CN103768305A - Medicine for treating spleen-deficiency diarrhea type IBS (Irritable Bowel Syndrome) and manufacturing method thereof - Google Patents

Abstract

The invention relates to a medicine for treating spleen-deficiency diarrhea type IBS (Irritable Bowel Syndrome), which uses Chinese medical herbs as the raw material, and a manufacturing method of the medicine. At present, when the spleen-deficiency diarrhea type IBS is treated by western medicine, no special therapy or drugs can be used, the symptomatic treatment manner is mainly adopted clinically, the untoward effects are various and side effects are larger after long-term use of western medicines, the possibility of relapse is high after drug withdrawal, and the long-term follow-up result is poor. According to the invention, Codonopsis pilosula, atractylodes macrocephala, poria cocos, radix paeoniae alba, Rhizoma Pinellinae Praeparata, Saposhnikovia divaricata, dried tangerine or orange peel, humifuse euphorbia herb, liquorice and cortex albiziae are weighed by proportion, and ground into powder, so as to prepare a conventional medicament for oral use. Traditional Chinese medicines are used for achieving the effect of treating both manifestation and root cause of disease, the total effective rate is 96%, radical treatment is implemented based on pathology; the effect is ideal, side effects are avoided, and pain of patients suffering from the spleen-deficiency diarrhea type IBS can be relieved.

Description

A kind of medicine and manufacture method thereof for the treatment of insufficiency of the spleen diarrhea irritable bowel syndrome

Technical field

The present invention relates to a kind of Chinese crude drug that utilizes and treat the medicine of insufficiency of the spleen diarrhea irritable bowel syndrome for raw material, also relate to the manufacture method of this medicine.

Background technology

Irritable bowel syndrome (irritable bowel syndrome, IBS) be the modal functional disease of gastrointestinal tract, also be a kind of biology-physiology-social disease, constitutional general chronic disease take smooth muscle function disorder as main manifestations, patient often has stomachache, abdominal distention, intestinal ring, diarrhoea, the symptoms such as constipation or diarrhoea and constipation replace, and with obvious general neurosis shape, the morphology and the biochemistry that lack soluble symptom are abnormal, it is the modal dysfunction of digestive tract, for digestive tract, syndrome due to the mental status and enteric cavity factor triple interaction.China's irritable bowel syndrome prevalence accounts for general population's 15%, accounts for 1/3～1/2 of Gastroenterology Clinic, and primary disease can be divided into diarrhea-type, constipation type, mixed type and typing Four types not, and diarrhea-type, for more common, accounts for 65% of irritable bowel syndrome.

Western medical treatment there is no specific short and medicine at present, mainly anti symptom treatment clinically, if to have loose bowels as main alone diarrhea, montmorillonite powder is common diarrhea, if take stomachache as main alone gastrointestinal antispasmodic medicine, Trimebutine Maleate capsule is the gastrointestinal smooth muscle motor adjustment agent that acts on potassium, calcium channel, has the dual regulation to gastrointestinal smooth muscle.Western medicine medicine has a lot, and treatment all exists limitation.And IBS sickness rate is high, and the course of disease is long, the many side effect of prolonged application Western medicine untoward reaction are larger, easily recurrence after drug withdrawal, and late result is not good.To thering is the patient of obvious mental symptom, suitably give tranquilizer, antidepressants, antianxiety drugs has certain help.Part diarrhea-type patient may have the disorder of intestinal microbial population, and application beneficial bacteria of intestinal tract class preparation is helpful.5-HT3 receptor antagonist alosetron is that master's IBS is effective to suffering from diarrhoea.

Summary of the invention

The present invention is directed to above-mentioned deficiency, a kind of insufficiency of the spleen diarrhea irritable bowel syndrome pathogeny is provided, utilizes Chinese Traditional Medicine treating both the principal and secondary aspects of a disease, total effective rate reaches 96%, utilize Chinese crude drug to treat the medicine of insufficiency of the spleen diarrhea irritable bowel syndrome for raw material, also relate to the manufacture method of this medicine.

Treat a medicine for insufficiency of the spleen diarrhea irritable bowel syndrome, it is made up of effective ingredient and adjuvant, and the ratio of weight and number of wherein making the raw material of effective ingredient is:

Radix Codonopsis 10～20, the Rhizoma Atractylodis Macrocephalae 10～20, Poria 10～20, the Radix Paeoniae Alba 15～30, Rhizoma Pinelliae Preparatum 5～15, Radix Saposhnikoviae 10～15, Pericarpium Citri Reticulatae 5～15, Herba Euphorbiae Humifusae 15～30, Radix Glycyrrhizae 4～10, Cortex Albiziae 15～20;

Treat a medicine for insufficiency of the spleen diarrhea irritable bowel syndrome, its ratio of weight and number of making the raw material of effective ingredient is:

Radix Codonopsis 15～20, the Rhizoma Atractylodis Macrocephalae 15～20, Poria 15～20, the Radix Paeoniae Alba 20～30, Rhizoma Pinelliae Preparatum 10～15, Radix Saposhnikoviae 10～12, Pericarpium Citri Reticulatae 5～10, Herba Euphorbiae Humifusae 15～20, Radix Glycyrrhizae 4～8, Cortex Albiziae 18～20.

Treat a medicine for insufficiency of the spleen diarrhea irritable bowel syndrome, its ratio of weight and number of making the raw material of effective ingredient is:

Radix Codonopsis 15, the Rhizoma Atractylodis Macrocephalae 15, Poria 15, the Radix Paeoniae Alba 20, Rhizoma Pinelliae Preparatum 10, Radix Saposhnikoviae 12, Pericarpium Citri Reticulatae 10, Herba Euphorbiae Humifusae 20, Radix Glycyrrhizae 5, Cortex Albiziae 20.

