CN101647855A - Traditional Chinese medicine preparation for treating diabetic neuropathy and preparation method thereof - Google Patents

Abstract

The invention discloses a traditional Chinese medicine preparation for treating diabetic neuropathy, which is prepared from the traditional Chinese medicine raw materials of radix salviae miltiorrhizae, radix notoginseng and silynarin by ethanol leaching, water leaching and pulverizing. The traditional Chinese medicine preparation is a novel traditional Chinese medicine for treating diabetic nervecomplications, has an obvious protective action to diabetic hypoischium nerves, enhances the conduction velocity of diabetic sciatic nerves, has a certain lowering function to the enhanced level of MDA and sorbitol, has an obvious enhancement function to the activity of glutathione peroxidase GSH-PX, also has a certain enhancement function to the lowered activity of superoxide dismutase (SOD), lowers the level of saccharification blood red protein and the specific viscosity of the whole blood obviously, also has an obvious pain-relieving function, has a very good curative effect to the diabetic nerve complications, provides a novel application choice for extensive patients and has less variety of the raw materials and easy control of the product quality.

Description

A kind of Chinese medicine preparation for the treatment of diabetic neuropathy and preparation method thereof

Technical field

The present invention relates to technical field of Chinese medicines, particularly relate to a kind of Chinese medicine preparation for the treatment of diabetic neuropathy and preparation method thereof.

Background technology

Diabetes are one of modal chronic diseases, along with the raising of people's living standard, and the increase of aged tendency of population and fat incidence rate, the sickness rate of diabetes is ascendant trend year by year.Diabetes reach 2% at the sickness rate of China, and according to statistics, the diabetics made a definite diagnosis of China reaches 4,000 ten thousand, and with annual 1000000 speed increase.Diabetes are the dysbolismus diseases that cause disturbance of carbohydrate metabolism owing to hypoinsulinism, main feature is that blood glucose is too high, glycosuria, polyuria, polydipsia, polyphagia, become thin, tired, its main harm is its complication.

The complication of diabetes is divided into acute complications, chronic complicating diseases and neural complication, and neural complication is:

(1) sensory nerve: pain, numbness, hyperesthesia;

(2) nervus motorius: the dyskinesia that visible single neural paralysis causes, but local muscle atrophy;

(3) vegetative nerve: perspire unusual, blood pressure and changes in heart rate, urinary incontinence or urine retention, diarrhoea or constipation and sexual impotence etc.

Western medicine to treatment of diabetes based on blood sugar lowering, little to the chronic complicating diseases of diabetes therapeutic effect.And owing to contain a lot of chemical analysis strong to the human body toxic and side effects in the Western medicine, so it is serious to liver, the injury of kidney of human body to take Western medicine for a long time.

With take the blood sugar lowering Western medicine and compare, diabetics uses insulin can alleviate the organs and tissues such as liver, kidney of damage human body, still, is not that each diabetes patient can both use insulin, the diabetes patient who has can not use the diabetics of insulin, particularly hyperinsulinemia not use.And, if the insulin improper use is easy to generate hypoglycemia, therefore must could correctly use insulin according to the mensuration of islet function, also some diabetics is with insulin antibody, at this moment the insulinize effect is just not obvious, so note the diabetics that insulin antibody is arranged is eliminated the treatment of insulin resistant, use insulin just can reach the purpose of steady blood sugar lowering like this.What is more important is used insulin and to take Western medicine the same, has only blood sugar reducing function, does not have therapeutic efficiency, and diabetic complication is not had the obvious treatment effect.

At present, a lot of products that add the blood sugar lowering Western medicine in Chinese medicine are arranged on the market, such product is just bigger to patient's harm, blood glucose descended controlled rapidly after a lot of patients felt to take some tcm product, just think that this kind Chinese medicine is the good medicine of treatment diabetes, hardly realize Western medicine (blood sugar lowering) composition wherein must be arranged, because Western medicine (blood sugar lowering) wherein becomes doses not quite clear, when taking, the patient often taken dosage at double, so not only the harm to the internal organs organ is bigger, and cause hypoglycemia easily, take the generation of damage human organ and quickening diabetic complication for a long time, development, what is more important has delayed the best opportunity of patient treatment diabetes, has incured loss through delay the state of an illness, gives patient physiological and causes bigger misery psychologically.

Tcm product has the internal organs of adjusting, slow blood sugar reducing function, mainly be to distinguish that disease executing control, medication to the ill just, add and subtract with disease, play the effect for the treatment of both the principal and secondary aspects of a disease, adopt treatment by Chinese herbs glycosuria condition of disease to reach the effect of nursing one's health each internal organs organ dysfunction and raising patient autoimmunity from inside of human body, thereby fundamentally treated diabetes, the patient is got well.So the Chinese patent medicine of research and development treatment diabetes has important meaning to treatment of diabetes.

Summary of the invention

Technical problem to be solved by this invention provides a kind of Chinese medicine preparation for the treatment of diabetic neuropathy and preparation method thereof; this Chinese medicine preparation has no side effect, curative effect is good; fundamentally nurse one's health each internal organs organ dysfunction of human body; improve body immunity; the neuroprotective cell reaches the purpose for the treatment of both the principal and secondary aspects of a disease.

In order to solve the problems of the technologies described above, the present invention adopts following technical scheme:

According to listed as parts by weight, the Chinese medicine preparation that the present invention treats diabetic neuropathy is to be prepared from for 10～30 parts by 570～660 parts of raw material of Chinese medicine Radix Salviae Miltiorrhizaes, 100～170 parts of Radix Notoginseng and silymarin.

Preferably, the Chinese medicine preparation of above-mentioned treatment diabetic neuropathy is to be prepared from for 15～25 parts by 610～620 parts of raw material of Chinese medicine Radix Salviae Miltiorrhizaes, 125～145 parts of Radix Notoginseng and silymarin.

Best, the Chinese medicine preparation of aforementioned therapies diabetic neuropathy is to be prepared from for 20 parts by 612 parts of raw material of Chinese medicine Radix Salviae Miltiorrhizaes, 136 parts of Radix Notoginseng and silymarin.

The present invention also provides a kind of preferred manufacturing procedure of Chinese medicine preparation of aforementioned therapies diabetic neuropathy: gets Radix Notoginseng and pulverizes, and standby, get Radix Salviae Miltiorrhizae, add ethanol extraction, filter filtrate recycling ethanol, the medicinal residues extracting in water, filter, concentrate, merge concentrated solution and ethanol extract, dry, pulverize, add Radix Notoginseng fine powder and silymarin, preparation process is made various dosage forms routinely.

Concrete, above-mentioned preparation method is: gets Radix Notoginseng powder and is broken into fine powder, and standby, get Radix Salviae Miltiorrhizae, add 4～8 times of amount 75%～95% ethanol extractions 1～3 time, each 1～3 hour, filter, merging filtrate, filtrate recycling ethanol, medicinal residues add 6～12 times of water gagings and decoct extraction 2～4 times, each 0.5～2 hour, filter merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merges concentrated solution and ethanol extract, drying, pulverize, add Radix Notoginseng fine powder and silymarin, preparation process is made various dosage forms routinely.

