CN1202830C - Use of Rhizoma Dioscoreae Nipponicae and its dioscorea saponin in treating chronic glomerulus nephritis - Google Patents
Use of Rhizoma Dioscoreae Nipponicae and its dioscorea saponin in treating chronic glomerulus nephritis Download PDFInfo
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- CN1202830C CN1202830C CN 02159077 CN02159077A CN1202830C CN 1202830 C CN1202830 C CN 1202830C CN 02159077 CN02159077 CN 02159077 CN 02159077 A CN02159077 A CN 02159077A CN 1202830 C CN1202830 C CN 1202830C
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Abstract
The present invention relates to the new application of Japanese yam rhizome which is a traditional Chinese medicine and dioscorea total saponin which is an extract of the Japanese yam rhizome for curing chronic glomerulonephritis. The traditional Chinese medicine of Japanese yam rhizome and the dioscorea total saponin which is an extract of the Japanese yam rhizome have the functions of reducing urinary protein excretion, lowering high blood pressure and alleviating pathological damage to glomeruli, and therefore, the functions of treating chronic nephritis and improving renal functions are achieved.
Description
Invention field
The invention relates to Chinese medicine Rhizoma Dioscoreae Nipponicae and extract Rhizoma Dioscoreae total saponins thereof the new purposes in the treatment chronic glomerulonephritis, belong to the field of Chinese medicines.
Background technology
The Rhizoma Dioscoreae total saponins is the effective site of Chinese medicine Rhizoma Dioscoreae Nipponicae.Chemical constituent contains dioscin (dioscin), prolongs genus saponin multiple steroidal saponins such as (trillin).Dioscin content is 1.5～2.6%, still contain a small amount of 25-different-spiral shell steroid-3,5-diene (25-D-spirosta-3,5-diene) the Chinese medicine Rhizoma Dioscoreae Nipponicae is the dry rhizome of Dioscoreaceae plant Rhizoma Dioscoreae Nipponicae Rhizoma Dioscoreae (Dioscorea nipponica Makino), main product in northeast, North China, also there is distribution in areas such as Sichuan, Shaanxi, Gansu.Have relaxing muscles and tendons to promote blood circulation, the effect of the diuretic that helps digestion, the preventing the attack (or recurrence) of malaria that eliminates the phlegm.Clinical report Dioscorea nipponica Mak. Ningpo Yam Rhizome total saponins has certain curative effect to treatment coronary heart disease, atherosclerosis, blood fat reducing, relieving asthma antiinflammatory and antitumor.
Summary of the invention
The objective of the invention is to open Chinese medicine single Rhizoma Dioscoreae Nipponicae and extract Rhizoma Dioscoreae total saponins thereof the new purposes in the treatment chronic glomerulonephritis.
The effect in the treatment chronic glomerulonephritis of Chinese medicine Rhizoma Dioscoreae Nipponicae and extract Rhizoma Dioscoreae total saponins thereof is to be confirmed by following effect experiment, clinical experiment.
Chinese medicine Rhizoma Dioscoreae Nipponicae and its extract Rhizoma Dioscoreae total saponins have the urinary protein excretion of alleviating, reduce hypertension, alleviate the change of glomerule pathology, thereby reach the treatment chronic nephritis, improve the effect of renal function.
Experimental example 1Chinese medicine Rhizoma Dioscoreae Nipponicae and extract Rhizoma Dioscoreae total saponins thereof the drug effect in the treatment chronic glomerulonephritis
Experiment
One, materials and methods:
(1), the preparation of animal model
Male Wistar rat excised left kidney after 24 hours, and tail vein injection 500 μ g monoclonal antibody 1-22-3 cause chronic mesangial proliferative glomerulonephritis, observe altogether for 6 weeks after the modeling.
(2), the preparation of experimental drug: get prepared in laboratory Rhizoma Dioscoreae total saponins routinely.
(3), experimental design
In 7 ages in week of 28 male Wistar rats, divide four groups at random, 7 every group.The 1st group: compare for normal saline (PBS); The 2nd group, give Rhizoma Dioscoreae Nipponicae crude drug decoct 3g/ kg body weight/day.The 3rd group, give Rhizoma Dioscoreae total saponins 50mg/ kg body weight/day.The 4th group, give benazepril 2mg/ kg body weight/day.The antibody injection is gastric infusion simultaneously, observes altogether 42 days.Put to death whole rats on the 42nd day, heart extracting blood after winning right kidney and weighing, is got nephridial tissue and is done immunofluorescence and light microscopy checking.