A kind of manufacture method of medicine for the treatment of insufficiency of the spleen diarrhea irritable bowel syndrome is:

Press drug ratio and weigh, break into powder, make conventional dose oral.

Radix Codonopsis replenishing QI to invigorate the spleen, the fang-feng powder liver spleen that relaxes, is monarch drug altogether; Poria eliminating dampness by diuresis spleen invigorating, the sweet warming middle-JIAO of Radix Glycyrrhizae Preparata, Pericarpium Citri Reticulatae circulation of qi promoting, invigorating the spleen for eliminating dampness, Rhizoma Pinelliae drying dampness to eliminate phlegm.Rhizoma Atractylodis Macrocephalae the spleen strengthening and damp drying, the eliminating pathogen in the liver of Radix Paeoniae Alba preserving YIN with astringents and nourishing blood is minister; Assistant is with the Rhizoma Pinelliae, Pericarpium Citri Reticulatae, Poria circulation of qi promoting, dampness, promoting diuresis with drugs of tasteless flavour, and Herba Euphorbiae Humifusae heat-clearing and toxic substances removing antidiarrheal, prevents warm-dryness syndrome heat-transformation, Cortex Albiziae resolving depression; With Radix Glycyrrhizae for making, during sweet temperature is adjusted.

Treatment by Chinese herbs is insufficiency of the spleen diarrhea irritable bowel syndrome, is than the maximum advantage of Western medicine, effects a radical cure from pathology.Effect is very desirable, is free from side effects, and can solve insufficiency of the spleen diarrhea irritable bowel syndrome patient's misery.Insufficiency of the spleen diarrhea irritable bowel syndrome is not one day and just, treatment also should be followed basic logic, adopt instant effect of the present invention, have no side effect, though flavour of a drug of the present invention are few, but make a distinction between the important and the lesser one, this mark is ruled together, and take originally as main, has embodied reason, method, the one line perforation of square medicine, combine the taste of a few taste medicines, its distinctive effect is strengthened in performance.

Accompanying drawing explanation

Below in conjunction with drawings and Examples, the present invention is further described:

Fig. 1 extraction process specificity of the present invention is investigated collection of illustrative plates

Note: 1. peoniflorin; 2. Hesperidin; A. peoniflorin reference substance; B. Hesperidin reference substance; C negative control (lacking Pericarpium Citri Reticulatae); D. negative control (lacking the Radix Paeoniae Alba); E the present invention

Fig. 2 peoniflorin standard curve of the present invention

Fig. 3 Hesperidin standard curve of the present invention

Its effect of clinical practice by the several years is remarkable, has carried out animal pharmacodynamic study, describes especially exemplified by several examples:

Radix Et Rhizoma Rhei is caused to the invigorating the spleen to arrest diarrhea effect of Mice with Spleen diarrhoea

Grouping: mice is divided into Normal group, model group, positive group and 12 administration groups (in A high dose, A in dosage, A low dosage, B high dose, B in dosage, B low dosage, C high dose, C in dosage, C low dosage, D high dose, D dosage, D low dosage) according to table of random number, wherein ABCD represents respectively soup decocting liquid of the present invention, water soluble part, n-butanol portion, ethyl acetate extract, 10 every group.After grouping, record every the weight of animals, normally raise.

Modeling: except Normal group, all the other are respectively organized gavage and give Radix Et Rhizoma Rhei decocting liquid, concentration 1g medical material/L, only, continuous 8 days, Normal group gave same volume distilled water to administration volume 1mL/.After last gavage Radix Et Rhizoma Rhei decocting liquid, record each group of Mouse Weight and loose stool rate.

Administration: modeling finishes to start for latter second day, respectively organizes continuous gavage medicine 7 days, and Normal group and model group give same volume distilled water, and administration volume is 0.2mL/10g.

Index determining: again record each group of Mouse Weight and loose stool rate before the last administration of each treated animal, after record, fasting be can't help water 24 hours, 30 min gavage prepared Chinese ink (prepared Chinese ink: water: black sesame paste=1mL:3mL:0.4g after last administration, 0.5mL/ only), after 20min, mice is put to death in cervical region dislocation, dissect rapidly and take out intestinal, record the distance (pylorus advances the distance of front end to prepared Chinese ink) that small intestinal total length and prepared Chinese ink advance in intestinal with ruler, calculate propelling rate (propelling rate=advance distance/total length).Take out thymus and spleen simultaneously, weigh, calculate organ coefficient.

2.2 impacts of soup of the present invention on insufficiency of the spleen diarrhea irritable bowel syndrome rat

Grouping: rat is divided into Normal group, model group, positive group and 12 administration groups (in A high dose, A in dosage, A low dosage, B high dose, B in dosage, B low dosage, C high dose, C in dosage, C low dosage, D high dose, D dosage, D low dosage) according to table of random number, wherein ABCD represents respectively soup decocting liquid of the present invention, water soluble part, n-butanol portion, ethyl acetate extract, 10 every group.After grouping, record every the weight of animals, measure the intestinal sensitivity of every animal under normal circumstances, normally raise.

Modeling: modeling process is divided into three phases, first week: gavage Folium Sennae decocting liquid (0.3g crude drug/L, 20mL/kg); Second week: continue gavage Folium Sennae decocting liquid, after having gavaged, fetter crop, preepipodite, chest with adhesive tape, the restriction preepipodite Head And Face of scratching, but do not limit its activity, cause certain stress stimulation, constraint times 2 h/ days; The 3rd week: only fetter crop, preepipodite, chest with adhesive tape, the restriction preepipodite Head And Face of scratching, but do not limit its activity, cause certain stress stimulation, constraint times 2 h/ days.