The preparation method of capsule of the present invention is: gets Radix Notoginseng powder and is broken into fine powder, and standby, get Radix Salviae Miltiorrhizae, add 6 times of amount 85% ethanol extractions 2 times, each 2 hours, filter merging filtrate, filtrate recycling ethanol, medicinal residues add 8 times of water gagings and decoct extraction 3 times, each 1 hour, filter, merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merges concentrated solution and ethanol extract ,-0.06MPa～-the 0.08MPa condition under microwave vacuum drying become dry extract, pulverize, add Radix Notoginseng fine powder and silymarin, add adjuvant, mixing, incapsulate, promptly get capsule.

The function of Chinese medicine preparation of the present invention cures mainly: have supplementing QI and nourishing YIN, set upright joint venture, row stagnates and promotes blood circulation, the merit of activating collaterals to relieve pain, be used for diabetic neuropathy and comprise diseases such as peripheral nervous inflammation, diabetes autonomic nervous dysfunction.

In we: Radix Salviae Miltiorrhizae, the dry root and rhizome of labiate Radix Salviae Miltiorrhizae (Salvia miltiorrhiza Bge.), hardship, be slightly cold, GUIXIN, Liver Channel have stasis-dispelling and pain-killing, promoting blood circulation to restore menstrual flow, the effect of the relieving restlessness that clears away heart-fire is used for menoxenia, the amenorrhea dysmenorrhea, lump in the abdomen, breast ventral spine pain, pyretic arthralgia pain, skin infection swells and ache, dysphoria and insomnia, hepatosplenomegaly, diseases such as angina pectoris.Radix Notoginseng (Panax Notoginseng) is used as medicine with root, root stock, is rare Chinese medicine, and sweet and slightly bitter taste is warm in nature, returns liver, stomach warp, but gives birth to hemostasia and dissipation blood stasis, reducing swelling and alleviating pain.Silymarin (Silymarin), faint yellow or pale brown toner end, no special odor, be through the refining flavone compound that forms of scientific method by Compositae medicinal plants Herba Silybi mariani seed, its main component is silibinin, Isosilybin, Silychristin, it is peaceful etc. to fly the water Ji, have protect the liver, radioprotective and effect for reducing blood fat.More than three flavor Chinese medicine Synergistics, play supplementing QI and nourishing YIN altogether, set upright joint venture, row is stagnantly promoted blood circulation, the effect of activating collaterals to relieve pain.

Determining of test example one, extraction process and molding technological condition

1, Radix Salviae Miltiorrhizae extraction process

1.1 Radix Salviae Miltiorrhizae alcohol extraction process

Alcohol extraction process adopts Radix Salviae Miltiorrhizae alcohol extraction orthogonal optimum seeking technology.Taking by weighing red rooted salvia 60g, is that the examination factor is all extracted 2 times with concentration of alcohol, return time, amount of alcohol, carries out L ₉(3 ⁴) orthogonal test, and be evaluation index with tanshinone content, filter out optimised process.

1.1.1 the tanshinone assay is measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D).

Chromatographic condition and system suitability test are filler with the carbon octadecylsilane chemically bonded silica; Methanol-water (75: 25) is a mobile phase; The detection wavelength is 270nm, and theoretical cam curve is pressed Tanshinone I I _AThe peak meter should be not less than 3000.

The preparation precision of reference substance solution takes by weighing Tanshinone I I _AReference substance is an amount of, puts in the 10ml brown bottle, adds methanol to scale, shakes up, and makes the solution that every 1ml contains 0.18mg, promptly.

The preparation of need testing solution is got alcohol extract and is directly filtered, and gets the filtrate of continuing, promptly.

Accurate reference substance solution, each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

1.1.2 test arrangement and the results are shown in Table 1, table 2, table 3:

Table 1. empirical factor level

Table 2. is with Tanshinone I I _AOrthogonal experiment data for the preferred extraction process by water of index

The data intuitive analysis:

Compare the meansigma methods of each factor varying level experimental result, to factor A, maximum horizontal is 2, and to factor B, maximum horizontal is 2, and to factor C, maximum horizontal is 2, and factor D is a blank column.The size of each factor extreme difference R value relatively draws and to the descending order of index tanshinone content influence factor is: C＞B＞A.

Variance analysis

Table 3. is with Tanshinone I I _AOrthogonal experiment data variance analysis for the preferred alcohol extraction process of index

F0.05(2，2)＝19，F0.01(2，2)＝90

The result shows: three factors have significant difference.Determine alcohol extraction process according to intuitive analysis and variance analysis: 6 times of amounts of 85% ethanol, reflux, extract, 2 times, each 2 hours.Result of the test shows Tanshinone I I _AThe rate of transform is 64.01%, and the total solid substance yield is 15.79%, RSD=3.3%.

1.2 Radix Salviae Miltiorrhizae extraction process by water

Last medicinal residues carried out orthogonal optimum seeking after extraction process by water adopted the Radix Salviae Miltiorrhizae alcohol extraction.Equal with amount of water, extraction time, extraction time examination factor, carry out L ₉(3 ⁴) orthogonal test, and be evaluation index with the content of danshinolic acid B, filter out optimised process.

1.2.1 content of danshinolic acid B is measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D) and is measured.

Chromatographic condition and system suitability test are filler with the carbon octadecylsilane chemically bonded silica; Methanol-acetonitrile-formic acid-water (30: 10: 1: 59) be mobile phase; The detection wavelength is 286nm, and theoretical cam curve should be not less than 3000 by the salvianolic acid B peak.

It is an amount of that the preparation precision of reference substance solution takes by weighing the salvianolic acid B reference substance, adds 75% dissolve with methanol, makes the solution that every 1ml contains 0.15mg, promptly.

The preparation water intaking extracting solution of need testing solution directly filters, and gets the filtrate of continuing, promptly.

Accurate reference substance solution, each the 5 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

1.2.2 test arrangement and the results are shown in Table 4, table 5, table 6:

Table 4 empirical factor level

Table 5 is the orthogonal experiment data of the preferred extraction process by water of index with the salvianolic acid B

The data intuitive analysis

Compare the meansigma methods of each factor varying level experimental result, to factor A, maximum horizontal is 3, and to factor B, maximum horizontal is 1, and to factor C, maximum horizontal is 1, and factor D is a blank column.The size of each factor extreme difference R value relatively draws and to the descending order of index content of danshinolic acid B influence factor is: C＞B＞A.

Variance analysis

Table 6. is the orthogonal experiment data variance analysis of the preferred extraction process by water of index with the salvianolic acid B

Factor	??SS	Degree of freedom	The F value
				??A	??0.03	??2	??0.23
??B	??0.06	??2	??0.52
				??C	??0.48	??2	??3.94
Error	??0.12	??2
				Amount to	??0.69	??8

F0.05 (2,2)=19, there was no significant difference.

The result shows: three factor there was no significant differences.Determine extraction process by water according to intuitive analysis: 8 times of water gagings decoct each 1 hour 3 times.Result of the test shows that the salvianolic acid B rate of transform is 88.0%, and the total solid substance yield is 15.79%, RSD=0.3%.

1.3 total solid matters yield

Total solid matters yield (receiving the cream rate) is measured the accurate extracting solution 25ml that draws, place the evaporating dish that is dried to constant weight, volatilize, in 105 ℃ of dryings 5 hours, move in the exsiccator, cooled off 30 minutes, in 105 ℃ dry again 1 hour, move in the exsiccator, cooling, weight decided in accurate rapidly title, calculates, and promptly gets the total solid substance yield.