(4), urine protein examination
Get the 1st, 7,14,21,28,35, the 42 day twenty-four-hour urine in monoclonal antibody 1-22-3 injection back, utilize the Bradford method, check 24 hours proteic content in the urine.
(5), rat blood pressure is measured
Before getting urine, check the blood pressure of rat, method be with rat be fixed on checkout gear (SoftronECGprocessor, Tokyo, Japan) in, heat to 30 ℃, survey the systolic pressure and the diastolic pressure of afterbody blood vessel, common triplicate is averaged.We have measured the variation of the 1st, 7,21,31 and 41 day blood pressure after the rat modeling.
(6), light microscopy
The part nephridial tissue is placed in 10% formalin solution fixing, and paraffin embedding is cut into the thick thin slice of 4 μ m with tissue, carries out PAM and PAS dyeing.We have observed 100 glomerule every tissue slice, and mesentery substrate hypertrophy in the glomerule has been carried out semi-quantitative analysis.And calculated the number of crescent in 100 glomerule.
(7), fluoroimmunoassay
The part nephridial tissue is chilled in rapidly in the N-ethane liquid, puts-70 ℃ of preservations.Freezing tissue is cut into 3-4 μ m thin slice, utilizes indirect immunofluorescence method dyeing.An employed antibody has: anti-transforming growth factor (TGF-β) antibody, anti-I type collagen antibodies and Chinese People's Anti-Japanese Military and Political College's Mus smooth muscle actin (antibody etc. of α-SM-actin).The anti-rabbit immunoglobulin Igs of the pig of FITC labelling is as the secondary antibodies of TGF-β and type i collagen; The goat anti-mouse igg 2a of FITC labelling is as the secondary antibodies of α-SM-actin.We have checked 20 glomerule in every tissue slice, according to the method for people such as Floege (Kidney Int.1992), have carried out semi-quantitative analysis.Promptly, can divide negative and positive according to glomerule fluorescence scope and intensity.It is negative that we are decided to be " 0 ".In the positive, the fluorescence scope is " 1+ " less than 25%; 25%～50% is " 2+ "; 50～75% is " 3+ "; Greater than 75% is " 4+ ".Calculate in every section the average of a glomerule then.With all tissue slices,, carry out statistical procedures by group categories.
(8), blood biochemical analysis
We have checked the content of respectively organizing rat plasma total protein, albumin, T-CHOL, serum creatinine and blood urea nitrogen.
(9), statistical procedures
Relatively adopt rank test and non-paired t test method between group.
Two. experimental result
(1) urine protein result:
Chinese medicine Rhizoma Dioscoreae Nipponicae and Dioscorea nipponica Mak. Ningpo Yam Rhizome total saponins suppressed urine protein on the 1st, 7,14,21,28,35 and 42 day significantly after modeling drainage.Compare statistical procedures with the normal saline matched group significant difference (seeing Table 1) is arranged.
Table 1.24 hour urine protein result (x+s)
1 day 7 days 14 days 21 days 28 days 35 days 42 days
Group n mg/d mg/d mg/d mg/d mg/d mg/d mg/d
PBS 7 15.04±2.94 76.84±25.43 83.97±24.53 70.50±13.95 54.69±15.34 45.27±7.85 35.47±6.51
Rhizoma Dioscoreae Nipponicae 3g 7 9.93 ± 1.96
* Δ Δ Δ46.66 ± 9.77
*45.96 ± 12.92
* *34.63 ± 4.18
* *30.29 ± 5.42
* * Δ24.63 ± 4.43
* *22.67 ± 6.51
*
Dioscin 50mg 7 13.96 ± 1.94 50.30 ± 5.36
*42.41 ± 3.19
* *37.84 ± 3.27
* *34.27 ± 4.33
*33.03 ± 4.27
* Δ Δ32.44 ± 4.90
Benazepril 4mg 7 15.79 ± 2.28 54.67 ± 5.61
*49.56 ± 3.51
*43.56 ± 5.39
* *34.27 ± 4.33
*37.59 ± 8.11 35.30 ± 5.34
Annotate: compare * P＜0.05, * * p＜0.001 with the PBS group; Compare with the benazepril group
ΔP＜0.05,
The Δ ΔP＜0.01
Δ Δ Δ,＜p＜0.001
(2) light microscopic and immunofluorescence result:
Chinese medicine Rhizoma Dioscoreae Nipponicae and Dioscorea nipponica Mak. Ningpo Yam Rhizome total saponins all can alleviate the pathological lesion of glomerule.Reduce the expansion of mesentery substrate and the formation (seeing Fig. 1, Fig. 2, Fig. 3) of crescent.Our semi-quantitative analysis Rhizoma Dioscoreae Nipponicae and Dioscorea nipponica Mak. Ningpo Yam Rhizome total saponins to transforming growth factor _ (TGF-β), (the influence of α-SM-actin) etc. of type i collagen (CollI) and rat smooth muscle actin, we find to have suppressed the expression in glomerule such as TGF-β, CollI and α-SM-actin significantly after the Drug therapy, and statistical procedures has the difference (seeing Table 2) of significance.
