Umbilical cord mesenchymal stem cells Cocktail treatment hyperglycemia and diabetic nephropathyTechnical field
The present invention relates to hyperglycemia and medicine for treating diabetic nephropathy field, join in particular to umbilical cord mesenchymal stem cellsClose application of a variety of Chinese medicines related to treating diabetes in preparing treatment hyperglycemia and medicine for treating diabetic nephropathy.
Background technology
According to the World Health Organization (WHO), it is expected that the year two thousand thirty, diabetic will be from 200,008,500 in 2010Ten thousand people increase to 409,003,000.Diabetes are also to one of population of China health hazard disease the most serious, diabetesVarious acute and chronic complication be the main reason for causing diabetic dead.Diabetic nephropathy is that diabetic is mainOne of microangiopathies complication, and the main reason for cause End-stage renal failure, how to effectively prevent and treatDiabetic nephropathy is still one of the important topic that diabetes study is faced.
The pathophysiological features of diabetic nephropathy：Microalbuminuria is the earliest clinic of diabetic nephropathy kidney injuryPerformance is formed mainly with kidney in Histological change, such as extrtacellular matrix deposition, glomerular basement membrane thickening and glomerulusMesentery expansion is related.High-grade Proteinuria will occur in the diabetic nephropathy course of disease later stage, and glomerular filtration rate reduces progressive, kidneyHistological change includes glomerulosclerosis, renal tubule interstitial fibrosis and artery hyaloid degeneration.
The therapy of diabetic nephropathy has much at present, including control blood glucose, blood pressure and reduction albuminuria etc..Whole latter stageThe feasible renal replacement therapies of patient, including haemodialysis and kidney transplant, and kidney transplant only carries out B cell at the same timeIn the case of transplanting just effectively, otherwise kidney failure will occur again.Haemodialysis has large effect to the quality of life of patient,Kidney transplant is to treat the most effective means of end-stage renal disease now.However the shortage in kidney source greatly limits kidney transplantIt promotes.Kidney is a kind of organ of terminal differentiation, and regeneration potential is well below other organs of human body.Renal function is not by listA cell executes, but forms a unit by the cell of different function to function, this but also the regeneration of kidney veryIt is difficult.
Mescenchymal stem cell (Mesenchymal stem cells, MSCs) is also referred to as pluripotency mesenchyma stromal cells,It is a kind of non-hematopoietic stem cell, and Tissue distribution is wide, has self-renewing and differentiation capability.MSCs divides first from marrowIt from obtaining, then can also be obtained from its hetero-organization and organ, these tissues and organ include hair follicle, tooth root, brain periosteum, cartilageFilm, corium, bleeding of the umbilicus, umbilical cord, placenta, fat-muscle, lung, liver and spleen.Multinomial research is it has been shown that the MSCs of in vitro culture is specialThe 113 kinds of transcription products and 17 kinds of albumen that opposite sex expression hematopoietic cell is not expressed.According under international cell therapy association (ISCT)Belong to definition people's MSCs minimum standards of mesenchyma and the proposition of tissue stem cell committee member club：A) under Standard culture conditions, MSCsThere must be the adhesion to plastic substrates；B) CD105, CD73, CD90 are positive, CD45, CD34, CD14 or CD11b,CD79a or CD19 and HLA-DR are negative；C) in vitro under the conditions of standard differentiation, it is thin that MSCs can be divided into osteoblast, fatBorn of the same parents and cartilage cell.
