Rhein for improving the CD8 of the old organism + Novel use of T cell in virus-specific immune response function.
Disclosure of Invention
The invention aims to provide a novel medicinal application of Rhein (Rhein) based on the current situation of the prior art, and particularly relates to Rhein for improving the CD8 of the elderly organism + Novel use of T cell in virus-specific immune response function.
Experiments prove that rhein has remarkable effect on regulating and improving the aging immune system, particularly T cell immune function, and the invention provides a new application of rhein in enhancing the anti-virus specific immune response function of CD8+ T cells of an aged organism.
In particular, the invention provides application of rhein in preparing a medicine for improving the immune function of the old organism.
The rhein of the invention comprises, pharmaceutically acceptable salts or esters thereof, hydrates thereof or mixtures thereof.
In particular, the medicament takes rhein as the only active ingredient.
The immune function of the invention is CD8 + T cell antiviral specific immune response function, in particular CD8 + The number of T cells induced increases and the functional effect is enhanced.
The aged organism refers to a human organism with the age of more than 60 years old or a rodent organism with the age of more than 18 months old.
In one embodiment, the present invention sorts CD8+ T cells from an elderly organism collection [ e.g., spleen from an elderly C57BL/6 mouse at an age of 18 months or older, or Peripheral Blood Mononuclear Cells (PBMCs) from an elderly person at an age of 60 years or older ];
in one embodiment, the sorted aged organisms are CD8 + Placing T cells in vitro cell culture environment, adding rhein with a certain concentration into the culture solution, performing incubation culture treatment for a certain time to obtain senile organism (mouse or human body) CD8 treated in vitro by rhein + A T cell;
in one embodiment, the above-described rhein-treated aged organism (mouse or human) CD8 is treated in vitro + And (5) carrying out cell activity detection on the T cells to ensure that the activity is over 90 percent.
In one embodiment, the above-mentioned aged organism (mouse or human body) CD8 treated in vitro with rhein is used as a treatment agent + T cells are centrifugally washed and then are infused into the body of the aged (a mouse or a human body) by an intravenous injection method; geriatric organism (mouse or human) CD8 without in vitro treatment with retinoic acid + T cells can be infused intravenously as control cells.
In one embodiment, the production of virus-specific CD8 in different treatment groups of organisms is detected after being subjected to viral infection, or after viral vaccination + T cell number and secretion of cellular effector.
In one embodiment, rhein is infused into the elderly body (mouse or human) by direct oral or intravenous infusion; aged organisms (mice or humans) without oral or intravenous rhein administration served as controls;
in one embodiment, the production of virus-specific CD8 in the different treatment groups is detected in vivo after being subjected to viral infection, or after viral vaccination + T cell number and secretion of cellular effector.
The invention proves that the rhein drug treatment group for treating CD8 in the elderly organism through the implementation mode + T cell specific immune response function is significantly enhanced. The experimental result strongly suggests that rhein or pharmaceutically acceptable salt or ester thereof, hydrate thereof or mixture thereof has the effect of enhancing the CD8 of the elderly organism + Function of the T cell anti-viral specific immune response.
All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art, including variations of that technology or substitutions of equivalents as would be apparent to one of ordinary skill in the art. The terms "comprising," "including," "having," "containing," or any other variation thereof as used herein are inclusive or open-ended and do not exclude additional unrecited elements or method steps.
Drawings
FIG. 1: detecting the cytotoxicity after the rhein medicament treats the cells, wherein,
cytotoxicity test of the medicine after rhein treatment of the cells for 24h, 36h and 48h at different concentrations (0, 15, 30, 60 and 120 uM), wherein the cytotoxicity is lower than 10 percent (9.6 percent) after rhein treatment of the cells for 24h at 60uM, and the aged CD8 is treated under the condition + T cells have less influence on the activity of cells.