The table 7 alcohol extraction process tanshinone rate of transform, total solid yield

The table 8 extraction process by water salvianolic acid B rate of transform and total solid yield

2, concentrate with drying process preferably

Need to carry out the concentrate drying rear plastic because former side is prepared into Chinese medicine preparation, therefore, comparative study carried out in the variation of tanshinone, salvianolic acid B before and after the concentrate drying.

2.1 the condition of concentrating is to the influence of capsule

Get the 6120g red rooted salvia, extract by the technology of working out, filtrate adopts the rotating thin film evaporimeter to concentrate, alcohol extract is at vacuum-0.07MPa, 55 ℃ of bath temperatures be concentrated into ethanol wave to the greatest extent solution 1500ml, the water extract is at vacuum-0.07MPa, and bath temperature is concentrated into relative density 1.05 for 65 ℃, concentrate 1600ml solution.Two parts of solution are mixed.The content of sampling and measuring tanshinone, salvianolic acid B calculates the rate of transform.

Result of the test shows that the tanshinone rate of transform is 62.42% before and after concentrating, and average loss rate is 1.59%, and the salvianolic acid B rate of transform is 78.41% before and after concentrating, and average loss rate is 10.35%, illustrates that concentration technology does not have obvious influence to the capsule quality.

2.2 different drying conditions relatively

Merge above concentrated solution, be divided into 6 parts, every part of 500ml, dry under the different condition, sampling and measuring tanshinone, content of danshinolic acid B the results are shown in Table 9.

The different dry condition effect of table 9

Result of the test shows that the constant pressure and dry time is long, and tanshinone, the salvianolic acid B rate of transform are lower than microwave vacuum drying.The vacuum drying time is short, tanshinone, salvianolic acid B rate of transform height, and the sample quality is loose, is easy to pulverize.Under the microwave vacuum condition ,-0.08MPa and-the 0.06MPa drying has certain influence to tanshinone, the salvianolic acid B rate of transform, so can-0.06MPa～-the 0.08MPa condition under vacuum drying.

3, moulding process

Medicated powder is pulverized after drying, crosses sieve No. 5.Radix Notoginseng powder is broken into fine powder, together adds in the above-mentioned fine powder with silymarin, and mixing incapsulates, promptly.

Test example two, pharmacodynamic experiment

Summary: zoopery is the result show: easypro conduction velocity and the action potential refractory stage to diabetes os hypogastroidale nerve of (1) meridians has significant protective effect; can obviously improve the sciatic conduction velocity of diabetes; shorten its action potential refractory stage, and sciatic pathological change is had the improvement effect.(2) malonaldehyde (MDA) and the sorbitol level that diabetic sciatic nerve is raise has certain reduction effect, to the activity effect of being significantly improved of glutathion peroxidase GSH PX, to superoxide dismutase (SOD) activity that the descends effect that also improves.(3) meridians relax and can obviously reduce the blood red egg level of saccharifying and the whole blood contrast viscosity of diabetes rat.(4) meridians relax and have significant analgesia role

Test objective: the function of " meridians relieving capsule " (capsule of the present invention) cures mainly and is supplementing QI and nourishing YIN, sets upright joint venture, and row stagnates and promotes blood circulation the merit of activating collaterals to relieve pain.Be used for diabetic neuropathy and comprise peripheral nervous inflammation, diabetes autonomic nervous dysfunction.Cure mainly according to its function; its protective effect to diabetic peripheral neuropathy is observed in this experiment on the streptozotocin diabetes animal model; observe its improvement effect to the neural polyhydric alcohol metabolic pathway that occurs of diabetes os hypogastroidale and SOD, MDA, the disorder of GSH-PX level; observe its influence, and observe its antiinflammatory, analgesic activity diabetic animal hemorheology and glycolated hemoglobin.For its clinical practice provides experimental basis.

Material and key instrument

1, is subjected to the reagent and the unit of providing

(1) meridians relieving capsule, being tried thing is its content medicated powder.Every capsules is adorned 0.35 gram medicated powder, and every capsules contains primary crude drug 1.0 gram (i.e. 0.35 gram medicated powder be equivalent to primary crude drug 1.0 restrain), intends consumption clinical every day and is equivalent to primary crude drug 9～12g/60kg and (promptly is equivalent to medicated powder 3.15～4.2g/60kg).Guizhou Province Chinese medicine preparation research and development centre provides lot number: 20050601.

(2) TANGMAIKANG KELI, function cures mainly: replenishing YIN and removing heat, blood circulation promoting and blood stasis dispelling, QI invigorating controlling nocturnal emission with astringent drugs.Be used for the diabetes deficiency of both QI and YIN Blood stasis of holding concurrently, disease is seen fatigue and weakness, deficiency of QI with disinclination to talk, spontaneous perspiration, night sweat, dysphoria with feverish sensation in the chest palms and soles, thirst and liking drink, vexed pain, numb limbs and tense tendons or twinge, constipation etc. and diabetes type II and complication thereof the person that sees the above-mentioned symptom in the heart.Usage and dosage: oral, one time 1 bag, 3 times on the one.Dosage form specification: granule, every bag 5 gram, 10 bags in every box.Zhonghui Pharmacy Co., Ltd., Chengdu, lot number: 050617.

2, other medicine and reagent

(1) morphine hydrochloride injection, 1ml: 10mg/10 props up, Shenyang No. 1 Pharmaceutical Factory production, lot number: 040201 (valid until 2009.01).

(2) protamine zinc insulin injection, 10ml: 400u/2 props up, and Chinese Jiangsu ten thousand helps biochemical medical limited company to produce lot number: 0306208 (valid until 2005.06).

(3) maleate chlorpheniramine injection (chlorphenamine), 1ml: 10mg/10 props up, Wuxi City the 7th pharmaceutical Co. Ltd, lot number: 03031213 (valid until 2006.03).

(4) dexamethasone sodium phosphate injection, 1ml: 5mg/10 props up, Wuxi City the 7th pharmaceutical Co. Ltd, lot number: 04082013 (valid until 2006.07).

(5) streptozotocin (Streptozotocin, STZ); U.S. Sigma product, 98%HPLC, S0130-1G, 015K1279.

(6) malonaldehyde test kit (MDA), superoxide dismutase test kit (SOD), glutathion peroxidase test kit (GSH-PX), glycolated hemoglobin test kit (GHb), be Nanjing and build up bio-engineering research and produce, lot number is 20051107.

(7) Oleum Tiglii is by the preparation of pharmacology teaching and research room, lot number: 20040511.

(8) sorbitol, 25 gram/bottles, Chemical Reagent Co., Ltd., Sinopharm Group, lot number: T20050815.

(9) periodic acid, 100 gram/bottles, Chemical Reagent Co., Ltd., Sinopharm Group, lot number: F20050829.

(10) chromotropic acid, 25 gram/bottles, Tianjin recovery fine chemistry industry institute, lot number: 20041222.

(11) heparin sodium injection, 2ml: 12500u/10 props up, Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd, lot number: 030803 (valid until 2006.07).

(12) arsenic oxide, the 250g/ bottle, Chemical Reagent Co., Ltd., Sinopharm Group, being put on record to preserve by my school chemistry teaching and research room provides.

