CN104027510A - Application of epation in preparation of medicines for preventing and treating xerophthalmia - Google Patents

Abstract

The invention discloses application of epation in preparation of medicines preventing and treating xerophthalmia. The epation has good effect in preventing and treating xerophthalmia, can be used for obviously increasing secretion of tear and improving the stability of tear film and cellular structure of ocular surface, and has a remarkable curative effect to the xerophthalmia-tear gland secretion function damage and corneal epithelium deficiency and partial inflammatory response caused by decrease of androgen, thereby having the effect similar to androgen.

Description

ZHANGYANMING PIAN is prevented and treated the application in the medicine of xerophthalmia in preparation

Technical field

The present invention relates to field of medicaments, particularly relate to ZHANGYANMING PIAN and prevent and treat the application in the medicine of xerophthalmia in preparation.

Background technology

Xerophthalmia (Keratoconjunctivitis Sicca or dry eye) refers to that the tear film stability that tear quality and quantity is abnormal or kinetics causes extremely that any reason causes declines, and causes a general name for the various diseases that table organization's pathological changes is feature.Clinical manifestation has eye sensation of dryness, foreign body sensation, burn feeling, gargalesthesia, photophobia, furious, blurred vision, vision fluctuation and asthenopia etc.Xerophthalmia is called again keratoconjunctivitis sicca, and motherland's medical science is referred to as dry astringent eye, xerosis conjunctivitis, belongs to " dry disease " category, is clinical common disease.Xerophthalmia is to be caused by the dynamic (dynamical) abnormal and abnormal institute of eye superficial epithelium of tear.Owing to being closely related between eye superficial epithelium and tear film, interdepend and do as a whole playing a role, rather than isolated exist, so xerophthalmia causes by many reasons, its cause of disease is very complicated.Various constitutional lacrimal gland diseases, sarcoid, graft versus host disease, HIV, lachrymal gland blocks, constitutional Sjogren syndrome, Secondary cases Sjogren syndrome, rheumatic arthritis, systemic lupus erythematosus (sle), systemic sclerosis, sand holes, burn, paralysis trace class sky lump skin ulcer, neuroparalytic keratitis, facial paralysis, meibomian gland dysfunction, blepharitis, mucin lacks, twinkle abnormal, margo palpebrae malposition, angle conjunctival operation otch, ultraviolet, contact lens is worn improper and some drugs (as tricyclic antidepressant, anti-handkerchief Jin Shi medicine, oral contraceptive, some resisting hypertension and anti-arrhythmic) all can cause that xerophthalmia occurs.

The primary categories of xerophthalmia is water liquid shortage type xerophthalmia (ADDE) and evaporated strong type xerophthalmia (EDE).ADDE is the exhaustion due to the lacrimal secretion of lachrymal gland, and this classification can further be further divided into xerodermosteosis xerophthalmia (by a self-immunprocess, for example rheumatoid arthritis targeting lachrymal gland and salivary gland) and non-xerodermosteosis xerophthalmia (Tear function obstacle, but except the systemic autoimmune feature of xerodermosteosis, age related xerophthalmia for example).EDE owing under normal tear fluid secretory function exists from the eye surface losses that exposes too much moisture.Its reason can be endogenous (for example, due to inherent sickness influence eyelid structure or kinetics, meibomian gland dysfunction) or exogen (wherein the generation of diseases of eye surface exposes owing to some exogen, for example vitamin A deficiency).

Xerophthalmia is a modal ocular disease, according to prevalence rate, estimates, 85% city work crowd suffers from xerophthalmia, and at present, increasing white collar personage is in vision " subhealth state " state.Because xerophthalmia can be caused by many reasons, its cause of disease is very complicated, so xerophthalmia morbidity crowd is very extensive; To old people, all likely cause xerophthalmia from child, youth, middle age, wherein child, youth mostly are environment or medicine factor causes, and middle-aged and elderly people mostly is physiology or pathological factor causes, and its total Therapeutic Principle is according to cause of disease individual comprehensive therapy.In preclinical therapy, external eye drop, ointment or the gel preparations of adopting are treated more, but external eye drop only has certain curative effect to the xerophthalmia of environment or the initiation of medicine factor, to the xerophthalmia of the physiology of middle-aged and elderly people or pathological factor initiation, can only alleviate its symptom, can not effect a permanent cure; And oral medicinal herb treatment adopts determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs, giving consideration to both the incidental and fundamental has certain advantage to the treatment of primary disease.