FIG. 2 is a schematic diagram: control CD45.2 after adoptive transfer of Rhein treatment and no treatment + C57BL/6 geriatric mouse CD8 + T cell to CD45.1 + In C57BL/6 mice, receptor (recipient) CD45.1 was infected with influenza virus + C57BL/6 mice, aged CD45.2 after Rhein treatment transferred to different tissues in recipient mice compared to untreated controls on days 8 and 32 after primary infection (i.e., 4 days after secondary infection) + CD8 + The ratio and number of influenza virus specific tetramer staining positive cells in T cells, A-D, sequentially represent Rhein intervention in Spleen (Spleen), mediastinal Lymph Node (MLN), peripheral Blood (Blood), and Lung (Lung), and CD8 activated by C57BL/6 aged mice (CD 45.2) without dry pretreatment + Production of virus-specific CD8 by T cells + T cell proportion (dot pattern on left, proportion and absolute statistics on right); the number of mice per group was 4. As a result, to
Denotes P<0.05,**P<0.01,***P<0.001 Note): let CD45.2 + CD8 + CD44 high A population of T cells.
FIG. 3: comparison of Rhein Stem prognosis in spleen cells after 8 and 32 days (i.e., 4 days after second infection) of PR 8-infected recipient C57BL/6 mice (CD 45.1) with aged CD45.2 of untreated controls + CD8 + T cells secrete effector factors of Granzyme A, granzyme B, TNF-alpha and IFN-gamma after being stimulated and activated by influenza virus specific polypeptide,
rhein intervention with activated CD8 in spleen of C57BL/6 aged (CD 45.2) mice without dry pretreatment + The T cells secrete cell effector factors of Granzyme A, granzyme B, TNF-alpha and IFN-gamma (A, dot pattern), the proportion statistical chart (B) of the cell effector factors, the absolute number statistical chart (C) of the cells secreting the cell effector factors and the MFI value statistical chart (D) of the cell effector factors, 4 mice in each group are tested, and the result is that the number of the mice in each group is 4
Denotes P<0.05,**P<0.01,***P<0.001 Note): let CD45.2 + CD8 + CD44 high A population of T cells.
FIG. 4 is a schematic view of: PR 8-infected recipient C57BL/6 mice (CD 45.1) day 8 and day 32 (i.e., 4 days post secondary infection), comparison of Rhein Dry prognosis in mediastinal lymph nodes with aged CD45.2 of untreated controls + CD8 + T cells secrete effector factors of Granzyme A, granzyme B, TNF-alpha and IFN-gamma after being stimulated and activated by influenza virus specific polypeptide,
rhein intervention with activated CD8 in mediastinal lymph nodes of C57BL/6 aged mice (CD 45.2) without dry pretreatment + T cells secrete cellular effector factors Granzyme A, granzyme B, TNF-alpha and IFN-gamma proportion (A, dot pattern), cellular effector factor proportion statistical chart (B), cellular absolute number statistical chart (C) of secreted cellular effector factors, MFI value statistical chart (D) of cellular effector factors, and the number of mice in each group is 4. As a result, to
Denotes P<0.05,**P<0.01,***P<0.001 Note: let CD45.2 + CD8 + CD44 high A population of T cells.
FIG. 5 is a schematic view of: PR 8-infected recipient C57BL/6 mice (CD 45.1) day 8 and day 32 (i.e., 4 days after second infection), rhein Dry prognosis was compared in peripheral blood with aged CD45.2 of untreated controls + CD8 + T cells secrete effector factors of Granzyme A, granzyme B, TNF-alpha and IFN-gamma after being stimulated and activated by influenza virus specific polypeptide,
rhein intervention with activated CD8 in peripheral blood of C57BL/6 aged mice (CD 45.2) without dry pretreatment + T cells secrete cellular effector factors of Granzyme A, granzyme B, TNF-alpha and IFN-gamma (A, dot pattern), a ratio statistical chart of the cellular effector factors (B), an absolute number statistical chart of cells secreting the cellular effector factors (C) and an MFI value statistical chart of the cellular effector factors (D), and the number of mice in each group is 4. As a result to
Denotes P<0.05,**P<0.01,***P<0.001 Note): let CD45.2 + CD8 + CD44 high A population of T cells.