Mescenchymal stem cell has many pilot studys in each subject, also has basic research to confirm that kidney cell has again in the recent periodRaw potential, stem-cell therapy injury of kidney, including diabetic nephropathy have some pilot studys.Recently, umbilical cord mesenchyma is dry thinBorn of the same parents (umbilical cord-derived mesenchymal stem cells, UCMSCs) are increasingly by the weight of researcherDepending on.Umbilical cord mesenchymal stem cells have self-renewing and are divided into the ability of different cells.Umbilical cord mesenchymal stem cells derive fromUmbilical cord.Umbilical cord is the waste after delivery of fetus, is derived from a wealth of sources, and ethnics Problem is not present.In addition, depositing due to tire alveolar-capillary barrierIt is substantially reduced and process no pain of drawing materials by the probability of virus pollution in, umbilical cord.More importantly, umbilical cord mesenchymal stem cells connectContinuous repeatedly passage can still keep stem cell properties, be easy to long-term cultivation and can freeze, and proliferative capacity is high, and safely and effectively.People's navelBand mescenchymal stem cell can exist for quite a long time in Mice Body, and single injection does not cause that host's is immune anti-It answers.The characteristic that particular functionality cell is divided into using umbilical cord mesenchymal stem cells is transplanted, and can be substituted irreversible in vivoLoss of function cell, to there is no disease such as tumour, diabetes, blood disease, the cardiac muscle of effective treatment means bad at present at someExtremely, in terms of the treatment of Parkinson's disease, spinal cord injury etc., there is wide potential applicability in clinical practice.It is treated using stem cells technologyDiabetic nephropathy makes great progress in experimental animal, and is likely to become having for future therapeutic diabetesOne of efficacious prescriptions method.
And having righting tonifying Qi effect in traditional Chinese medicine more, modern pharmacy is studies have shown that Radix Astragali, matrimony vine, pueraria lobata, the five tastesThe several kinds of Chinese medicinal materials such as son, Radix Salviae Miltiorrhizae have each active component, their medicinal histories are long, are treatment diabetes in successive dynasties Chinese medicineCommon Chinese medicine type.
The research ideas of China's traditional Chinese medicine prevention diabetes, from the empiricism of traditional clinical experience to more emphasis sectionLearn the medical development of evidence.Include mainly polysaccharide, flavones currently, being found that a large amount of hypoglycemic substances from natural traditional Chinese medicine plantWith alkaloid etc..Alkaloid compound is concerned because of its good hypoglycemic effect, in the past twenty years, largelyPlant alkaloid is extracted separation, and the alkaloid detached at present has thousands of kinds, and majority has hypoglycemic pharmacological activity, answers extensivelyFor medical domain.However, the absorption and metabolic mechanism, active group inhibiting mechanism and stabiliser etc. to its physiological function are gone backLack comprehensively deep understanding.
The western medicine of diabetes is directed to its cause of disease at present, improves insulin resistance, and the guarantor to Pancreatic beta cells functionShield selects the drug that can improve insulin resistance.These drugs are mainly insulin sensitizer, but take these enhanced sensitivities for a long timeAgent can cause patient's oedema and weight gain, only a few patient to there is also serious liver damage.And Chinese medicine preparation then make by pairWith smaller, conducive to being accepted by patients, and meanwhile it is significant in efficacy.
In view of this, special propose the present invention.
Invention content
A kind of umbilical cord mesenchymal stem cells joint plurality of Chinese extract of present invention offer is preparing treatment hyperglycemia and sugarApplication in the sick nephrosis drug of urine.In the present invention, by that will have differentiation capability and between the umbilical cord for treating diabetes potentialityMesenchymal stem cells with hypoglycemic and improvement kidney state and the kidney trouble caused by diabetes have the more of inhibitionKind Chinese medical extract combination is to prepare the drug for treating hyperglycemia and diabetic nephropathy, so as to effectively control blood glucose waterIt is flat, and improve renal function.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme：
Umbilical cord mesenchymal stem cells combine plurality of Chinese extract in preparing treatment hyperglycemia and medicine for treating diabetic nephropathyApplication.
In the present invention, by the way that mescenchymal stem cell and plurality of Chinese extract to be combined, so as to dry using mesenchymaThe differentiation capability of cell is to supplement damaging cells；Simultaneously in the present invention, also can by using plurality of Chinese extractIt is enough further effectively to control blood glucose level, while kidney can also be played a certain protective role, it is effectively improved renal function.
In the present invention, the umbilical cord mesenchymal stem cells are human umbilical cord mesenchymal stem cells.
Optionally, in the present invention, the application still further comprises culture umbilical cord mesenchymal stem cells, then again will cultureThe step of obtained umbilical cord mesenchymal stem cells are with plurality of Chinese extract use in conjunction.