ZHANGYANMING PIAN is the exclusive product that Community in Baiyunshan, Guangzhou Zhong Yi pharmaceutcal corporation, Ltd produces, by Rhizoma Acori Graminei, Semen Cassiae, Herba Cistanches, Radix Puerariae, Semen Celosiae, Radix Codonopsis, Fructus Viticis, Fructus Lycii, Semen Plantaginis, the Radix Paeoniae Alba, Fructus Corni, Radix Glycyrrhizae, Semen Cuscutae, Rhizoma Cimicifugae, Nux Prinsepiae, Flos Chrysanthemi, Flos Buddlejae, Rhizoma Chuanxiong, stir-baked RHIZOMA POLYGONATI with yellow rice wine, Radix Rehmanniae Preparata, the 22 taste crude drug such as Cortex Phellodendri and the Radix Astragali are made, record in < < Chinese Pharmacopoeia > > 2010 editions, there is liver and kidney tonifying, during spleen invigorating is adjusted, yang invigorating profit key, the merit of removing nebula to improve visual acuity, be used for the treatment of at present the initial stage and mid-term senile cataract.

Summary of the invention

Based on this, the object of the present invention is to provide the new application of ZHANGYANMING PIAN.

The concrete technical scheme solving the problems of the technologies described above is as follows:

ZHANGYANMING PIAN is prevented and treated the application in the medicine of xerophthalmia in preparation.

In some embodiment, described xerophthalmia is video terminal xerophthalmia, mid-aged population xerophthalmia, cataract characteristic of disease xerophthalmia, diabetic xerophthalmia or the unbalance xerophthalmia causing of gonadal hormone therein.

In some embodiment, the dosage form of described medicine is tablet, capsule, granule, pill, oral liquid, external preparation or injection therein.

The new application of ZHANGYANMING PIAN of the present invention has the following advantages and beneficial effect:

The present invention draws through inventor's large quantity research and experiment: ZHANGYANMING PIAN has good action effect to prevention and treatment xerophthalmia, especially its can significantly increase tear secretion, improve tear film stability and eye table cellularity, to the xerophthalmia due to androgen levels decline, be also lacrimal secretion functional lesion and corneal epithelium disappearance and local inflammation reaction, have significant therapeutic effect, this shows that it has the effect of class androgenic.

Accompanying drawing explanation

Fig. 1 is Normal group and ZHANGYANMING PIAN group goblet cell density result block diagram in test 1;

Fig. 2 is goblet cell density and the aspect graph of Normal group and ZHANGYANMING PIAN group in test 1, wherein, and the goblet cell density that A is Normal group and aspect graph; B is goblet cell density and the aspect graph before the treatment of ZHANGYANMING PIAN group; C is ZHANGYANMING PIAN group treatment goblet cell density and the aspect graph of 20 days; D is ZHANGYANMING PIAN group treatment goblet cell density and the aspect graph of 30 days.

The specific embodiment

Below with reference to specific embodiment, the present invention will be further described.

Test the impact of 1 ZHANGYANMING PIAN on xerophthalmia

One, experiment purpose

This test, by making new zealand white rabbit xerophthalmia model, is observed the impact of ZHANGYANMING PIAN on rabbit xerophthalmia model lacrimal secretion, tear film stability and Oculofacial structure, inquires into the preventive and therapeutic effect of ZHANGYANMING PIAN to xerophthalmia.

Two, test material and method

2.1 laboratory animals and grouping

Healthy new zealand white rabbit, male and female are not limit, body weight 2.0～2.5kg.The raising of animal and environment are all followed zooperal standard in international ophthalmology and vision science research.Get 36 new zealand white rabbits and be divided into 3 groups by table of random number method: blank group, experiment contrast group, ZHANGYANMING PIAN group, 12 every group.