FIG. 6: PR 8-infected recipient C57BL/6 mice (CD 45.1) day 8 and day 32 (i.e., 4 days after second infection), comparison of Rhein dry prognosis in lung tissue with aged CD45.2 of untreated controls + CD8 + T cells secrete effector factors of Granzyme A, granzyme B, TNF-alpha and IFN-gamma after being stimulated and activated by influenza virus specific polypeptide,
rhein intervention with activated CD8 in lungs of C57BL/6 aged (CD 45.2) mice not pretreated by dryness + T-cell secretion of cellular effectors Granzyme A, granzyme B, TNF-alpha, IFN-gamma ratio (A, dot pattern), cellular effector ratio histogram (B), cellular absolute count of secreted cellular effectors (C), MFI value histogram (D) of cellular effectors (4 mice per group of experiment)
Denotes P<0.05,**P<0.01,***P<0.001 Note): let CD45.2 + CD8 + CD44 high A population of T cells.
Detailed Description
Example 1C57BL/6 geriatric mice (CD 45.2) + ) Preparation of spleen cells
Taking SPF grade 18-24 month old female C57BL/6 mice (CD 45.2) + ) 12 mice were first sacrificed by cervical dislocation, fixed on a dissecting plate, sprayed on the body surface with 75% sterile alcohol, and the spleen was dissected with an autoclave dissecting instrument.
(1) A10 cm cell culture dish is put into a 200-mesh filter screen with proper size, and then 3ml of R10 culture solution is added into the culture dish. Spleen was placed on filter screen in petri dish and 5ml syringe was ground
(2) After grinding, a 15ml centrifuge tube was taken and marked on the wall. And transferring the cell suspension into a centrifuge tube by using a 1ml pipette, and inserting the centrifuge tube into an ice box after the cell suspension is transferred, so as to ensure the activity of the cells.
(3) The supernatant was discarded after centrifugation at 500g for 5 minutes at 4 ℃. Add 1ml of 1 × schizophyllum solution, mix gently and place on ice for 5 minutes.
(4) 4ml of R10 culture solution is added and mixed evenly, and the red cracking is stopped. The supernatant was discarded after centrifugation at 500g for 5 minutes at 4 ℃. (5) Add 1ml of R10 culture solution to resuspend the cells, aspirate the cell suspension filter with a 1ml pipette gun, and transfer to a new centrifuge tube.
(6) Counting the spleen cells: 10ul of cell suspension was aspirated and the cells were diluted 200-fold for counting.
Example 2C57BL/6Aged mouse (CD 45.2) + ) Spleen CD8 + Sorting of T cells
(1) Centrifuging 500g of spleen cells obtained in the experimental technical scheme 1 for 5min, discarding supernatant and MojoStor TM Buffer resuspended splenocytes (Buffer was ready to use).
(2) Filtering the cells with a 70 μm cell filter, centrifuging for 5min at 300g, and adding MojoCort of appropriate volume TM Buffer to a final concentration of 1x10 8 /ml。
(3) Each 100ul (10) 7 Cells) was added to 10ul of Biotin-antibody cocktail, mixed well and incubated on ice for 15min.
(4) Every 10 th 7 The cells were added with 10ul Streptavidin Nanobeads, mixed well, incubated on ice for 15min
(5) 2.5ml of MojoCoort was added to the cells TM Buffer, gently blow and mix.
(6) The column was placed in a magnetic sorter and 3mL of cell separation buffer was added to the column to rinse the column.
(7) The cell suspension was added to the column and after 5min unlabelled cells of interest were collected.
(8) The cells were transferred to a 15ml centrifuge tube, centrifuged at 300g for 5min, the supernatant was discarded, 1ml of R10 was resuspended, and the cells were counted.