Optionally, in the present invention, the culture is umbilical cord mesenchymal stem cells are primary or freezing and storing umbilical mesenchymal is dryIt after cell recovery, is cultivated in the medium, then carries out secondary culture again.
Optionally, in the present invention, the culture medium is the DMEM/F12 culture mediums containing FBS, penicillin and streptomysin.
Optionally, in the present invention, the content of the FBS is 10%, a concentration of 100U/ml of penicillin and streptomysin.
Optionally, in the present invention, the secondary culture is carried out in the incubator of 5%CO2.
Optionally, in the present invention, the temperature of the secondary culture is 35~38 DEG C；Preferably, in the present invention, the passageThe temperature of culture is 36~37 DEG C.
Optionally, in the present invention, the application still further comprises the umbilical cord mesenchymal stem cells for obtaining secondary cultureAfter digestion, obtain stablizing proliferative cell, and detect, then again with plurality of Chinese extract use in conjunction the step of.
Optionally, in the present invention, the digestion is to be digested using trypsase.
Optionally, in the present invention, a concentration of the 0.20%~0.25% of the trypsase；Preferably, in the present invention, instituteState a concentration of the 0.24%~0.25% of trypsase.
Optionally, in the present invention, the plurality of Chinese extract is Radix Astragali, ginseng, Radix Codonopsis, Radix Salviae Miltiorrhizae, Radix Notoginseng, matrimony vine, Pueraria lobotaOne or more extracts in root, Schisandra chinensis, Rhizoma Atractylodis Macrocephalae, cassia seed, radix rehmanniae recen, sealwort.
Optionally, in the present invention, the plurality of Chinese extract is Radix Astragali, ginseng, Radix Codonopsis, Radix Salviae Miltiorrhizae, Radix Notoginseng, matrimony vine, Pueraria lobotaOne or more extracts in root, Schisandra chinensis, Rhizoma Atractylodis Macrocephalae, cassia seed.
Optionally, in the present invention, which is characterized in that the plurality of Chinese extract is Radix Astragali, ginseng, Radix Salviae Miltiorrhizae, Radix Notoginseng, whiteOne or more extracts in art, Schisandra chinensis.
Optionally, in the present invention, the plurality of Chinese extract be Radix Astragali, ginseng, Radix Salviae Miltiorrhizae, Radix Notoginseng, Rhizoma Atractylodis Macrocephalae, on Schisandra chinensisState arbitrary 6,5,4,3,2 kind of extract.
Optionally, the present invention in, the plurality of Chinese extract be it is one or more in Radix Astragali, ginseng, Radix Salviae Miltiorrhizae three,Add again Radix Notoginseng, Rhizoma Atractylodis Macrocephalae extract.
Optionally, the present invention in, the plurality of Chinese extract be it is one or more in Radix Astragali, ginseng, Radix Salviae Miltiorrhizae three,The extract of Radix Notoginseng, Rhizoma Atractylodis Macrocephalae, Schisandra chinensis is added again.
Optionally, in the present invention, which is characterized in that the plurality of Chinese extract is the extraction of Radix Salviae Miltiorrhizae, Radix Notoginseng, Rhizoma Atractylodis MacrocephalaeObject.
The Chinese medical extract being proliferated suitable for huMSCs and HMC is a concentration of：Radix Astragali 0.1-10mg/ml, Radix Salviae Miltiorrhizae 0.1-10mg/Ml, ginseng 0.1-10mg/ml；Preferably, Radix Astragali, Radix Salviae Miltiorrhizae, ginseng can also select 0.2 respectively, 0.5,0.8,1,1.5,2,3,4,5,7, the addition concentration of 9mg/ml is for cultivating huMSCs or/and HMC proliferation.Above-mentioned Chinese medical extract cultivate reagent can be distinguishedIt can be also used in mixed way using culture cell.Above-mentioned Chinese medical extract cultivate reagent can also be used in mixed way training with following Chinese medical extractSupport cell：Addition Schisandra chinensis 20-150ul/ml or/and Rhizoma Atractylodis Macrocephalae 10-50ul/ml is for cultivating huMSCs proliferation；The preferred five tastesSub also optional 20,30,40,50,60,70,80,90,100,110,120,130,140ul/ml and other similar concentrations are madeFor Chinese medical extract additive capacity, Rhizoma Atractylodis Macrocephalae can be selected 20,30,40ul/ml and other similar concentrations add as Chinese medical extractAdd dosage, is co-cultured with huMSCs or/and HMC.