2.2 experimental animal models are made

It is that experimental eye drips by 0.1% Benzalkonii Chloridum (BAC) 4 weeks that all animals are chosen right eye, 2 times/d, set up xerophthalmia animal model (specifically referring to Burstein NL.Preservative cytotoxic threshold for benzal-konium chloride and Chlorhexidine digluconate in cat and rabbit cro-neas[J] .Invest Ophthalmol Vis Sci, 1980, 19:308-313), after 4 weeks, check dry eye model rabbit, see that angle conjunctiva outward appearance is dry, tear amount is few, the equal < 10mm/5min of Schirmer test value (the equal > 10mm of matched group), breakup time of tear film (BUT) is < 10s all, fluorescent staining is shown in angle conjunctiva point, slice colouring, is modelling success.

2.3 main reagent and instruments

ZHANGYANMING PIAN is that Community in Baiyunshan, Guangzhou Zhong Yi pharmaceutcal corporation, Ltd produces (batch number P00042), before experiment, pulverizes, and crosses 100 mesh sieves, and distilled water is mixed with concentration 0.2g/ml oral administration mixed suspension; Benzalkonii Chloridum (BAC, Sigma company product) tri-distilled water is mixed with concentration 0.1% solution; Schirmer tests filter paper bar (Tianjin Jingming New Technological Development Co., Ltd.); Fluorescein sodium ophthalmology Test paper (Tianjin Jingming New Technological Development Co., Ltd.); Nitrocellulose filter (PALL); Periodic acid-Schiff (PAS) staining kit (Yuan Ye bio tech ltd, Shanghai).Kica paraffin slicing machine (German Leica company produce); Hitachih-600 type transmission electron microscope (HIT's production).

2.4 medication

ZHANGYANMING PIAN oral administration mixed suspension, concentration 0.2g/ml, with front shaking up.Medication therapy groups gives the intervention of ZHANGYANMING PIAN gavage setting up on the basis of animal model, according to rabbit body weight proportion (1ml/kg) gavage every day 1 time; Experiment contrast group is gavage equal-volume normal saline simultaneously; Blank group is not done any treatment, and its normal eyes is as Normal group.

2.5 specimen samplings and detection

Laboratory animal checks by same people.Review time, place, brightness of illumination and temperature are identical.After administration the 10th, 20,30d observes respectively rabbit angle conjunctiva outward appearance, fluorescent staining, BUT under slit lamp; Do Schirmer test, conjunctiva impression cytology checks (CIC) simultaneously.30d after administration, puts to death after animal with air tap inserting method, gets at random 6 new zealand rabbit clip corneas and apart from limbus of corneae 2mm top center bulbar conjunctiva tissue, tissues observed pathological change under light microscopic for every group.

2.5.1Schirmer test

After Oxybuprocaine topical anesthesia, after 1min, cotton swab dips in xerophthalmia eyelid surrounding liquid, and Schirmer reagent paper is placed in to conjunctival sac, outer 1/3 intersection in rabbit below, remainder overhangs skin surface, close one's eyes, after 5min, take out filter paper bar, survey the wetted length of filter paper bar.

2.5.2 breakup time of tear film (BUT)

Fluorescein sodium ophthalmology test strip is placed on to lower tarsal conjunctiva capsule for a moment, fluorescein sodium is uniformly distributed in maintain eyelid after eye table to open, under slit lamp microscope, with cobalt blue light, observe, until there is the 1st dry speckle on tear film, record opens up into from eyelid the time that the 1st dry speckle occurs, continuous measurement three times, averages and record.

2.5.3 cornea fluorescent staining

Glass rod dips in fluorescein and is applied in lower tarsal conjunctiva capsule, the painted situation of viewing angle conjunctiva under naked eyes and slit lamp.