Table 2 immunofluorescence semi-quantitative analysis result (x ± s)
n TCF-β CollI α-SMA
PBS 7 1.06±0.30 1.05±0.34 1.13±0.26
Rhizoma Dioscoreae Nipponicae 3g 7 0.21 ± 0.13
* *0.18 ± 0.099
* * Δ0.40 ± 0.12
* *
Dioscin 50mg 7 0.32 ± 0.057
* *0.31 ± 0.073
* *0.46 ± 0.20
* *
Benazepril 4mg 7 0.37 ± 0.076
* *0.4 ± 0.082
* *0.53 ± 0.13
* *
Annotate: compare with the PBS group
* *P＜0.001; Compare with the benazepril group
ΔP＜0.05
(3) to the influence of rat blood pressure:
Shown in table 3,4, the blood pressure of rat increases gradually after the modeling, all showed hypertension at the 21st day, 31 days and 41 days, and Chinese medicine Rhizoma Dioscoreae Nipponicae and Dioscorea nipponica Mak. Ningpo Yam Rhizome total saponins all can make blood pressure reduce, with the depressor benazepril of generally acknowledging same effect (seeing Table 2,3) is arranged, learn by statistics and handle the difference that significance is arranged.
The situation of change of table 3 systolic pressure (x ± s)
1 day 7 days 21 days 31 days 41 days
Group n (mmHg) is (mmHg) (mmHg) (mmHg) (mmHg)
PBS 7 114.09±7.31 114.46±9.55 137.34±11.72 143.33±15.26 139.13±14.75
Rhizoma Dioscoreae Nipponicae 3g 7 113.33 ± 6.24
*109.3 ± 3.07 126.57 ± 4.56
*122.2 ± 3.77
* *123.36 ± 3.51
*
Dioscin 50mg 7 112.53 ± 6.88 116.07 ± 9.88 127.67 ± 4.18
*124.01 ± 4.02
* *123.64 ± 3.09
*
Benazepril 4mg 7 114.79 ± 4.76 115.01 ± 4.47 129.43 ± 3.29
*129.34 ± 2.71
*130.63 ± 5.33
Annotate: compare with the PBS group
*P＜0.05,
*P＜0.01,
* *P＜0.001
The situation of change of table 4 diastolic pressure (x ± s)
1 day 7 days 21 days 31 days 41 days
Group n (mmHg) is (mmHg) (mmHg) (mmHg) (mmHg)
PBS 7 82.77±6.74 81.39±8.66 94.50±7.30 101.00±11.94 99.39±14.24
Rhizoma Dioscoreae Nipponicae 3g 7 78.61 ± 4.45 77.76 ± 3.8 84.01 ± 3.32
*85.24 ± 4.83
* *83.27 ± 2.94
* *
Dioscin 50mg 7 86.84 ± 4.01 86.49 ± 4.4 87.46 ± 4.49
*86.06 ± 2.97
*85.31 ± 2.52
*
Benazepril 4mg 7 84.8 ± 3.18 87 ± 5.79 89.04 ± 3.70 90.51 ± 4.38
*89.07 ± 2.42
*
Annotate: compare with the PBS group
*P＜0.05,
*P＜0.01,
* *P＜0.001
(4) to the influence of blood biochemistry index:
As shown in table 5, Rhizoma Dioscoreae Nipponicae and Dioscorea nipponica Mak. Ningpo Yam Rhizome total saponins can improve the blood plasma total protein, improve renal function.