The amount and concentration that above-mentioned Chinese medical extract is added are obtained using following extracting method：
Ginseng, Radix Salviae Miltiorrhizae, Radix Astragali are taken, is sliced, filtering, drying is ground, and is crossed 80-120 mesh sieve, is obtained fine powder, spare；
Schisandra chinensis, Radix Notoginseng crush, and are put into container, immerse the 6-16 times of alcohol solution dipping for measuring volumetric concentration 50-80%1-10 hours, heating and refluxing extraction 3 times was 1-6 hours each, filtration, merging filtrate, and vacuum distillation removes ethyl alcohol and is concentrated into 60DEG C relative density is that the concentrate of 1.10-1.13 is spare；
The water that Rhizoma Atractylodis Macrocephalae is measured with 12 times, 10 times and 8 times decocts extraction three times respectively, and each 2h merges Aqueous extracts three times, veryIt is spare that sky is concentrated under reduced pressure into 1g medicinal materials/ml extracting solutions.
Compared with prior art, beneficial effects of the present invention are：
In the present invention, by will have differentiation capability and with treatment diabetes potentiality umbilical cord mesenchymal stem cells withWith hypoglycemic and improves kidney state and the kidney trouble caused by diabetes and have the plurality of Chinese extract of inhibitionCombination so as to effectively control blood glucose level, and improves kidney to prepare the drug for treating hyperglycemia and diabetic nephropathyDirty function.
Ten pleasant impression main ingredients are selected from the treatment relevant traditional Chinese medicine prescription of diabetes, its Chinese medical extract is combined, are sievedSelecting can be used cooperatively with umbilical cord mesenchymal stem cells, with hypoglycemic and improve the traditional Chinese medicine extraction composition of kidney state.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show belowThere is attached drawing needed in technology description to be briefly described.
Fig. 1 is the human mesenchymal stem cell cellular morphology under the microscope of different phase and staining conditions, wherein A areThe cellular morphology that human mesenchymal stem cell without induction differentiation processing is arrived in 25 × microscopically observation；B is through adipogenic inductionThe cell dyeing situation that human mesenchymal stem cell is arrived in 10 × microscopically observation after reagent induction differentiation processing；C is through skeletonizationThe cell dyeing situation that human mesenchymal stem cell is arrived in 10 × microscopically observation after induction agent induction differentiation processing.
Fig. 2 is proliferation of the human umbilical cord mesenchymal stem cells (huMSCs) under the conditions of the different Chinese medical extracts of various concentrationFigure.A is Radix Astragali；B is ginseng；C is Radix Salviae Miltiorrhizae；D is Radix Notoginseng；E is Rhizoma Atractylodis Macrocephalae；F is Schisandra chinensis.
Fig. 3 is the vegetative map under the conditions of the different Chinese medical extracts of messangial cell (HMC) various concentration.A is Radix Astragali；bFor ginseng；C is Radix Salviae Miltiorrhizae；D is Radix Notoginseng；E is Rhizoma Atractylodis Macrocephalae；F is Schisandra chinensis.
Fig. 4 is the vegetative map of human umbilical cord mesenchymal stem cells and HMC co-incubations under the conditions of various combination Chinese medical extract.
Fig. 5 is blank control group, human umbilical cord mesenchymal stem cells group and the huMSCs trainings of plurality of Chinese extract mixed processingWhen supporting, ELISA detects supernatant FN and IV collagen content figure.
Specific implementation mode
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art willUnderstand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specificCondition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer isThe conventional products that can be obtained by commercially available purchase.