3, animal

(1) male Wistar rat, body weight 200～250g;

(2) Kunming mouse, body weight 18～25g, male and female dual-purpose.

4, key instrument

One-Touch blood sugar detection instrument, BL-420 biological function experimental system, 721 spectrophotometers, TDL-5 centrifuge, LG-R-80A blood viscosity instrument.

Method is selected:

Cure mainly according to the function that is subjected to reagent, carry out the pharmacodynamic study of following project:

1, to the influence of diabetes rat peripheral neuropathy

(1) to the influence of rat sciatic nerve conduction velocity and action potential refractory stage

(2) sciatic nerve is made histological observation

(3) observation of sciatic nerve sorbitol content, mda content (MDA), superoxide dismutase activity (SOD), activity of glutathione peroxidase (GSH-PX)

2, to the influence of diabetic mice peripheral neuropathy

3, to the influence of blood glucose in diabetic rats, glycolated hemoglobin

4, to the influence of rat diabetes hemorheology index

5, analgesic activity

Experimental control:

1, normal control group: give normal saline;

2, model control group: give normal saline;

3, positive control drug: be Chinese medicine matched group (TANGMAIKANG KELI) and western medicine group (matched groups such as insulin, morphine, dexamethasone, chlorphenamine).

Route of administration, administration volume and dosage:

1, route of administration:

Meridians relax and TANGMAIKANG KELI is the administration of filling stomach (i.g).

2, gastric infusion volume:

Rat 10ml/kg, mice 20ml/kg

3, dosage:

Meridians relax: take by weighing an amount of medicated powder, prepare to desired concn (guaranteeing the administration constancy of volume) with an amount of dissolved in distilled water.

TANGMAIKANG KELI: directly get its granule, prepare to desired concn (guaranteeing the administration constancy of volume) with dissolved in distilled water.

4, other contrast medicines:

Insulin: every 50 μ l of rat direct injection; Mice 10u/kg (getting 0.25ml with 10 times of injecting normal saline dilutions, injection 2.5ml/kg);

Morphine: stock solution is diluted 10 times (containing morphine 1mg/ml) with the injection normal saline, gives mouse peritoneal injection 10ml/kg (10mg/kg).

Dexamethasone: stock solution is diluted 1 times (containing dexamethasone 2.5mg/ml) with the injection normal saline, gives rat skin lower injection 1.6ml/kg (4mg/kg).

Chlorphenamine: stock solution is diluted 10 times (containing chlorphenamine 1mg/ml) with the injection normal saline, gives rats by intraperitoneal injection 4ml/kg (4mg/kg).

Date processing:

Adopt " Chinese medicine encyclopedia medicostatistics " statistical package (second edition) of vital statistics teaching and research room of Huaxi Medical Univ to carry out relevant check.Continuous data is represented with x ± s.

Experimental technique and result

1, to the influence of diabetic sciatic nerve conduction velocity (be called for short MNCV, down with), action potential refractory stage and pathological change thereof

Extracting male Wistar rat, body weight 200～250g, fasting 12h before the experiment, lumbar injection streptozotocin (STZ then, 48mg/kg, face with preceding and prepare with citrate buffer) modeling, with the fasting glucose of One-Touch blood sugar detection instrument mensuration animal fasting 12h, blood sugar level＞15mmol/L person is a diabetes rat behind the 72h.

Get diabetes rat, be divided into model control group, Western medicine positive controls (insulin, intramuscular injection), Chinese medicine positive controls (TANGMAIKANG), the easypro high dose group of meridians, the easypro middle dosage group of meridians, the easypro low dose group of meridians at random.Other gets rat and does not inject streptozotocin, as the normal control group.After the diabetes 30 days, respectively get 5 rats from model control group, normal control group respectively and measure MNCV, change trend occurs, can begin administration, every day 1 time, continuous 60 days as model group MNCV and action potential refractory stage.And after administration, measured every group body weight on the 15th, 30,45,60 day, the amount of drinking water of record every day during the administration.Administration is respectively organized after 30 days and is got part rat mensuration MNCV and action potential refractory stage, and assay method and step are as follows:

Behind the rat anesthesia, the ventricumbent position is fixed, fully expose right sciatic nerves, stimulating electrode places the sciatic nerve at incisura ischiadica place to spread out of position (near-end), recording electrode places tibia internal malleolus place (far-end), connects BL-420 biological function experimental system, selects suitable stimulus parameter (pulse, the wide 0.1ms of ripple, 1.5 times of threshold values of stimulus intensity, more than the stimulus intervals 5s), control room temperature (20 ± 0.5 ℃) keeps 37 ℃ of rat temperatures.From computer recording sciatic nerve action potential refractory stage (incubation period), every rat repetitive stimulation 3 times, get its meansigma methods, accurately measure stimulating electrode to the distance between the recording electrode, the substitution formula: sciatic nerve conduction velocity (m/s)=stimulating electrode is to the distance between the recording electrode/action potential refractory stage.

After the administration 60 days, each group residue rat is observed its MNCV and action potential refractory stage, and sciatic nerve is carried out histological observation.The result is as follows:

(1) to the influence of rat body weight, amount of drinking water

The result shows, meridians each dosage group of relaxing there is no protective effect to the weight loss and the amount of drinking water increase of diabetes rat, sees Table 1, table 2 (a) and table 2 (b).

The easypro influence to the diabetes rat body weight of table 1 meridians (x ± s, g, n=5)

Compare with the normal control group: * * P＜0.01

The easypro influence to diabetes rat amount of drinking water every day of table 2 (a) meridians (x ± s, ml/, n=5)

Compare with the normal control group: * * P＜0.01

The easypro influence to diabetes rat amount of drinking water every day of table 2 (b) meridians (x ± s, ml/, n=5)

Group	The 31st～40 day	The 41st～50 day	The 51st～60 day
				The normal control group	??40.40±0.70(10)	??40.65±0.71(10)	??41.55±0.60(10)
Model control group	??75.60±3.64(15) **	??78.53±7.57(14) **	??82.17±4.31(12) ?? **
				The insulin matched group	??74.62±7.34(13) **	??76.50±8.69(12) **	??83.40±4.72(10) ?? **
The TANGMAIKANG matched group	??73.87±4.86(15) **	??75.54±6.59(13) **	??81.00±5.04(11) ?? **
				The meridians high dose that relaxes	??75.00±7.58(14) **	??77.83±7.37(12) **	??83.30±4.88(10) ?? **
Dosage during meridians relax	??76.29±5.35(14) **	??78.00±4.96(13) **	??81.25±4.80(12) ?? **
				The meridians low dosage that relaxes	??76.86±7.30(14) **	??79.62±6.61(13) **	??81.09±4.32(11) ?? **

Compare with the normal control group: * * P＜0.01

(2) to the influence of MNCV and action potential refractory stage

(1) result shows, diabetes are after 30 days, the rat MNCV of normal control group be 30.29 ± 5.93 (x ± s, n=5), the action potential refractory stage be 0.51 ± 0.10 (x ± s, n=5); Diabetic model group rat MNCV be 22.60 ± 5.55 (x ± s, n=5), the action potential refractory stage be 0.70 ± 0.19 (x ± s, n=5).Compare with the normal control group, the action potential prolonged refractory period 37.25% of diabetes rat, MNCV occur descending 25.39%, see Table 3.