2.5.4 conjunctiva impression cytology checks (CIC)

Treatment after the 10th, 20,30d respectively organizes row conjunctiva marking cytolgical examination respectively.Nitrocellulose filter is cut into 3.5mm * 3.5mm size, with distilled water immersion 4h, dries, standby.After topical anesthesia, respectively two nitrocellulose filter matsurfaces are placed in downwards on nose and temporo on quadrantal spheres conjunctiva, continue to press to print for 10 seconds and get top layer epithelial cell.Filter membrane is placed in to 95% ethanol fixing.Periodic acid Schiff's reagent (PAS) dyeing.Under optical microscope, count goblet cell, according to Nelson standards of grading [specifically referring to: Nelson JD.Impression cytology[J] .Cornea, 1988,7:71-81] carry out classification.

2.5.5 histopathological examination

The Cornea and conjunctiva tissue of cutting is placed in to fixedly 24h of 10% formalin.After gradient dehydration, paraffin embedding and section, HE dyeing, conjunctiva adds does PAS dyeing.

2.6 statistical method

Adopt SPSS16.0 statistics software to carry out statistical analysis, experimental result with mean ± standard deviation ( ) represent.Same time point, each organizes Schirmer test value, BUT value more all adopts one factor analysis of variance method, and further adopts LSD-t method of inspection to compare between two.Difference in more same group between different time points, adopts paired t-test.The P < 0.05 of usining has statistical significance as difference.

Three, result of the test

3.1Schirmer test

Result is referring to table 1, as known from Table 1: the equal < 10mm of rabbit xerophthalmia model Schirmer test value.Before treatment, the Schirmer test value of blank group, experiment contrast group and ZHANGYANMING PIAN group is respectively (8.42 ± 0.83) mm, (8.58 ± 0.79) mm, (8.47 ± 0.73) mm, difference not statistically significant (P > 0.05).Treat 20d, ZHANGYANMING PIAN group Schirmer test value is (13.19 ± 0.67) mm, compared with obviously raise before medication (P < 0.01), compared with blank group and experiment contrast group, also obviously raise (P < 0.01), difference not statistically significant between blank group and experiment contrast group (P > 0.05); After medication 30d, ZHANGYANMING PIAN group Schirmertest value is (16.32 ± 1.43) mm, before treatment, obviously raise (P < 0.01), compared with blank group and experiment contrast group, also obviously raise (P < 0.01), difference not statistically significant between blank group and experiment contrast group (P > 0.05).

Table 1 is respectively organized the variation (mm) of Schirmer test value after medication

Note: compare * P < 0.05, * * P < 0.01 with same administration time blank group Schirmer test value.

3.2 breakup time of tear film (BUT)

Result is referring to table 2, as known from Table 2: the equal < 10s of BUT value of blank group, experiment contrast group and ZHANGYANMING PIAN group before treatment, be respectively (8.86 ± 1.31) s, (8.72 ± 0.75) s and (8.37 ± 1.13) s, difference not statistically significant (P > 0.05).Treat 20d, ZHANGYANMING PIAN group BUT value is (12.85 ± 1.75) s, before treatment, obviously extend (P < 0.01), compared with blank group and experiment contrast group, also obviously extend (P < 0.05), difference not statistically significant between blank group and experiment contrast group (P > 0.05); After medication 30d, ZHANGYANMING PIAN group BUT value is (14.52 ± 1.78) s, before treatment, obviously extend (P < 0.01), compared with blank group and experiment contrast group, also obviously extend (P < 0.001), difference not statistically significant between blank group and experiment contrast group (P > 0.05).

Table 2 is respectively organized the variation (s) of BUT value after medication

Note: compare * P < 0.05, * * P < 0.01 with same administration time blank group BUT value.

3.3 fluorescein sodium dyeing (FL)

After dry eye model completes, the visible rabbit of naked eyes angle conjunctiva outward appearance is dry matt; Fluorescent staining is shown in angle conjunctiva point, slice colouring.After ZHANGYANMING PIAN group medication 10d, rabbit angle conjunctiva outward appearance is dry compared with slightly taking a favorable turn before medication, and tear amount is few, but angle conjunctiva point, slice colouring are clearly better compared with xerophthalmia group: after treatment 20d, angle conjunctival xerosis further takes a turn for the better, and tear amount increases, and angle conjunctiva point, slice colouring are few; After treatment 30d, angle conjunctiva outward appearance is moistening, and tear obviously increases, and angle conjunctiva has no painted.