Administration: from modeling the 3rd week, be that gavage gives relative medicine after unmuzzling, Normal group and model group give distilled water, and continuous 7 days, administration volume was 10mL/kg.

Index determining: modeling, after 2 weeks, is recorded each treated animal body weight and intestinal sensitivity.Intestinal sensitivity testing method: test rat fasting the previous day be can't help water 24 hours, one people fixes rat head, another people inserts catheter along rat anus, the about 5cm of the degree of depth, conduit and rat tail root tangle up, after 15min, treat that rat conforms with rubber band, with 2.5mL syringe gradually to injecting distilled water expansion intestinal in catheter, observe and cause the volume threshold that rat abdomen lifts and arch upward in back respectively, carry out behavior evaluation.For obtaining assessment result accurately, after 15min, each threshold value is all repeated to expansion, totally 3 times, data are got average.

Before last administration, then record as stated above body weight and the intestinal sensitivity of every animal.Half an hour after last administration, gavage prepared Chinese ink (prepared Chinese ink: water: black sesame paste=1mL:3mL:0.4g, 2.5mL/ only), puts to death after 20min, measures pylorus and advances length foremost to prepared Chinese ink, calculates propelling rate (propelling rate=advance distance/total length).Then Pou Qu colon, 4 ℃ of normal saline flushings are clean, take 0.5g, make 10% homogenate with normal saline, after the centrifugal 10min of 1000g, get supernatant ELISA method and measure 5-HT.

2.3 soup Dichlorodiphenyl Acetates of the present invention cause the analgesic activity of mouse writhing reaction

Grouping: mice is divided into Normal group, positive group and 12 administration groups (in A high dose, A in dosage, A low dosage, B high dose, B in dosage, B low dosage, C high dose, C in dosage, C low dosage, D high dose, D dosage, D low dosage) according to table of random number, wherein ABCD represents respectively soup decocting liquid of the present invention, water soluble part, n-butanol portion, ethyl acetate extract, 10 every group.After grouping, record every the weight of animals, normally raise.

Administration and index determining: each group gives relative medicine or distilled water, continuous 8 days, 1h after last administration, every Mus lumbar injection 0.6% glacial acetic acid (0.2mL/ is only), records incubation period and the interior writhing number of times of 10min that writhing response appears in mice, relatively group difference.

3, experimental result

3.1 pairs of Radix Et Rhizoma Rhei cause the invigorating the spleen to arrest diarrhea effect of Mice with Spleen diarrhoea

3.1.1 Mouse Weight changes

The impact of table 1 soup of the present invention on mice with spleen deficiency body weight change (g,

± s)

Group	Animal number of elements	Before modeling	After modeling	After administration
					Normal group	12	23.3±1.6	25.3±1.0	24.6±1.2
Model group	12	23.6±0.9	22.6±0.6	22.7±0.6
					Positive group	12	23.7±1.1	23.9±1.8	23.5±0.9
A high dose	12	23.3±2.2	23.6±1.1	24.0±1.1
					Dosage in A	12	23.6±1.0	23.9±1.0	24.4±1.1 *
A low dosage	12	23.7±1.3	23.5±1.4	22.3±1.4
					B high dose	12	23.6±1.2	24.0±0.5	22.9±2.0
Dosage in B	12	23.2±1.2	23.7±1.8	22.2±0.9
					B low dosage	12	22.7±1.7	23.8±2.2	22.5±2.1
C high dose	12	23.4±1.2	23.7±1.7	22.5±1.5
					Dosage in C	12	23.3±0.9	23.2±2.3	22.1±2.1
C low dosage	12	23.3±1.0	23.1±2.3	20.2±2.3
					D high dose	12	23.2±1.5	22.9±1.2	22.4±1.4
Dosage in D	12	23.0±0.9	23.3±0.7	25.8±1.8 *
					D low dosage	12	23.4±1.0	24.1±1.5	23.6±1.6

Note: with model control group comparison, ^*represent p< 0.05, ^*represent psame under < 0.01().

From table 1: before modeling, body weight no significant difference between each treated animal; Through continuous 8 days gavage Radix Et Rhizoma Rhei decoctings, Normal group Mouse Weight normal growth, every of average about 2.0g, and all the other each group weight of mice are slow, all lower than Normal group; After administration 7 days, with model group comparison, in A in dosage, D dosage group body weight gain obviously ( p< 0.05), all the other each tested medicine group body weight gains have no significant change.

3.1.2 mice loose stool rate

The impact of table 2 soup of the present invention on mice with spleen deficiency loose stool rate (

± s)

Group	Animal number of elements	Loose stool rate %
			Normal group	12	5.66±1.22
Model group	12	36.51±5.41
			Positive group	12	28.24±2.30
A high dose	12	9.09±0.89 *
			Dosage in A	12	7.50±0.66 *
A low dosage	12	21.15±1.43
			B high dose	12	27.27±3.08
Dosage in B	12	12.50±2.42 *
			B low dosage	12	10.69±3.03 *
C high dose	12	38.89±4.25
			Dosage in C	12	17.14±4.50
C low dosage	12	16.06±3.07
			D high dose	12	12.90±1.49 *
Dosage in D	12	27.24±2.46
			D low dosage	12	30.00±2.00

From table 2: model group mice loose stool rate is 36%, apparently higher than Normal group ( p<0.05), in the agent of tested medicine A senior middle school, B, low dose, D high dose can obviously reduce respectively insufficiency of the spleen diarrhea mice loose stool rate, and wherein in A, dosage antidiarrheal effect is best.