(2) drug treatment is after 30 days, easypro each the dosage group of meridians all has some improvement to the MNCV decline of diabetes rat, relatively improves 11.54% with model group, and the action potential refractory stage is shown certain shortening effect, relatively shorten 11.71% with model group, see Table 3.Behind the drug treatment 60 days, the easypro high dose group of meridians can obviously shorten the sciatic nerve action potential refractory stage time, and MNCV is slowed down to improve significantly, and sees Table 4.

The table 3 meridians administration influence to diabetic sciatic nerve conduction velocity and action potential refractory stage in the 30 days (x ± s) that relaxes

Group	Dosage	Number of animals	Action potential refractory stage (ms)	Conduction velocity (m/s)
					The normal control group		??10	??0.51±0.09	??31.26±5.68
Model control group		??10	??0.67±0.14 **	??23.37±4.58 **
					The insulin matched group	2u/ only	??10	??0.53±0.14 △△	??30.58±9.07 △
The TANGMAIKANG matched group	??2.5g/kg	??10	??0.56±0.13 △	??28.33±7.96
					The meridians high dose group of relaxing	2g crude drug/kg	??10	??0.59±0.09 △	??26.06±4.13 *
Dosage group during meridians relax	1g crude drug/kg	??10	??0.62±0.10 *	??24.63±4.00 **
					The meridians low dose group of relaxing	0.5g crude drug/kg	??10	??0.63±0.14 *	??25.08±6.36 *

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05; △ △ P＜0.01

The table 4 meridians administration influence to diabetic sciatic nerve conduction velocity and action potential refractory stage in the 60 days (x ± s) that relaxes

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05; △ △ P＜0.01

(3) influence that the sciatic nerve pathology are changed

After the administration after 60 days, each group diabetic sciatic nerve has been carried out histological observation, and histology's pathological change rank marked: (1) sciatic adventitia, bundle film, epilemma structure are clear, the Schwann cell form is normal, a matter blood capillary and nerve fiber aixs cylinder are all normal, and keeping the score is 0 minute; (2) matter internal zone dividing territory blood capillary intimal thickening between, keeping the score is 1 minute; (3) matter internal zone dividing territory blood capillary intimal thickening between, its peripheral nerve sheath disappearance, performance demyelination phenomenon, keeping the score is 2 minutes; (4) matter internal zone dividing territory blood capillary intimal thickening between, its peripheral nerve sheath disappearance, demyelination, corresponding aixs cylinder mild swelling etc., keeping the score is 3 minutes.

The result shows, the sciatic adventitia of rats in normal control group, bundle film, epilemma structure are clear, and the Schwann cell form is normal, a matter blood capillary and nerve fiber aixs cylinder are all normal; A matter internal zone dividing territory blood capillary intimal thickening all appears in the diabetic model group rat sciatic nerve, its peripheral nerve sheath disappearance, performance demyelination phenomenon, changes such as corresponding aixs cylinder mild swelling; Histology's pathological change rank scoring of insulin, TANGMAIKANG, easypro each the administration group of meridians decreases, and the easypro pathological change to diabetic sciatic nerve of prompting meridians has the certain protection effect, sees Table 5.

The administration protective effect to the diabetic sciatic nerve pathological change in 60 days of relaxing of table 5 meridians

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05; △ △ P＜0.01

2, to the influence of diabetic mice sciatic nerve conduction velocity, action potential refractory stage

Male mice intravenous injection 200mg/kg STZ measured tail vein sugar with ONE TOUCH blood sugar test paper after 72 hours, was diabetic mice greater than the mice of 15mol/L (inconsistent with the back).Diabetic mice is divided into the high, medium and low dosage group of diabetes model matched group, insulin matched group, Chinese medicine positive controls and meridians relieving capsule at random.About every group 20.Other get do not inject STZ 10 mices as the normal control group.Promptly begin administration after the grouping, every day 1 time, continuous 60 days.Put to death mouse assay mice sciatic nerve conduction velocity and action potential refractory stage in the 61st day, assay method and step are as follows:

Behind the mouse anesthesia, the ventricumbent position is fixed, fully expose right sciatic nerves, stimulating electrode places the sciatic nerve at incisura ischiadica place to spread out of position (near-end), recording electrode places tibia internal malleolus place (far-end), connects BL-420 biological function experimental system, selects suitable stimulus parameter (pulse, the wide 0.1ms of ripple, 1.5 times of threshold values of stimulus intensity, more than the stimulus intervals 5s), keep 25 ℃ of room temperatures, keep 37 ℃ of mouse temperatures.From computer recording sciatic nerve action potential refractory stage (is incubation period, ms), every mice repetitive stimulation 3 times, get its meansigma methods, accurately measure stimulating electrode to the distance between the recording electrode (cm), the substitution formula: sciatic nerve conduction velocity (m/s)=stimulating electrode is to the distance between the recording electrode/action potential refractory stage.

The result shows that the easypro high dose group of meridians can obviously shorten the sciatic action potential refractory stage time of diabetic mice, and MNCV is slowed down to improve significantly, and sees Table 6.

The table 6 meridians administration influence to diabetic mice sciatic nerve conduction velocity and action potential refractory stage in the 60 days (x ± s) that relaxes

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05

3, to the influence of diabetic sciatic nerve sorbitol level

Extracting male Wistar rat, body weight 200～250g, fasting 12h before the experiment, lumbar injection streptozotocin (48mg/kg then, face with preceding and prepare with citrate buffer) modeling, measure the fasting glucose of animal fasting 12h behind the 72h with One-Touch blood sugar detection instrument, blood sugar level＞15mmol/L person is a diabetes rat.

Get diabetes rat, be divided into model control group, Western medicine positive controls (insulin, intramuscular injection), Chinese medicine positive controls (TANGMAIKANG), the easypro high dose group of meridians, the easypro middle dosage group of meridians, the easypro low dose group of meridians at random.Other gets rat and does not inject streptozotocin, as the normal control group.After the diabetes modeling 30 days, each group beginning administration, every day 1 time, continuous 60 days, after 60 days, get sciatic nerve homogenate in administration, get the chemical determination sorbitol level of supernatant by list of references.

The result shows that diabetic sciatic nerve sorbitol level obviously raises, and high dose, middle dosage and low dose group meridians relax the horizontal reduction rate of diabetic sciatic nerve sorbitol is respectively 29.74,24.38,17.47%.See Table 7.

The easypro influence to diabetic sciatic nerve sorbitol level of table 7 meridians (x ± s)

Group	Dosage	Number of animals	Sorbitol content (umol/l)	Reduction rate (%) (comparing) with model group
					The normal control group		??10	??2.25±1.76
Model control group		??12	??4.50±2.38 *
					The insulin matched group	2u/ only	??10	??2.42±1.95 △	?46.33
The TANGMAIKANG matched group	??2.5g/kg	??11	??3.22±1.24	?28.61
					The meridians high dose group of relaxing	2g crude drug/kg	??10	??3.17±1.54	?29.74
Dosage group during meridians relax	1g crude drug/kg	??12	??3.41±1.73	?24.38
					The meridians low dose group of relaxing	0.5g crude drug/kg	??11	??3.72±1.66 *	?17.47

Compare with the normal control group: * P＜0.05; Compare with model control group: △ P＜0.05

4, to the influence of diabetic sciatic nerve SOD, MDA, GSH-PX level

Extracting male Wistar rat, body weight 200～250g, fasting 12h before the experiment, lumbar injection streptozotocin (48mg/kg then, face with preceding and prepare with citrate buffer) modeling, measure the fasting glucose of animal fasting 12h behind the 72h with One-Touch blood sugar detection instrument, blood sugar level＞15mmol/L person is a diabetes rat.