1 human umbilical cord mesenchymal stem cells culture of embodiment and propagating method
HuMSCs (i.e. human umbilical cord mesenchymal stem cells) is placed on through original cuiture containing 10%FBS, 100U/ml penicillinIn the DMEM/F12 culture mediums of streptomysin；Then, routine culture passes in 37 DEG C, the incubator of 5%CO2；Passage is trainedObtained 0.25% trypsin digestion of cell is supported, and collects stable proliferative cell.
By 106A obtained human umbilical cord mesenchymal stem cells of collecting are suspended in the PBS that 100ml contains 1% bovine serum albumin(BSA)In buffer solution, be used in combination Flow cytometry positive antibody (CD90, CD105, CD73) and negative antibody (CD45, CD34,CD11b, CD19 and HLA-DR) expression.The result is shown in Figure 1, the human umbilical cord mesenchymal stem cells that display culture obtains meet dryCell standard.
The preparation of embodiment 2, Chinese medical extract
Each Chinese medical extract is prepared according to following methods：
Ginseng, Radix Salviae Miltiorrhizae, Radix Astragali are taken, is sliced, filtering, drying is ground, and is crossed 80-120 mesh sieve, is obtained fine powder, spare.
Schisandra chinensis, Radix Notoginseng crush, and are put into container, immerse the 6-16 times of alcohol solution dipping for measuring volumetric concentration 50-80%1-10 hours, heating and refluxing extraction 3 times was 1-6 hours each, filtration, merging filtrate, and vacuum distillation removes ethyl alcohol and is concentrated into 60DEG C relative density is that the concentrate of 1.10-1.13 is spare.
The water that Rhizoma Atractylodis Macrocephalae is measured with 12 times, 10 times and 8 times decocts extraction three times respectively, and each 2h merges Aqueous extracts three times, veryIt is spare that sky is concentrated under reduced pressure into 1g medicinal materials/ml extracting solutions.
The effect that 3 Chinese medical extract of experimental example grows human umbilical cord mesenchymal stem cells and messangial cell
The good huMSCs of growth conditions (human umbilical cord mesenchymal stem cells) and HMC (messangial cell) are chosen respectively with 3×103A/hole is inoculated in 96 orifice plates, and after cell is adherent, the Chinese medical extract co-incubation of various concentration, control group is addedIt is not added with Chinese medical extract, zeroing hole is free of cell, and only plus 100 μ L culture solutions, every group parallel to do 5 multiple holes, and each group culture reaches 4h,For 24 hours, 10 μ LCCK8 solution are respectively added after 48h, 72h, after 37 DEG C are incubated 4h, 490nm wavelength are selected to measure each hole absorbance (OD) value.The effective concentration measured according to cck-8 results.Test result is as shown in Figure 2 and Figure 3.
Radix Astragali, Radix Salviae Miltiorrhizae, ginseng Chinese medical extract known to testing result can effectively facilitate huMSCs and HMC proliferation, andInhibition or toxic effect will not be generated to huMSCs and HMC；And Schisandra chinensis and Rhizoma Atractylodis Macrocephalae to huMSCs and HMC proliferation almost withoutIt influences, Radix Notoginseng but has been surprisingly found that has apparent inhibiting effect to huMSCs and HMC proliferation compared to control group.
The effect that embodiment 4, different Chinese medical extract hybrid combinings grow human umbilical cord mesenchymal stem cells
The final concentration ratio of different Chinese medical extract mixing accordings to the form below is added to by culture and detection method with embodiment 3100 μ L contain in the good huMSCs culture solutions of growth conditions, observe human umbilical cord mesenchymal stem cells upgrowth situation.
The different Chinese medical extract combination matching final concentration ratios of table 1.
As a result such as Fig. 4, Radix Notoginseng inhibits cell growth with obvious effects, and Radix Astragali, Radix Salviae Miltiorrhizae, ginseng are at concentration 0.1-5mg/mlIt is proportionate with vitro growth rates.
Embodiment 5, the foundation of huMSCs and HMC (messangial cell) co-culture system and human umbilical cord mesenchymal stem cells connectionClose Chinese medical extract Experiment on therapy
1xl0 is made in HMC6The cell suspension inoculation of ml, per hole 2ml, waits for cell attachment in 6 orifice plates in 6 well culture platesAnd after replacing culture medium, upper layer cell is put into 6 orifice plates with aseptic nipper.