Example 3 working conditions of Rhein-treated cells were investigated
Rhein (stock concentration: 1000 uM) was first diluted with R1640 medium in gradient 0, 15uM, 30uM, 60uM, 120uM, with three duplicate wells set at each gradient. Three treatment times (mouse splenocytes) were set simultaneously: 24h, 36h and 48h. Method for detecting cytotoxicity by using kit LDH-Cytox TM The Assay Kit measures the magnitude of cytotoxicity of the drug at different concentrations and for different periods of time. The conditions of use of the drug were determined as having cytotoxicity lower than 10%.
(1) Preparing a culture medium: RPMI 1640+5% Biantibody +2 mmol/l-glutamine + 50. Mu. Mol/l beta-mercaptoethanol
(2) Three 96-well plates are provided, and are marked according to the time for treating the cells. Aspirate 100ul of cell suspension diluted with medium into 96-well platesInspection hole (2 x 10) 6 Hole/bore). High control, low control, background control were set simultaneously, each control was set with three duplicate wells. 100ul of cell suspension was added to each of the high and low control wells, and 100ul of cell-free medium was added to the background control
(3) 100ul of drug-containing medium was added to each well of the experimental wells and 100ul of drug-free medium was added to the control wells.
(4) Respective 96 well plates at 37 ℃ 5% CO 2 Culturing in an incubator for 24h, 36h and 48h.
(5) After 10ul Lysis Buffer was added to the high control wells, 10ul medium was added to the low and background control wells. 5% CO at 37 ℃ 2 Culturing in an incubator for 30min.
(6) The 96-well plate was centrifuged for 5min (500 Xg) to pellet the suspension cells.
(7) Aspirate 100ul of supernatant from each well into a new 96 well plate.
(8) After adding 100ul of Working Solution to each well, incubation was carried out for 15min at room temperature in the dark.
(9) Immediately after 50ul Stop Solution was added to each well, the absorbance at 490nm was measured using a microplate reader.
(10) Calculation of the magnitude of cytotoxicity
A is the absorbance of the sample; b, absorbance of high control; c absorbance of Low control
EXAMPLE 4 Rhein vs. C57BL/6 aged mice (CD 45.2) + ) Spleen CD8 + Treatment of T cells
(1) Preparing a culture medium: RPMI 1640+5% Biantibody +2 mmol/l-glutamine + 50. Mu. Mol/l beta-mercaptoethanol
(2) Magnetic bead sorted CD8 + The T cells were resuspended in medium and the cell concentration was adjusted to 2X10 7 /ml。
(3) Pipette 100. Mu.l of cell suspension into 96-well plates (2X 10) 6 Per well), 24 wells were plated (12 wells each for experimental and control cells).
(4) Mu.l of Rhein-containing medium (final concentration: 60. Mu.M) was added to each well of the experimental group, and 100. Mu.l of drug-free medium was added to the control group, and gently mixed by pipetting using a pipette gun.
(5) The 96-well plates were incubated at 37 ℃ in a 5% CO2 incubator for 24h.
(6) After the treatment, 500g was centrifuged for 5 minutes, the medium was discarded, washed 2 times with PBS, and then the PBS was resuspended, the cells were collected, and the cells were counted.
Example 5 geriatric mouse CD8 + T cells (CD 45.2) + ) Adoptive transfer to congenic C57BL/6 mice (CD 45.1) + )
(1) 24 SPF-grade 8-12 week-old female C57BL/6 mice were selected and divided into drug experimental groups and control groups of 12 mice each. Note: three days before the start of the experiment we transferred the mice to the biosafety class 2 laboratory in the laboratory animals department. Mice were grouped, 4 per cage, and labeled by earhole punching.
(2) The drug-treated cells and untreated control cells (cell amount: 2X 10) were aspirated by a 1.5ml insulin needle 6 120ul PBS) was injected into CD45.1 separately by retroorbital venous plexus injection + In vivo in mice.