3.1.3 mice intestinal advances

Table 3 soup of the present invention on mice with spleen deficiency intestinal advance impact (cm,

± s)

Group	Animal number of elements	Propelling rate
			Normal group	12	81.7±5.9
Model group	12	86.9±9.4
			Positive group	12	82.8±6.2 *
A high dose	12	75.9±7.1 *
			Dosage in A	12	97.8±1.6
A low dosage	12	66.9±29.3 *
			B high dose	12	89.5±13.0
Dosage in B	12	89.0±2.8
			B low dosage	12	96.0±4.1
C high dose	12	87.5±2.2
			Dosage in C	12	86.7±5.2
C low dosage	12	95.4±4.4
			D high dose	12	85.5±12.2
Dosage in D	12	88.4±11.9
			D low dosage	12	91.6±1.7

From table 3: compare with Normal group, model group mice intestinal advance distance is elongated, and propelling rate obviously increases, may be due to insufficiency of the spleen diarrhoea; Positive drug, A high dose, A low dosage all obviously reduce the intestinal advance distance of mice with spleen deficiency and propelling rate ( p<0.05), improve the motor function of intestinal.

3.1.4 mouse thymus and spleen gland index

The impact of table 4 soup of the present invention on insufficiency of the spleen Thymus and spleen index (× 100,

± s)

Group	Animal number of elements	Thymus index	Index and spleen index
				Normal group	12	0.2105±0.0782	0.2476±0.0566
Model group	12	0.2539±0.0734	0.2418±0.0276
				Positive group	12	0.2094±0.0300	0.2337±0.0635
A high dose	12	0.2764±0.0250	0.2525±0.0312
				Dosage in A	12	0.3214±0.1395	0.2126±0.0563
A low dosage	12	0.3114±0.1535	0.3961±0.2302 *
				B high dose	12	0.1984±0.0373	0.2383±0.0497
Dosage in B	12	0.3004±0.0106	0.2185±0.0222
				B low dosage	12	0.2443±0.0962	0.2347±0.0439
C high dose	12	0.2692±0.1169	0.2779±0.0854
				Dosage in C	12	0.2862±0.0465	0.3392±0.2365
C low dosage	12	0.1817±0.0986	0.2601±0.1008
				D high dose	12	0.1000±0.0063	0.1682±0.0547
Dosage in D	12	0.2023±0.0314	0.2887±0.0132
				D low dosage	12	0.2499±0.0707	0.2950±0.0890

From table 4: the Mice with Spleen Diarrhea Model due to Radix Et Rhizoma Rhei, recover voluntarily after one week, model group Thymus and Spleen index recovers normal level.Each administration group is compared with model group, and A, B have rising trend to thymus index, and A, C, D have rising trend to index and spleen index, and therefore, tested medicine different parts may have respectively the effect that promotes mice with spleen deficiency thymus and spleen index to return to normal level.

3.1.5 brief summary

Comprehensive, soup decocting liquid of the present invention can raise mice with spleen deficiency body weight due to Radix Et Rhizoma Rhei, to insufficiency of the spleen diarrhoea there is anti-diarrhea effect, can improve mice with spleen deficiency intestinal movement dysfunction, can there is the effect that promotes mice with spleen deficiency thymus and spleen index to return to normal level; Soup water soluble part of the present invention has anti-diarrhea effect to insufficiency of the spleen Diarrhea Model mice; Ethyl acetate extract has the effect of rising mice with spleen deficiency body weight, antidiarrheal; And causing mice with spleen deficiency to Radix Et Rhizoma Rhei, n-butanol portion substantially do not have a significant effect.

3.2 impacts on insufficiency of the spleen diarrhea irritable bowel syndrome rat

3.2.1 rat body weight changes

Impact that table 5 soup of the present invention changes insufficiency of the spleen diarrhea irritable bowel syndrome rat body weight (g,

± s)

Group	Animal number of elements	Before modeling	After modeling	After administration
					Normal group	10	201.3±9.9	209.3±22.7	219.3±28.3
Model group	10	196.3±6.1	183.6±23.6	197.4±21.6
					Positive group	10	190.0±5.6	171.6±24.8	185.0±24.6
A high dose	10	201.9±8.0	189.8±14.3	206.7±15.6
					Dosage in A	10	191.0±6.7	177.7±15.0	191.0±19.2
A low dosage	10	193.7±12.2	194.2±30.0	209.0±37.9
					B high dose	10	194.9±10.0	190.7±15.8	211.0±20.4 *
Dosage in B	10	192.2±7.4	188.6±26.7	206.5±33.7
					B low dosage	10	205.4±7.6	193.4±12.2	210.5±22.0 *
C high dose	10	194.7±7.7	181.7±8.4	193.5±9.4
					Dosage in C	10	202.7±11.8	183.3±10.5	198.7±12.2
C low dosage	10	205.7±10.7	196.4±18.0	204.5±22.4
					D high dose	10	193.3±12.4	200.0±28.2	209.2±29.2
Dosage in D	10	194.2±3.4	188.5±15.3	206.2±22.7
					D low dosage	10	193.1±10.6	192.3±20.4	206.2±27.3

From table 5: before modeling, body weight no significant difference between each treated animal; After modeling 2 weeks, rats in normal control group Normal-weight increases, 2 weeks every of about 8.0g of balanced growth, and all the other each group rat body weights become negative growth, and be starkly lower than Normal group; After administration 1 week, compare with model group, the high agent of B and low dosage can obviously promote rat body weight recovery ( p<0.05), A, B, D have the trend that promotes that rat body weight recovers, and C has no significant effect this rat model body weight change.