By the 0.25% pancreas egg enzymic digestion of the human umbilical cord mesenchymal stem cells of secondary culture in embodiment 1, people's umbilical cord is madeMesenchyma stem cell suspension, adjustment cell concentration is 1x106/ml, is inoculated into the set ware soaked in culture solution in advance,It is co-cultured in 37 DEG C of juxtaposition, 5%CO2 incubators, makes its synchronization for 24 hours with serum free medium starved cells, after cultureThe A-H group Chinese medical extract combined treatments of 1 concentration of human umbilical cord mesenchymal stem cells association list.After handling 48h above, collect eachGroup cell, while cell culture supernatant is collected in -20 DEG C of preservations.
ELISA detects supernatant FN, type Ⅳ collagen content：Each group supernatant is extracted, is divided in micro reaction plate corresponding apertureSample to be tested is not added, then adds biotinylated antibody working solution 50txl.Mixing is vibrated, sets in 20-25 DEG C of environment and incubates120min.Liquid in hole is blotted, is fully washed 5 times with cleaning solution, it is dry.In addition to blank well, enzyme conjugates working solution is addedl00ul.Mixing is vibrated, sets in 20-25 DEG C of environment and incubates 30min.Liquid in hole is blotted, cleaning solution fully washs 5 times, doesIt is dry.Color developing agent each 50ul of A, B is added per hole, gently vibrates mixing, colour developing l0min is protected from light in 37 DEG C of environment.Terminate liquid is addedAfter 50ul, mixing is gently vibrated.Each hole absorbance value is measured with microplate reader (450nm wavelength).
The results are shown in Figure 5, and huMSCs can have renal lesions improvement effect on a cellular level, and FN and COL4 is presentedSignificant decrease, and the addition of Traditional Chinese drug mixture can have facilitation to huMSCs to the improvement of renal lesions.Especially it is worthIt is noted that demonstrate Radix Notoginseng in afore-mentioned test has inhibiting effect for huMSCs cell Proliferations, and low concentration Radix Notoginseng is extractedThe addition of liquid (final concentration 10-30ul/ml) may can reach in Chinese medical extract combination inhibits fibronectin and IVThe effect that Collagen Type VI generates.
6 human umbilical cord mesenchymal stem cells of experimental example combine the experiment of plurality of Chinese extract for treating
1) animal feeding
Cleaning grade male Wistar rat 60,200~220g of weight, by the sugar engineering technical research of country of Shandong UniversityThe heart provides.
All rats are the same as room sub-cage rearing, natural lighting, ad lib water inlet, 18~25 DEG C of raising temperature, relative humidity40~50%, avoid high-decibel noise；Ensure well-ventilated, 1h/ times, 3 times/d；Frequently change bedding and padding, 2-3 times/week.Raising is fitted for one weekStart to test after answering experimental enviroment.Fasting 12h before modeling, can't help water.
2) prepared by diabetes model
After rat adaptability raises 7d, it is deprived of food but not water 12h, single intraperitoneal injection STZ (50mgkg-1) induces glycosuriaIsometric 0.1mol/L citrate buffers are injected intraperitoneally in disease, control animals.It is examined with Instrument for Measuring Blood Sugar after injection STZ 3dBlood glucose is surveyed, glucose in urine is detected with Tes-Tape.Blood glucose >=16.7mmolL-1, glucose in urine >=+++ it is included in diabetic animal.
3) animal is grouped and reverses is administered at random
Experimental animal is grouped at random, respectively control group, model group, A-H Chinese medical extract combinatorial associations treatment group, every groupEach 6.
Wherein, control group is normal rat；The experimental subjects of model group be experimental procedure 2) in diabetic model rats, andNot treat；The experimental subjects of MSC treatment groups is experimental procedure 2) in diabetic model rats, and it is dry to give human umbilical cord mesenchymalCell therapy；The experimental subjects of A-I Chinese medical extract combinatorial associations treatment group be experimental procedure 2) in diabetic model rats, andGive human umbilical cord mesenchymal stem cells and plurality of Chinese extract combination therapy；
Each group rat oral gavage is given to be administered respectively daily, dosage is Chinese medicine composition mixed liquor：10ml/kg、MSC：2×106A/only.Chinese medicine composition mixed liquor is configured according to 2 concentration of table.