(3) 24h after adoptive, at 0.5LD 50 PR8 influenza virus nasal drops infected recipient mice with 3LD on day 28 post-infection 50 Nasal drip combined 7LD 50 Tail vein injection for the second infection.
Example 6 Rhein-treated group geriatric mouse CD8 + T cell and control group geriatric mouse CD8 + Comparison of T cell response function
In this example, CD8 in the experimental group and the control group on day 8 after infection and day 32 after infection (day 4 after the second infection) were detected mainly by flow cytometry + Viral-specific CD8 induced by influenza infection of T cells + T cells (Tetramer) + CD8 + T) level, and two groups of CD8 stimulated by specific peptide of influenza are detected + The T cells secrete cellular effector factors including the levels of Granzyme A, granzyme B, IFN-gamma, TNF-alpha.
1. Surface dyeing: virus-specific CD8 + Proportion of T cells
(1) Cell plating: adding 5 x10 of the solution into each hole of a 96-hole U-shaped plate 5 -1×10 6 Cells, 4 ℃,500g, centrifuged for 5 minutes and the supernatant discarded.
(2) The cell pellet was shaken up on a shaker, 200. Mu.L of pre-cooled PBS was pipetted to wash the cells, at 4 ℃ and 500g, centrifuged for 5 minutes and the supernatant discarded.
(3) Flow-through chromosome lines were 20 μ L, staining protocol and antibody concentrations used were as follows:
(4) The cell pellet was shaken up on a shaker, 20. Mu.L of the prepared staining mix antibody was added to each well, and gently blown with a pipette gun to bring the antibody into full contact with the cells.
(5) A piece of tin foil paper with proper size is taken to wrap the 96-well plate, protected from light, incubated for 30 minutes at 4 ℃, taken out in the middle, shaken and mixed uniformly for one time.
(6) After staining was complete, 150. Mu.L of staining buffer was added to each well to wash the cells twice, at 4 ℃ and 500g, centrifuged for 5 minutes, and the supernatant was discarded.
(7) Finally, 200. Mu.L of precooled staining buffer resuspended cells were added to each well, transferred to a flow tube and labeled, and then tested on a machine.
2. Cytokine staining
(1) Cell plating: adding 2X10 of the mixture into each hole 6 Individual cell
(2) Influenza dominant epitope peptide segment NP 366-374 Peptides (ASNENMETM) and PA 224-233 Peptides (sslenfranyv) stimulate cells of different tissues. The working concentrations were all 2.5. Mu.g/mL. Furthermore, a Golgi blocker was added at 1ul/ml, and after mixing, the mixture was cultured in a cell culture chamber at 37 ℃ for 5 hours.
(3) After the completion of the culture, the mixture was centrifuged at 500g for 5 minutes at 4 ℃ to discard the supernatant. Cells were washed once with pre-cooled PBS.
(3) Surface staining of cells by the same procedure as described above
(4) 150ul of a membrane rupture solution (Permeabilization solution) was added to each well, gently blown and mixed by a pipette, and the 96-well plate was wrapped with a tin foil paper and protected from light at room temperature for 15 minutes.
(5) After cell perforation, 1000g was centrifuged for 8 min and the supernatant was discarded.
(6) The cell pellet was suspended by vortexer, 150ul 1 × wash buffer was added to each well to wash the cells 2 times, the residual membrane-breaking solution was removed, 1000g was centrifuged for 8 min, the supernatant was discarded, and the cells were suspended by vortexer.
(7) Intracellular cytokine staining
(8) Add 20ul staining mix per well, mix gently, avoid light, incubate for 30 minutes at 4 ℃. Mix well with shaking every 10 minutes.
(9) Add 200ul staining buffer twice per well, wash at 4 ℃ and 1000g, and centrifuge for 8 min.
(10) Cells were shaken up, 200ul staining buffer was added per well, and resuspended cells were transferred to a flow tube and labeled.
(11) And (4) performing flow type computer operation, and analyzing data by Flowjo software.