3.2.2 intestine in rats Susceptible change

The impact of table 6 soup of the present invention on insufficiency of the spleen diarrhea irritable bowel syndrome intestine in rats Susceptible change (L,

± s)

Group	Animal number of elements	Before modeling	After modeling	After administration
					Normal group	10	0.65±0.23	0.68±0.26	0.65±0.13
Model group	10	0.64±0.16	0.56±0.21	0.58±0.11
					Positive group	10	0.62±0.13	0.56±0.10	0.51±0.09
A high dose	10	0.64±0.17	0.56±0.20	0.59±0.14
					Dosage in A	10	0.66±0.18	0.62±0.17	0.55±0.14
A low dosage	10	0.62±0.16	0.48±0.20	0.52±0.11
					B high dose	10	0.54±0.09	0.51±0.11	0.52±0.12
Dosage in B	10	0.62±0.16	0.60±0.06	0.54±0.12
					B low dosage	10	0.60±0.14	0.61±0.19	0.52±0.10
C high dose	10	0.64±0.18	0.59±0.17	0.69±0.10 *
					Dosage in C	10	0.58±0.13	0.50±0.17	0.59±0.12
C low dosage	10	0.66±0.38	0.51±0.24	0.56±0.12
					D high dose	10	0.68±0.11	0.58±0.18	0.61±0.17
Dosage in D	10	0.60±0.29	0.59±0.13	0.55±0.09
					D low dosage	10	0.64±0.18	0.63±0.24	0.52±0.11

From table 6: before modeling, the threshold value no significant difference that between each treated animal, abdominal part lifts; After modeling 2 weeks, except Normal group, all the other each treated animal intestinal sensitivity threshold values are all lower than original level and Normal group, and most group reduces obvious with Normal group; After drug treatment, C high dose group intestinal sensitivity threshold value and the obvious increase of model group ( p<0.05), in A high dose group, C, dosage group, D high dose group intestinal sensitivity threshold value are also higher than model group, but no significant difference, A low dose group, B high dose group, C low dose group intestinal sensitivity threshold value higher than treatment before, all the other tested medicine group intestinal sensitivity threshold values lower than the administration of this group before and model group.

3.2.3 rat intestine advances

Impact that table 7 soup of the present invention advances insufficiency of the spleen diarrhea irritable bowel syndrome rat intestine (cm,

± s)

Group	Animal number of elements	Propelling rate
			Normal group	10	60.01±17.83
Model group	10	95.73±9.54
			Positive group	10	70.10±7.08 *
A high dose	10	94.00±13.42
			Dosage in A	10	100.00±0.00
A low dosage	10	81.07±15.68
			B high dose	10	73.82±16.74
Dosage in B	10	74.32±8.01 *
			B low dosage	10	73.22±17.86
C high dose	10	77.04±15.87
			Dosage in C	10	73.02±15.14
C low dosage	10	69.80±16.13 *
			D high dose	10	63.49±7.60 *
Dosage in D	10	67.27±5.51 *
			D low dosage	10	75.62±16.62

From table 7: compare with Normal group, model group rat intestine advance distance is elongated, and propelling rate obviously increases, may be due to insufficiency of the spleen diarrhoea, to cause Mice with Spleen experimental result consistent with Radix Et Rhizoma Rhei; After administration, dosage in B, C low dosage, the obvious reduction compared with model group of D senior middle school dosage group alvine pushing rate ( p<0.05), all the other respectively organize alvine pushing rate also lower than model group, but no significant difference.

3.2.4 rat colon is organized 5-HT level

The impact of diarrhea irritable bowel syndrome rat colon 5-HT level that table 8 soup of the present invention is insufficiency of the spleen (μ g/L,

± s)

Group	Animal number of elements	5-HT level
			Normal group	10	0.665±0.133
Model group	10	0.973±0.141
			Positive group	10	0.826±0.254
A high dose	10	0.899±0.147
			Dosage in A	10	0.792±0.190
A low dosage	10	0.818±0.224
			B high dose	10	0.934±0.202
Dosage in B	10	0.925±0.317
			B low dosage	10	0.894±0.178
C high dose	10	0.706±0.193 *
			Dosage in C	10	0.733±0.210 *
C low dosage	10	0.834±0.131
			D high dose	10	0.902±0.310
Dosage in D	10	0.885±0.141
			D low dosage	10	0.867±0.251

From table 8: model group and Normal group comparison, colon's 5-HT content obviously raises, administration group and model group comparison, high, the middle dosage group of the tested medicine C 5-HT of colon content obviously reduce ( p<0.05), all the other each group also has reduction trend.

3.2.5 brief summary

Comprehensive, to insufficiency of the spleen diarrhea irritable bowel syndrome rat, soup water soluble part of the present invention can raise its body weight, reduce the alvine pushing rate of rat, thereby it is abnormal to improve diarrhea irritable bowel syndrome intestine in rats motor function; Soup n-butanol portion of the present invention can reduce insufficiency of the spleen diarrhea irritable bowel syndrome intestine in rats sensitivity, reduces the alvine pushing rate of rat, reduce the 5-HT of colon content; Ethyl acetate extract can reduce the alvine pushing rate of rat, improves diarrhea irritable bowel syndrome intestine in rats motor function abnormal.