Get diabetes rat, be divided into model control group, Western medicine positive controls (insulin, intramuscular injection), Chinese medicine positive controls (TANGMAIKANG), the easypro high dose group of meridians, the easypro middle dosage group of meridians, the easypro low dose group of meridians at random.Other gets rat and does not inject streptozotocin, as the normal control group.After the diabetes modeling 30 days, each group beginning administration, every day 1 time, continuous 60 days, in administration after 60 days, get sciatic nerve homogenate, get supernatant and measure sciatic nerve mda content (MDA), superoxide dismutase activity (SOD), glutathion peroxidase (GSH-PX) activity level by kit method and literature method.

The MDA test philosophy: the MDA in the lipid peroxide catabolite can with sulfo-barbital (TBA) condensation, form red product, at the 532nm place maximum absorption peak is arranged.The SOD test philosophy: xanthine and xanthine oxidase reaction can produce ultra-oxygen anion free radical (O ₂ ^-), latter's oxidation hydroxyl forms nitrite, is aubergine under the effect of developer, surveys its absorbance with visible spectrophotometer.When sample contains SOD, then ultra-oxygen anion free radical there is narrow spectrum inhibitory action, the nitrite of formation is reduced, absorbance reduces during colorimetric, can calculate SOD vigor in the sample by formula.

The GSH-PX test philosophy: reduced glutathion (GSH) and the effect of dithio dinitrobenzoic acid generate five sulfo-dinitrobenzoic acid aniones, present more stable yellow.Glutathion peroxidase in the body (GSH-PX) specifically catalysis GSH to hydrogen peroxide (H ₂O ₂) reduction reaction, form H ₂O and oxidized form of glutathione.Measure the consumption of reduced glutathion (GSH) in the enzymatic reaction, then can calculate the vigor of glutathion peroxidase.

The result shows that diabetic sciatic nerve mda content (MDA) level obviously raises, and activity of glutathione peroxidase (GSH-PX), superoxide dismutase activity (SOD) obviously reduce.

With diabetic model group relatively, relax high dose, middle dosage and low dose group of meridians has the reduction effect to the MDA level of diabetic sciatic nerve rising, reduction rate is respectively 21.32,15.23,10.78%; To the activity effect of being significantly improved of GSH-PX, the raising rate is respectively 18.82,12.81,9.90%; Relax high dose, middle dosage of meridians also has the raising effect to the SOD activity that descends, and the raising rate is respectively 14.00,11.79%; See Table 8.

The easypro influence to diabetic sciatic nerve MDA, GSH-PX, SOD of table 8 meridians (x ± s)

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05

5, to the influence of blood glucose in diabetic rats, saccharification hemoglobin content level

Extracting male Wistar rat, body weight 200～250g, fasting 12h before the experiment, lumbar injection streptozotocin (48mg/kg then, face with preceding and prepare with citrate buffer) modeling, measure the fasting glucose of animal fasting 12h behind the 72h with One-Touch blood sugar detection instrument, blood sugar level＞15mmol/L person is a diabetes rat.

Get diabetes rat, be divided into model control group, Western medicine positive controls (insulin, intramuscular injection), Chinese medicine positive controls (TANGMAIKANG), the easypro high dose group of meridians, the easypro middle dosage group of meridians, the easypro low dose group of meridians at random.Other gets rat and does not inject streptozotocin, as the normal control group.Promptly begin administration after the grouping, every day 1 time, continuous 60 days, measured rat fasting blood-glucose in the 61st day, with 1% anticoagulant heparin whole blood, 3～4ml, 1000 rev/mins centrifugal 10 minutes, abandon supernatant and stay the precipitation erythrocyte, wash as stated above 2～3 times with normal saline, (principle is that the glycolated hemoglobin that has the ketoamine key in the hemoglobin heats in sour environment, makes the hexose partial dehydration, generates the 5 hydroxymethyl furfural chemical compound to measure saccharification hemoglobin content by test kit description method and literature method again, the latter can be yellow with sulfo-barbital (TBA) reaction, carries out colorimetric assay)

The result shows that administration was compared with model group after 60 days, and the meridians of high dose and middle dosage group relax the trend of reduction to the diabetes rat fasting glucose, and reduction rate is respectively 20.4%, 13.3%.The meridians of high dose group relax can obviously reduce diabetes rat glycolated hemoglobin level, sees Table 9.

The easypro influence to blood glucose in diabetic rats, glycolated hemoglobin of table 9 meridians (x ± s)

Group	Dosage	Number of animals	Blood glucose (mmol/l)	Glycolated hemoglobin (g/ml)
					Normal control group group		??10	??4.96±1.10	??18.68±6.28
Model control group		??12	??19.14±5.37 **	??31.01±13.88 *
					The insulin matched group	2u/ only	??10	??11.95±4.92 **△△	??19.53±5.58 △
The TANGMAIKANG matched group	??2.5g/kg	??11	??15.42±4.87 **	??22.90±6.31
					The meridians high dose group of relaxing	2g crude drug/kg	??10	??15.24±6.02 **	??20.84±4.45 △
Dosage group during meridians relax	1g crude drug/kg	??12	??16.59±5.10 **	??24.24±5.92 *
					The meridians low dose group of relaxing	0.5g crude drug/kg	??11	??20.46±5.81 **	??26.89±10.37 *

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05

6, to the hemorheological influence of diabetes rat

Extracting male Wistar rat, body weight 200～250g, fasting 12h before the experiment, lumbar injection streptozotocin (48mg/kg then, face with preceding and prepare with citrate buffer) modeling, measure the fasting glucose of animal fasting 12h behind the 72h with One-Touch blood sugar detection instrument, blood sugar level＞15mmol/L person is a diabetes rat.

Get diabetes rat, be divided into model control group, Western medicine positive controls (insulin), Chinese medicine positive controls (TANGMAIKANG), the easypro high dose group of meridians, the easypro middle dosage group of meridians, the easypro low dose group of meridians at random.Other gets rat and does not inject streptozotocin, as the normal control group.Promptly begin administration after the grouping, every day 1 time, continuous 60 days, adopted whole blood 1% anticoagulant heparin in the 61st day, on the LG-R-80A blood viscosity instrument, measure hemorheology.

The result shows, relax high dose, middle dosage group of meridians can obviously reduce the whole blood contrast viscosity of the high shear rate of diabetes rat blood whole blood, middle shear rate, low shear rate, sees Table 10.