Example 7 measurement of the magnitude of cytotoxicity after Rhein drug treatment of cells
After rhein respectively treating cells for 36h and 48h, the damage of the medicine to the cells is more than 10%. Cytotoxicity following drug concentration after 24h treatment of cells at different concentrationsIncreased, wherein the cytotoxicity was over 20% when cells were treated with 120uM rhein. Cytotoxicity was less than 10% (9.6%) when cells were treated with 60uM concentration. Thus, elderly CD8 was treated under this condition + T cells had less effect on cell viability (figure 1).
Compared with the control group, the rhein drug treated group of old CD8 + Viral-specific CD8 induced by influenza infection of T cells + The level of T cells is higher. Drug-treated group CD8 in spleen, mediastinal lymph node, lung tissue on days 8 and 32 after primary infection (i.e., 4 days after secondary infection) + T cell-induced virus-specific CD8 + The proportion and the number of the T cells are higher than those of a control group. In peripheral blood, drug-treated group CD8 on day 8 post infection + T cell-induced virus-specific CD8 + The T cell ratio and number were also higher than those in the non-intervention group, and at day 32, the intervention group tended to increase although there was no statistical difference from the non-intervention group (fig. 2).
The drug-treated group had aged CD8 as compared with the control group + T cells secrete higher levels of secreted cellular effector by stimulation with influenza-specific peptides.
In the spleen, drug-treated group activated CD8 + The proportion, the number and the Mean Fluorescence Intensity (MFI) of the Granzyme A secreted by the T cells are obviously increased, the proportion, the number and the MFI of the Granzyme B are obviously increased at the 8 th day after infection, and the proportion, the number and the MFI of the Granzyme B tend to be increased at the 32 nd day; the secretion of TNF-. Alpha.and IFN-. Gamma.were elevated to various degrees in the drug groups (FIG. 3). Drug group activated CD8 in mediastinal lymph nodes + T secretion of Granzyme A, TNF-alpha and Granzyme B were all significantly increased, IFN-gamma was significantly increased at 8 days after infection, and increased tendency was observed at 32 days. Both the absolute number of cellular potency factors and the MFI values increased to different extents (fig. 4). Activated CD8 in peripheral blood + The proportions of the Granzyme A, TNF-alpha and Granzyme B secreted by T are obviously increased, IFN-gamma tends to be increased, and the absolute number of cells and MFI value of each cell effective factor are increased to different degrees (figure 5). In lung tissue, the respective cellular effector factors are present to varying degreesIncreasing trend (fig. 6).
Mean ± sd of all data
The representation was performed and statistical analysis was performed using the statistical mapping software GraphPad Prism7, and statistical comparison of the two sets of data was performed using unpaired t-test statistical analysis. The difference is statistically significant when P is less than or equal to 0.05.
In conclusion, the aged mice treated with rhein had aged organism CD8 + After T cells, their specific immune response function to influenza infection is improved. The main points are as follows: 1) Rhein-dried mouse CD8 + Viral-specific CD8 production by influenza infection of T cells + The number of T cells increases. 2) Rhein treatment of geriatric mouse CD8 + T cells are stimulated with influenza-specific peptides to secrete increased levels of cellular effector.
Claims (6)
1. Application of rhein in preparing medicine for improving immunity of senile organism is provided.
2. Use according to claim 1, characterized in that: the rhein comprises rhein, pharmaceutically acceptable salt or ester thereof, a hydrate thereof or a mixture of the rhein and the salt or the ester.
3. Use according to claim 1 or 2, characterized in that: the rhein in the medicine is used as the only active component.
4. Use according to claim 3, characterized in that: the immune function is CD8 + T cell antiviral specific immune response function.
5. Use according to claim 4, characterized in that: the immune response function refers to CD8 + The number of T cell induction increases and the functional effect increases.
6. Use according to claim 1, characterized in that: an elderly organism refers to a human organism older than 60 years of age, or a rodent organism older than 18 months of age.
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