3.3 Dichlorodiphenyl Acetates cause the analgesic activity of mouse writhing reaction

The impact of table 9 soup of the present invention on mice incubation period and writhing number of times (

± s)

Group	Animal number of elements	Incubation period (second)	Writhing number of times
				Normal group	12	230.00±60.81	14.33±1.15
Positive group	12	265.00±19.80	10.50±2.12 *
				A high dose	12	313.33±21.15 *	14.67±1.12
Dosage in A	12	104.60±10.54	18.60±6.71
				A low dosage	12	187.50±57.28	21.50±0.71
B high dose	12	194.60±25.83	20.00±2.74
				Dosage in B	12	226.75±42.17	15.00±5.41
B low dosage	12	256.50±107.75	14.25±4.99
				C high dose	12	311.20±106.23	12.60±2.02
Dosage in C	12	257.50±78.62	13.00±6.73
				C low dosage	12	238.33±25.97	16.00±5.17
D high dose	12	277.40±80.18	12.60±3.91
				Dosage in D	12	269.67±95.01	11.00±1.64 *
D low dosage	12	215.20±8.79	16.20±6.14

Note: with Normal group comparison, ^* p<0.05, ^* p<0.01.

From table 9: A high dose group compared with Normal group, obviously extend incubation period ( p<0.05); Normal group is compared, in positive group and D dosage group writhing number of times obviously reduce ( p<0.05).

4, sum up

Comprehensive, soup decocting liquid of the present invention can raise mice with spleen deficiency body weight due to Radix Et Rhizoma Rhei, to insufficiency of the spleen diarrhoea there is anti-diarrhea effect, can improve mice with spleen deficiency intestinal movement dysfunction, may there is the effect that promotes mice with spleen deficiency thymus and spleen index to return to normal level; Soup water soluble part of the present invention has anti-diarrhea effect to insufficiency of the spleen Diarrhea Model mice; Ethyl acetate extract has the effect of rising mice with spleen deficiency body weight, antidiarrheal; And causing mice with spleen deficiency to Radix Et Rhizoma Rhei, n-butanol portion substantially do not have a significant effect.To insufficiency of the spleen diarrhea irritable bowel syndrome rat, soup water soluble part of the present invention can raise its body weight, reduce the alvine pushing rate of rat, thereby it is abnormal to improve diarrhea irritable bowel syndrome intestine in rats motor function; Soup n-butanol portion of the present invention can reduce insufficiency of the spleen diarrhea irritable bowel syndrome intestine in rats sensitivity, reduces the alvine pushing rate of rat, reduce the 5-HT of colon content; Ethyl acetate extract can reduce the alvine pushing rate of rat, improves diarrhea irritable bowel syndrome intestine in rats motor function abnormal.

Study on extraction of the present invention

Show that from the research of preliminary experiment peoniflorin, Hesperidin may be its main material bases, in order to control its quality, this section is investigated the extraction process of decocting liquid of the present invention, for the quality control that this compound recipe is further developed as after hospital preparation is established certain basis.

Also carried out Study on extraction, spy is exemplified below:

1 instrument and reagent

1.1 instrument

High performance liquid chromatograph (Shimadzu LC-20AT), DAD detector; BP121S type electronic balance (German Sartorius company); BUG25-12 ultrasonic cleaner (must believe ultrasonic company limited); Chromatographic column: Diamonsil diamond C18 (250 mm × 4.6 mm, 5 μ are m); Excellent general UPT series Superpure water machine (Chengdu You Pu Electronic Products Inc.).

1.2 reagent

Reference substance: peoniflorin (lot number: 110736-200526), Hesperidin (lot number: 0721-9909) are all purchased from Nat'l Pharmaceutical & Biological Products Control Institute, for assay.

Reagent: chromatograph acetonitrile and chromatograph methanol (Fisher Scientific), ultra-pure water, phosphoric acid are analytical pure (Chengdu Ke Long chemical reagent factory);

Medical material: the Radix Paeoniae Alba (lot number: 1002055), the Rhizoma Atractylodis Macrocephalae (lot number: 0907034), Radix Ginseng Rubra (lot number: 0910138), Poria (lot number 1003209), Radix Glycyrrhizae (lot number: 0905126), Pericarpium Citri Reticulatae (lot number: 0910010), the Rhizoma Pinelliae (lot number: 0902003), Herba Euphorbiae Humifusae (lot number 0803008 :), Radix Saposhnikoviae (lot number: 1006118), 1002113) etc. Cortex Albiziae (lot number: the prepared slices of Chinese crude drugs are all purchased from Sichuan new Flos Nelumbinis prepared slices of Chinese crude drugs limited company, identified by associate professor Long Fei of TCD identification teaching and research room of Chengdu University of Traditional Chinese Medicine, all meet 2010 version " Chinese Pharmacopoeia " requirement.

2 methods and result

2.1 peoniflorins, content of hesperidin are measured

2.1.1 chromatographic condition

Shimadzu LC-20AT high performance liquid chromatograph, DAD detector; Chromatographic column: Diamonsil diamond C18 (250 mm × 4.6 mm, 5 μ m), mobile phase: acetonitrile: 0.1% phosphoric acid water=20:80; Flow velocity: 1.0ml/min; Column temperature: 35 ℃; Detect wavelength: 230nm.

2.1.2 the preparation of reference substance solution

Respectively precision weighing peoniflorin, Hesperidin are appropriate, are mixed with concentration and are respectively the reference substance of 0.1021mg/ml, 0.0961mg/ml with methanol, for subsequent use.

2.1.3 the preparation of need testing solution

Take the decoction pieces of prescription 1/10 amount, require to extract 9 samples according to orthogonal design, filter, filtrate volatilizes under 80 ℃ of water-baths, and drying under reduced pressure 24h gets the about 1.0g of dry extract, accurately weighed, add 25ml methanol, weigh, ultrasonic 1h, taking-up is put dry rear benefit and is weighed, filtration, precision measures subsequent filtrate 0.2ml, in the volumetric flask of 1ml, cross 0.45 μ m microporous filter membrane by methanol constant volume, for subsequent use.