The easypro influence to diabetes rat hemorheology whole blood viscosity of table 10 meridians (x ± s)

Compare with the normal control group: * P＜0.05; * P＜0.01; Compare with model control group: △ P＜0.05; △ △ P＜0.01

7, the easypro antiinflammatory action of meridians

7.1 Oleum Tiglii is caused the effect of mice auricle swelling

Get 72 of mices, the male and female dual-purpose, body weight 18～22g, be divided at random the positive group of blank group, Western medicine (dexamethasone, subcutaneous injection), the positive group of Chinese medicine (TANGMAIKANG), meridians relax high dose group, meridians relax in dosage group, the meridians low dose group of relaxing.Every group 12.Begin administration after the grouping, every day 1 time, for three days on end, the blank group is given equivalent normal saline (20ml/kg).After the last administration 30 minutes with 2% Oleum Tiglii mixing proinflammatory agent (2% Oleum Tiglii, 20% dehydrated alcohol, 5% distilled water and 73% ether) 50 μ l are evenly coated in the mouse right ear exterior feature and cause inflammation, after causing scorching 4 hours mice is put to death, cut ears and lay round auricle at same position with the 6mm card punch, use scales/electronic balance weighing, the difference heavy with the left auricle in the right side is the swelling degree, estimates the influence of medicine to auricle inflammation due to the Oleum Tiglii.

The result shows that meridians relax mouse knoting oil is not seen that to the acute inflammatory reaction of auricle edema inhibitory action is arranged, and see Table 11.

The easypro influence to Fructus Crotonis oiliness inflammation of table 11 meridians (x ± s)

Group	Dosage (1/kg)	Number of animals	Auricle swelling degree (mg)
				The blank group		??12	??28.83±6.45
The dexamethasone matched group	??4mg	??12	??6.17±2.52 **
				The TANGMAIKANG matched group	??5g	??12	??26.92±6.72
The meridians high dose group of relaxing	The 4g crude drug	??12	??27.42±7.24
				Dosage group during meridians relax	The 2g crude drug	??12	??28.08±10.76
The meridians low dose group of relaxing	The 1g crude drug	??12	??28.75±7.72

Compare with the blank group: * * P＜0.01

7.2 to the swollen influence that forms of rat granuloma

Get 60 of Wistar rats, the male and female dual-purpose, body weight 200～250g, etherization, under aseptic condition, make thoracic incision, the sterilization cotton balls implantation both sides axillary region of 5mg is subcutaneous, and postoperative is divided into the blank group at random, the Western medicine positive is organized (dexamethasone, subcutaneous injection), Chinese medicine positive group (TANGMAIKANG), the easypro high dose group of meridians, the easypro middle dosage group of meridians, the easypro low dose group of meridians.Every group 10.Postoperative beginning in the 2nd day administration, the blank group is given equivalent normal saline (10ml/kg), every day 1 time, continuous 8 days, the 9th day execution rat, peel off and take out the granuloma induced by implantation of cotton pellets tissue, put into 60 ℃ of baking oven inner drying 12h, take out then and accurately weigh, deduct the raw cotton ball weight, be the granuloma net weight, estimate medicine to the outgrowth influence of granuloma induced by implantation of cotton pellets with this.

The result shows that the high dose group meridians relax to the obvious suppression effect that is formed with of granuloma induced by implantation of cotton pellets, see Table 12.

The easypro influence to the swollen formation of rat granuloma of table 12 meridians (x ± s)

Compare with the blank group: * P＜0.05

The analgesic activity that 8 meridians relax

8.1 hot plate method

Put the aluminum groove in 55 ± 0.5 ℃ of waters bath with thermostatic control, 1 of female mice is got in preheating 10 minutes at every turn, puts into groove, the record mice occur adding the metapedes required time (second/s) as this Mus pain threshold.Allly add that foot is less than 5s at the time or greater than 30s or the jump escapee is arranged, give it up.Get 72 of qualified mices, body weight 18～22g, be divided at random the positive group of blank group, Western medicine (morphine, lumbar injection), the positive group of Chinese medicine (TANGMAIKANG), meridians relax high dose group, meridians relax in dosage group, the meridians low dose group of relaxing, 12 every group.The administration respectively of each mice of grouping back, every day 1 time, for three days on end, the blank group is given equivalent normal saline (20ml/kg).Measure after the last administration 30,60 minutes pain threshold respectively.Each 1min that observes, as analgesia reaction in the 1min, pain threshold then calculated with 60 seconds.

The result shows, meridians relax after the high dose group administration 30,60min all can prolong mice and lick the sufficient time first, shows that meridians relax can improve the mice pain threshold, has significant analgesia role, sees Table 13.

The analgesic activity (hot plate method) that table 13 meridians relax (x ± s)

Group	Dosage (1/kg)	Number of animals	30min pain threshold (s) behind the medicine	60min pain threshold (s) behind the medicine
					The blank group		??12	?20.58±8.15	??23.42±7.19
The morphine matched group	??10mg	??12	?43.03±14.31 **	??44.23±14.22 **
					The TANGMAIKANG matched group	??5g	??12	?28.50±9.77 *	??28.71±6.92
The meridians high dose that relaxes	The 4g crude drug	??12	?28.92±10.16 *	??33.86±9.58 **
					Dosage during meridians relax	The 2g crude drug	??12	?24.08±8.88	??29.00±9.89
The meridians low dosage that relaxes	The 1g crude drug	??12	?21.92±11.48	??25.42±9.65

Compare with the blank group: * P＜0.05; * P＜0.01

8.2 writhing method

Get 72 of mices, the male and female dual-purpose, body weight 18～22g, be divided at random the positive group of blank group, Western medicine (morphine, lumbar injection), the positive group of Chinese medicine (TANGMAIKANG), meridians relax high dose group, meridians relax in dosage group, the meridians low dose group of relaxing, 12 every group.The administration respectively of each mice of grouping back, every day 1 time, for three days on end, the blank group is given equivalent normal saline (20ml/kg).After the last administration 30 minutes, each mouse peritoneal was injected 0.6% acetic acid 0.2ml, observed the writhing response number of times that mice occurs in 20 minutes behind injection acetic acid.

The result shows that the meridians of high dose group relax can obviously reduce the mouse writhing number of times, and showing has analgesic activity, sees Table 14.

The analgesic activity (writhing method) that table 14 meridians relax (x ± s)

Compare with the blank group: * P＜0.05; * P＜0.01

9, to the influence of rat capillary permeability

Get 50 of Wistar rats, the male and female dual-purpose, body weight 200～250g is divided into blank group, Western medicine positive controls (chlorphenamine at random, intramuscular injection), Chinese medicine positive controls (TANGMAIKANG), meridians relax high dose group, meridians relax in dosage group, the meridians low dose group of relaxing, 10 every group.The administration respectively of each rat of grouping back, every day 1 time, for three days on end, blank group is given equivalent normal saline (10ml/kg).After the last administration 30 minutes, at rat back subcutaneous injection 1ug/ml (0.1mg%) histamine 0.1ml, the 1% azovan blue normal saline 4ml/kg of intravenous injection immediately, put to death rat behind the 20min, cut the painted skin speckle in back, shredding and place the 5ml normal saline--acetone (3: 7) mixed liquor soaks 24h, and the centrifuging and taking supernatant is measured optical density value (0D) with 721 spectrophotometers in the 610nm place, with optical density value size reflection capillary permeability.

The result shows, relax height, middle dosage group of meridians has certain inhibitory action to the rat capillary permeability increase due to the histamine, and wherein the suppression ratio with high dose group, middle dosage group is respectively 21.1,9.14%, sees Table 15.