Table 5 blood biochemistry index (x ± s)
n T-protein T-cholest BUN Creatinine
(G/dl) (Mg/dl) (Mg/dl) (Mg/dl)
PBS 7 5.64±0.18 91.86±24.09 34.54±9.93 0.36±0.079
Rhizoma Dioscoreae Nipponicae 3g 7 5.61 ± 0.11 73.71 ± 9.07
*27.36 ± 4.09
*0.31 ± 0.069
Dioscin 50mg 7 6.04 ± 0.15
* * Δ Δ Δ83.14 ± 7.34 26.24 ± 6.26
*0.29 ± 0.069
Benazepril 4mg 7 5.66 ± 0.19 81.43 ± 12.20 31.93 ± 3.26 0.34 ± 0.053
Annotate: compare with the PBS group
*P＜0.05,
* *P＜0.001; Compare with the benazepril group
Δ Δ Δ＜P＜0.001
Experimental example 2The clinical experiment of Chinese medicine Rhizoma Dioscoreae Nipponicae in the treatment chronic glomerulonephritis
1 clinical data
1.1 case selects all clinical manifestations to meet the chronic nephritis diagnostic criteria, has got rid of the probability of secondary glomerulopathies such as acute postinfectious glomerulonephritis, anaphylactoid purpura, lupus nephritis.
1.2 physical data clinical observation 20 examples are the 1999-2001 of China-Japan Friendship Hospital and are in hospital or the out-patient.Wherein male 21 examples, women 9 examples; Age 16-62 year, average 37.53 years old; Course of disease 2-10.Clinical manifestation is that simple proteins is urinated 7 examples, albuminuria hematuria 7 examples of holding concurrently, nephrotic syndrome 6 examples.10 examples have the pathological diagnosis result, IgA nephropathy 2 examples wherein, non-IgA mesangial proliferative nephritis 5 examples, minute lesion 3 examples.
2 Therapeutic Method
The patient does not use any Western medicine, and the simple oral Rhizoma Dioscoreae Nipponicae is main Chinese herbs decoction, every day 2 times, about at every turn 100ml.Or the patient uses the Western medicine curative effect not obvious merely, and urine protein continues not disappear.In the time of the Western medicine decrement, cooperate and take the Chinese herbs decoction that Rhizoma Dioscoreae Nipponicae is the master.
3 criterions of therapeutical effect and therapeutic outcome
3.1 the efficacy assessment standard that criterion of therapeutical effect is passed through with reference to national nephropathy Symposium for the second time.Alleviate fully: urine protein continues negative, and urine erythrocyte disappears under the high power lens.The basic alleviation: urine protein continuous decrease 〉=50%, urine erythrocyte is no more than 4 under the high power lens.Take a turn for the better: urine protein continuous decrease 〉=25%, urine erythrocyte is no more than 4 under the high power lens.Invalid: urine protein or urine erythrocyte do not have significant change.
3.2 therapeutic outcome
In 20 examples, alleviate 3 examples (15%) fully, alleviate 6 examples (30%) substantially, 9 examples that take a turn for the better (45%), invalid 2 examples (10%), total effective rate is 90%.Curative effect obviously is better than the simple western medicine treatment.
Description of drawings:
Fig. 1 is a physiology saline control group;
Fig. 2 is a Rhizoma Dioscoreae total saponins treatment group;
Fig. 3 forms for the Rhizoma Dioscoreae Nipponicae treatment.
The specific embodiment
Embodiment 1
Precision takes by weighing Rhizoma Dioscoreae Nipponicae coarse powder 1000g, and decoction pieces is smashed, and crosses 20 mesh sieves, and add 6 times of water gagings and decoct 2 times, each 2 hours, merge decoction liquor and above-mentioned aqueous solution, filter, filtrate is concentrated in right amount, uses an amount of alcohol granulation, and drying, granulate, packing promptly get granule.Oral, a 0.25g, twice on the one.
Embodiment 2
Rhizoma Dioscoreae total saponins raw material 1000g, medical starch 1000g, mix homogeneously, the 0# capsule of packing into gets capsule.Every 0.35g, each oral 1-2 grain, twice of every day.
Embodiment 3
Rhizoma Dioscoreae total saponins raw material 1000g, medical starch 500g, mix homogeneously is used an amount of alcohol granulation, through the pelletizing machine granulate, tabletting, every 0.25g, oral, each 1-2 sheet, twice of every day.
Claims (4)
1, the application of single Rhizoma Dioscoreae Nipponicae in preparation treatment chronic glomerulonephritis medicine.
2, application as claimed in claim 1, wherein said treatment chronic glomerulonephritis are meant and alleviate urinary protein excretion, reduce hypertension, alleviate glomerule pathology infringement.
3, the application of Rhizoma Dioscoreae total saponins in preparation treatment chronic glomerulonephritis medicine.
4, the application described in claim 3, wherein said treatment chronic glomerulonephritis are meant and alleviate urinary protein excretion, reduce hypertension, alleviate glomerule pathology infringement.
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