2.1.4 the preparation of negative sample

Prepare respectively containing the Radix Paeoniae Alba with not containing the negative sample of Pericarpium Citri Reticulatae decoction pieces according to prescription, be prepared into corresponding negative sample according to the preparation method under 2.1.3 item, for subsequent use.

2.2 methodological study

2.2.1 specificity experiment

Get reference substance solution, do not contain the Radix Paeoniae Alba and not containing negative sample and the need testing solution of Pericarpium Citri Reticulatae, respectively enter 10 μ L by condition under 2.1.1 item.Negative sample does not go out peak in the corresponding retention time of reference substance, illustrates to method specificity stronger.

seefig. 1,2,3,

2.2.2 linear relationship is investigated

Accurate peoniflorin reference substance 4,8,12,16, the 20 μ L that draw under 2.1.2 item, with the chromatographic condition sample introduction under 2.1.1 item, measure peak area.Take peoniflorin reference substance X as independent variable, peak area Y is dependent variable, peoniflorin standard curve Y=5 × 10 ⁹x-92957 (r=0.9999); Same method, Hesperidin sample size is 4,8,12,16,20 μ L, obtaining Hesperidin standard curve is Y=2 × 10 ⁹x-9768 (r=0.9999).Show that peoniflorin, Hesperidin are respectively at 0.4048-2.014 μ g, 0.3844-1.922 μ g, linear relationship is good.

2.2.3 precision test

Reference substance solution 10 μ L under accurate absorption 2.1.2 item respectively, continuous sample introduction 6 times, O.20% the RSD that calculates peoniflorin, Hesperidin peak area is respectively, and 0.22%, O.28%.Show that precision is good.

2.2.4 replica test

Get 6 parts of same test samples, press respectively 2.1.3 item below legal system standby, with the 10 μ L of chromatographic condition sample introduction under 2.1.1 item, result peoniflorin average quality mark is 9.0103mg/g, and RSD O.58%; Hesperidin average quality mark is 6.5527mg/g, and RSD O.58%.Show that this law repeatability is good.

2.2.5 stability test

Get same test sample, standby by 2.1.3 item below legal system, with chromatographic condition under 2.1.1 item respectively 0,2,4,6,8,10h sample introduction 10 μ L, result peoniflorin RSD is O.58%; Hesperidin RSD 1.2%.Show that test sample is stable in 12h.

2.2.6 application of sample recovery test

Get same extracting mode (No:4: add the water of 10 times of amounts, decoct 2 times, each 30min.Peoniflorin, Hesperidin mass fraction are about respectively 9.000mg/g, the test sample of different content 3.300mg/g) is appropriate, concrete content sees the following form, accurately weighed, totally 3 parts, precision adds reference substance (to be respectively 80% of test sample content respectively, 100%, 120%), by 3 parts of the each preparations of the sample of the each different content of method under 2.1.3 item, totally 9 parts.With the 10 μ L of chromatographic condition sample introduction under 2.1.1 item, measure the content of peoniflorin, Hesperidin.Result shows, the accuracy of this law is better.

Table 10 extraction process average recovery of the present invention

2.3 sample determination

Get 3 batch samples, 2 parts every batch, by 2.1.3 item below legal system available test sample solution, measure by chromatographic condition sample introduction under 2.1.1 item, the results are shown in Table 3.

Table 11 extraction process peoniflorin of the present invention content of hesperidin

2.4 yield of extract are measured

Precision measures the each 10ml of above subsequent filtrate, is placed in the evaporating dish that is dried to constant weight, after 80 ℃ of water-baths volatilize, in 105 ℃ of dry 3h of baking oven, places in exsiccator and is cooled to room temperature, accurately weighed immediately, calculates paste-forming rate.The results are shown in Table 11.

2.5 Orthogonal Experiment and Design

2.5.1 determining of factor level

This prescription adopts decocting to carry, and amount of water (A), decocting time (B), 3 factors of decoction number of times (C) is investigated as index take peoniflorin, Hesperidin, paste-forming rate, uses L ₉(3 ⁴) orthogonal table carries out optimal process, in table 4.

The design of table 12 decocting technological factor level

Level	A amount of water/doubly	B decocting time/min	C decoction number of times/time
				1	5	30	1
2	10	60	2
				2	15	90	3

2.5.2 evaluation criterion

In this compound recipe, the Radix Paeoniae Alba is monarch drug, and Pericarpium Citri Reticulatae is ministerial drug, carries out aggregative weighted scoring therefore be respectively 0.5,0.3,0.2 weight coefficient according to peoniflorin, Hesperidin, paste-forming rate.Orthogonal test is in table 13, and variance analysis is in table 14.

Table 13 water extraction extraction process orthogonal test

NO	A	B	C	D	Peoniflorin mg/g	Hesperidin mg/g	Paste-forming rate %	Comprehensive grading
									1	1	1	1	1	9.5905	3.2170	19.35	9.630
2	1	2	2	2	7.1160	3.0066	27.61	9.982
									3	1	3	3	3	5.3234	4.3330	47.07	13.38
4	2	1	2	3	5.7997	2.8412	37.51	11.25
									5	2	2	3	1	3.7616	3.6263	45.06	11.98
6	2	3	1	2	6.1390	2.5219	29.63	9.752
									7	3	1	3	2	4.3736	3.9703	42.05	11.79
8	3	2	1	3	9.5555	3.1234	30.47	11.81
									9	3	3	2	1	3.6715	3.3397	42.91	11.42
K1	10.997	10.890	10.397	11.010
									K2	10.994	11.257	10.884	10.508
K3	11.673	11.517	12.383	12.147
									R	0.679	0.627	1.986	1.639

Comprehensive grading=peoniflorin × 0.5 ﹢ Hesperidin × 0.3+ paste-forming rate × 0.2

The variance analysis of table 14 orthogonal test comprehensive grading

Soruces of variation	SS	f	F	P
					A	0.918	2	0.302	>0.05
B	0.596	2	0.196	>0.05
					C	6.429	2	2.113	>0.05

Result: can find out from table 13 data extreme difference, the each factor effect primary and secondary that affects extraction effect is C>A>B, and the optimum extraction process obtaining is A ₃b ₃c ₃.Variance estimation shows, A, and B, C factor there are no significant difference, from producing, the angle of energy savings, time is considered, selected A ₂b ₂c ₂for optimum extraction process, add 10 times of water gagings and extract 2 times, each lh.