The easypro influence to the rat capillary permeability of table 15 meridians (x ± s)

Compare with the blank group: * P＜0.05

Above result comprises the peripheral nervous inflammation for the meridians relieving capsule is used for diabetic neuropathy, and diabetes autonomic nervous dysfunction provides good experimental basis.

Compared with prior art; Chinese medicine preparation of the present invention is a kind of new Chinese medicine for the treatment of the diabetes nerve complication; diabetes os hypogastroidale nerve had significant protective effect; can improve the sciatic conduction velocity of diabetes; malonaldehyde (MDA) and the sorbitol level that raises there is certain reduction effect; the activity effect of being significantly improved to glutathion peroxidase GSH-PX; to superoxide dismutase (SOD) activity that the descends effect that also improves; can obviously reduce blood red egg level of saccharifying and whole blood contrast viscosity; also has significant analgesia role; neural complication to diabetes has good therapeutic effect; for providing a kind of new medication, extensive patients selects; its raw material type is few; product quality is easy to control, and the preferred for preparation method of selecting through testing sieve provided by the present invention is simple; be easy to produce.

The specific embodiment

Embodiment 1: the preparation of capsule

Raw material: Radix Salviae Miltiorrhizae 612g, Radix Notoginseng 136g, silymarin 20g

Preparation method: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 6 times of amount 85% ethanol extractions 2 times, each 2 hours, filter merging filtrate, filtrate recycling ethanol, medicinal residues add 8 times of water gagings and decoct extraction 3 times, each 1 hour, filter, merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merges concentrated solution and ethanol extract ,-0.06MPa～-the 0.08MPa condition under microwave vacuum drying become dry extract, pulverize, add Radix Notoginseng fine powder and silymarin, add adjuvant, mixing, incapsulate, make 1000 of capsules.

Usage and dosage: oral, three times on the one, each 3～4.

Embodiment 2: the preparation of tablet

Raw material: Radix Salviae Miltiorrhizae 570g, Radix Notoginseng 160g, silymarin 30g

Preparation method: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 4 times of amount 95% ethanol extractions 2 times, each 3 hours, filter merging filtrate, filtrate recycling ethanol, medicinal residues add 12 times of water gagings and decoct extraction 2 times, each 0.5 hour, filter merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merges concentrated solution and ethanol extract, drying, pulverize, add Radix Notoginseng fine powder and silymarin, add adjuvant, mixing, tabletting is made 1000 in tablet.

Usage and dosage: oral, three times on the one, each 3～4.

Embodiment 3: the preparation of drop pill

Raw material: Radix Salviae Miltiorrhizae 640g, Radix Notoginseng 100g, silymarin 10g

Preparation method: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 8 times of amount 75% ethanol extractions 1 time, each 3 hours, filter, filtrate recycling ethanol, medicinal residues add 6 times of water gagings and decoct extraction 4 times, each 1.5 hours, filter, merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merge concentrated solution and ethanol extract, drying is pulverized, add Radix Notoginseng fine powder and silymarin and adjuvant, preparation process is made drop pill 1000 balls routinely.

Usage and dosage: oral, three times on the one, each 3～4 balls.

Embodiment 4: the preparation of capsule

Raw material: Radix Salviae Miltiorrhizae 660g, Radix Notoginseng 110g, silymarin 12g

Preparation method: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 7 times of amount 80% ethanol extractions 3 times, each 1 hour, filter merging filtrate, filtrate recycling ethanol, medicinal residues add 10 times of water gagings and decoct extraction 3 times, each 2 hours, filter, merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merges concentrated solution and ethanol extract, drying is pulverized, and adds Radix Notoginseng fine powder and silymarin, add adjuvant, incapsulate, make 1000 of capsules.

Usage and dosage: oral, three times on the one, each 3～4.

Embodiment 5: the preparation of granule

Raw material: Radix Salviae Miltiorrhizae 590g, Radix Notoginseng 170g, silymarin 27g

Preparation method: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 5 times of amount 90% ethanol extractions 3 times, each 3 hours, filter merging filtrate, filtrate recycling ethanol, medicinal residues add 9 times of water gagings and decoct extraction 4 times, each 0.5 hour, filter merging filtrate, being concentrated into 50 ℃ of relative densities is 1.05, merges concentrated solution and ethanol extract, drying, pulverize, add Radix Notoginseng fine powder and silymarin, granulate, drying, 1000 bags of granules are made in pack.

Usage and dosage: oral, three times on the one, each 3～4 bags.

Claims (6)

1. Chinese medicine preparation for the treatment of diabetic neuropathy, it is characterized in that: according to listed as parts by weight, this Chinese medicine preparation is to be prepared from for 10～30 parts by 570～660 parts of raw material of Chinese medicine Radix Salviae Miltiorrhizaes, 100～170 parts of Radix Notoginseng and silymarin.

2. according to the Chinese medicine preparation of the described treatment diabetic neuropathy of claim 1, it is characterized in that: according to listed as parts by weight, this Chinese medicine preparation is to be prepared from for 15～25 parts by 610～620 parts of raw material of Chinese medicine Radix Salviae Miltiorrhizaes, 125～145 parts of Radix Notoginseng and silymarin.

3. according to the Chinese medicine preparation of the described treatment diabetic neuropathy of claim 2, it is characterized in that: according to listed as parts by weight, this Chinese medicine preparation is to be prepared from for 20 parts by 612 parts of raw material of Chinese medicine Radix Salviae Miltiorrhizaes, 136 parts of Radix Notoginseng and silymarin.

4. the preparation method of the Chinese medicine preparation of the arbitrary described treatment diabetic neuropathy of claim 1-3 is characterized in that: gets Radix Notoginseng and pulverizes, and standby, get Radix Salviae Miltiorrhizae, add ethanol extraction, filter, filtrate recycling ethanol, the medicinal residues extracting in water filters, concentrate, merge concentrated solution and ethanol extract, drying is pulverized, add Radix Notoginseng fine powder and silymarin, preparation process is made various dosage forms routinely.

5. according to the preparation method of the Chinese medicine preparation of the described treatment diabetic neuropathy of claim 4, it is characterized in that: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 4～8 times of amount 60%～90% ethanol extractions 1～3 time, each 1～3 hour, filter merging filtrate, filtrate recycling ethanol, medicinal residues add 6～12 times of water gagings and decoct extraction 2～4 times, each 0.5～2 hour, filter merging filtrate, when being concentrated into 50 ℃ of relative densities is 1.05, merge concentrated solution and ethanol extract, drying is pulverized, add Radix Notoginseng fine powder and silymarin, preparation process is made various dosage forms routinely.

6. according to the preparation method of the Chinese medicine preparation of the described treatment diabetic neuropathy of claim 5, it is characterized in that: get Radix Notoginseng powder and be broken into fine powder, standby, get Radix Salviae Miltiorrhizae, add 6 times of amount 85% ethanol extractions 2 times, each 2 hours, filter, merging filtrate, filtrate recycling ethanol, medicinal residues add 8 times of water gagings and decoct extraction 3 times, each 1 hour, filter, merging filtrate is 1.05 when being concentrated into 50 ℃ of relative densities, merge concentrated solution and ethanol extract,-0.06MPa～-the 0.08MPa condition under microwave vacuum drying become dry extract, pulverize, add Radix Notoginseng fine powder and silymarin, add adjuvant, mixing incapsulates, and promptly gets capsule.