2 Chinese medicine mixed extract concentration of table configures
4) dextrose tolerance test
In testing for the 6th weekend, after Rat Fast 6 hours, gavage gives 20% glucose 1ml100g-1Weight is (quite2g·kg-1Weight), the blood glucose of 30,60 and 120min contains after measuring (0min) on an empty stomach and clothes sugar respectively with JPS-3+ types blood glucose meterAmount, as a result as shown in table 3 below：
Blood glucose value (the mmolL of 3 each group animal different time points of table-1)(x±s)
| Group | 0min | 30min | 60min | 120min |
| Control | 5.82±0.63 | 7.82±0.73 | 7.23±0.93 | 6.23±0.65 |
| Model | 25.04±4.51 | 36.50±2.83 | 30.12±5.46 | 27.54±4.65 |
| MSC | 23.58±3.45 | 28.31±3.55 | 23.89±4.12 | 19.86±4.11 |
| A | 21.86±0.84 | 24.84±2.76 | 21.58±0.32 | 19.01±2.94 |
| B | 22.98±0.16 | 25.68±3.15 | 20.40±2.02 | 17.96±3.53 |
| C | 19.16±2.71 | 24.15±0.54 | 22.29±3.93 | 20.00±2.69 |
| D | 20.50±0.08 | 24.81±2.46 | 14.58±0.92 | 12.37±3.42 |
| E | 21.31±3.6 | 22.14±3.37 | 16.90±3.69 | 14.20±3.66 |
| F | 20.87±1.05 | 24.44±2.55 | 14.01±0.41 | 13.17±1.23 |
| G | 22.06±0.35 | 23.48±3.46 | 16.50±1.76 | 14.88±0.34 |
| H | 20.04±3.29 | 23.44±2.93 | 17.35±1.63 | 14.42±1.96 |
| I | 20.12±3.18 | 24.76±1.80 | 18.26±0.20 | 15.38±1.54 |
By result in table 3 it is found that blank control group rat has normal glucose regulating power.Diabetes rats are in giving Portugal30~60min blood glucose is constantly in high level after grape sugar, is slightly decreased to blood glucose level when 120min, shows diabetes ratSugar tolerance is substantially reduced, and sugared regulative mechanism is impaired.It is notable using huMSCs reduction hypoglycemic effects in administration group, illustrate to break upHuMSCs have and potential improve diabetic nephropathy effect.And then more preferably, data are also bright for therapeutic effect by A-H groups cooperation huMSCsThe remarkable result that the aobvious Chinese medical extract combination cooperation huMSCs for embodying A-H reduces blood glucose level.Wherein embodied in A-C groupsRadix Astragali, Radix Salviae Miltiorrhizae and ginseng extract are respectively cooperating with Schisandra chinensis, Rhizoma Atractylodis Macrocephalae extract, close with cooperation huMSCs therapeutic effects；AndD-I groups are then shown, although Notogineng Extract inhibits huMSCs cellular levels proliferation, Radix Notoginseng and Radix Astragali, Radix Salviae Miltiorrhizae, ginseng threeTogether cooperation Schisandra chinensis and Rhizoma Atractylodis Macrocephalae Chinese medical extract combination can be obviously promoted improvement results of the huMSCs to renal lesions.
In conjunction with the above testing result it is found that in the present invention, can it be improved using A-H groups Chinese medical extract and huMSCsImpaired rat regulating and controlling blood sugar ability；Compared with Chinese medical extract and huMSCs are used alone, Chinese medical extract and huMSCsDrug combination is hypoglycemic, it is more excellent to improve renal function effect；Especially it is found that low concentration Notogineng Extract in Chinese medical extract groupTool, which improves significantly, in conjunction assists huMSCs to slow down injury of kidney, improves and reconciles blood glucose ability.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from the present invention'sMany other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claimsIncluding belonging to all such changes and modifications in the scope of the invention.