2.6 process certification tests

Take 3 parts of the decoction pieces of prescription 1/10 amount, extract according to preferred extraction process condition, mensuration peoniflorin, Hesperidin, paste-forming rate, the results are shown in Table 15. results and show that selection process is stable, reasonable, feasible.

Table 15 demonstration test result

3 brief summaries and discussion

The peoniflorin, Hesperidin and the paste-forming rate that change take compatibility front and back, as index, have been investigated extraction process of the present invention.Optimised process extracts 2 times for adding 10 times of water gagings, each lh.Experimental results show that process stabilizing, reasonable, feasible.

Problem adopts L ₉(3 ⁴) orthogonal table carries out optimal process, result shows 9 kinds of extracting method, the variation of peoniflorin, Hesperidin and paste-forming rate does not have significant difference.The mode that water extraction is described is little on the impact of our chemical composition content.

The specific embodiment

Embodiment mono-

Weigh:

Radix Codonopsis 15kg, Rhizoma Atractylodis Macrocephalae 15kg, Poria 15kg, Radix Paeoniae Alba 20kg, Rhizoma Pinelliae Preparatum 10 kg, Radix Saposhnikoviae 12 kg, Pericarpium Citri Reticulatae 10 kg, Herba Euphorbiae Humifusae 20 kg, Radix Glycyrrhizae 5 kg, Cortex Albiziae 20 kg.Break into powder, dry, mix, in filled capsules, make capsule.

Embodiment bis-

A, weighing: Radix Codonopsis 20kg, Rhizoma Atractylodis Macrocephalae 20kg, Poria 20kg, Radix Paeoniae Alba 25kg, Rhizoma Pinelliae Preparatum 15 kg, Radix Saposhnikoviae 15kg, Pericarpium Citri Reticulatae 15 kg, Herba Euphorbiae Humifusae 30 kg, Radix Glycyrrhizae 10kg, Cortex Albiziae 20 kg.Break into powder, decoct with water 2～4 times, collecting decoction, filters, and filtrate is 1.2～1.3 thick paste while being condensed into 80 ℃ of relative densities.

B, add the thick paste of 100～500 portions of Icing Sugar of A step raw material weight ratio of weight and number and A step to mix, granulation, the dry granule that to obtain.

Embodiment tri-

A, weighing: Radix Codonopsis 10kg, Rhizoma Atractylodis Macrocephalae 10kg, Poria 10kg, Radix Paeoniae Alba 15kg, Rhizoma Pinelliae Preparatum 5 kg, Radix Saposhnikoviae 10kg, Pericarpium Citri Reticulatae 5 kg, Herba Euphorbiae Humifusae 15 kg, Radix Glycyrrhizae 4kg, Cortex Albiziae 15 kg.Break into powder, filter, filtrate is 1.0～1.6 thick paste while being condensed into 80 ℃ of relative densities;

B, the thick paste of 1～2 part of starch and A step is mixed, granulation, dry;

C, the dry granule in B step is pressed into tablet, sugar coating.

Claims (4)

1. treat a medicine for insufficiency of the spleen diarrhea irritable bowel syndrome, it is characterized in that it is made up of effective ingredient and adjuvant, the ratio of weight and number of wherein making the raw material of effective ingredient is:

Radix Codonopsis 10～20, the Rhizoma Atractylodis Macrocephalae 10～20, Poria 10～20, the Radix Paeoniae Alba 15～30, Rhizoma Pinelliae Preparatum 5～15, Radix Saposhnikoviae 10～15, Pericarpium Citri Reticulatae 5～15, Herba Euphorbiae Humifusae 15～30, Radix Glycyrrhizae 4～10, Cortex Albiziae 15～20.

2. treat a medicine for insufficiency of the spleen diarrhea irritable bowel syndrome, its ratio of weight and number of making the raw material of effective ingredient is:

Radix Codonopsis 15～20, the Rhizoma Atractylodis Macrocephalae 15～20, Poria 15～20, the Radix Paeoniae Alba 20～30, Rhizoma Pinelliae Preparatum 10～15, Radix Saposhnikoviae 10～12, Pericarpium Citri Reticulatae 5～10, Herba Euphorbiae Humifusae 15～20, Radix Glycyrrhizae 4～8, Cortex Albiziae 18～20.

3. treat a medicine for insufficiency of the spleen diarrhea irritable bowel syndrome, its ratio of weight and number of making the raw material of effective ingredient is:

Radix Codonopsis 15, the Rhizoma Atractylodis Macrocephalae 15, Poria 15, the Radix Paeoniae Alba 20, Rhizoma Pinelliae Preparatum 10, Radix Saposhnikoviae 12, Pericarpium Citri Reticulatae 10, Herba Euphorbiae Humifusae 20, Radix Glycyrrhizae 5, Cortex Albiziae 20.

4. the manufacture method of a kind of medicine for the treatment of insufficiency of the spleen diarrhea irritable bowel syndrome as described in claim 1,2 or 3 is:

Press drug ratio and weigh, break into powder, make conventional dose oral.

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