3.4 conjunctiva impression cytologies check (CIC)

Result is referring to Fig. 1, as can be seen from Figure 1: between dry eye model group and Normal group, the difference of goblet cell density has statistical significance.After administration, ZHANGYANMING PIAN group goblet cell density increases gradually, and before administration, compares difference and has statistical significance; Difference not statistically significant before each time point goblet cell density of blank group and experiment contrast group and administration.

The inspection of conjunctiva impression cytology adopts generally acknowledged Nelson standards of grading at present to carry out classification.Squamous metaplasia is divided into 4 ranks, 0 grade: normal; 1 grade: slight; 2 grades: moderate; 3 grades: severe.Normal lagophthalmos Nelson is classified as 0 grade, and xerophthalmia eye is classified as 2 grades.After administration, ZHANGYANMING PIAN group goblet cell density increases gradually, and it is normal that form is tending towards.No significant difference before each time point goblet cell density of blank group and experiment contrast group and form and administration, concrete outcome is referring to Fig. 2, (A) normal lagophthalmos in Fig. 2, Nelson is classified as 0 grade; (B) before treatment, Nelson is classified as 2 grades; (C) treatment is after 20 days, and Nelson is classified as 1 grade; (D) treatment is after 30 days, and Nelson is classified as 0 grade.(×200)。

3.5 light microscopy checking

Under microscope, can see, normal conjunctiva epithelium consists of epithelium layer and the 3-4 layer cuboid cell of column, and the goblet cell of the PAS positive interts wherein.In blank group and experiment contrast group, conjunctiva attenuation, cuboid cell layer disappears, and the goblet cell of the PAS positive significantly reduces.ZHANGYANMING PIAN group has no notable difference with normal lagophthalmos contrast Conjunctival Goblet Cells quantity, but conjunctival epithelium number of plies compared with normal lagophthalmos reduces.

Four, conclusion

Known from the above results: ZHANGYANMING PIAN group medication 20d, Schirmer value and BUT value all obviously increase (P < 0.01) compared with other two groups; Conjunctival cells trace is learned and is checked that discovery ZHANGYANMING PIAN group Conjunctival Goblet Cells quantity is compared with other two groups of showed increased; Light microscopy checking discovery ZHANGYANMING PIAN group is compared with other two groups, and the part conjunctival epithelium number of plies increases, goblet cell increases.

Conclusion: ZHANGYANMING PIAN group can obviously increase rabbit xerophthalmia model lacrimal secretion, improve tear film stability and eye table cellularity.

Test effect and the impact of 2 ZHANGYANMING PIAN on the male Mus xerophthalmia model of castration

One, test objective

This research, by making the male Mus xerophthalmia model of castration, is observed ZHANGYANMING PIAN and xerophthalmia is respectively organized to the variation of cornea and lachrymal gland light microscopic and lachrymal gland Electronic Speculum ultra micro organizational structure; And by ZHANGYANMING PIAN, it is intervened, observe the effect of ZHANGYANMING PIAN to the male Mus xerophthalmia model cornea of castration and lacrimal tissue, inquire into the action effect that it prevents and treats the made xerophthalmia of castration.

Two, test material and method

2.1 laboratory animals and grouping

Choose 150 1 monthly age of health male Wistar whitewash mouses (body weight 90～110g), be divided at random totally 15 groups of A1, B1, Cl, Dl, El, A3, B3, C3, D3, E3, A5, B5, C5, D5, E5,10 every group.Be respectively: A: normal group; B: Sham-operated control group; C: operating comparison (castration) group; D: androgen in treating group contrast; E: ZHANGYANMING PIAN treatment group; L representative was raised after 1 month; 3 for raising after 3 months: 5 for raising after 5 months.

2.2 experimental animal models are made

The male Mus xerophthalmia model of castration is made, with reference to related documents Ma Yiqun. the research [J] of castrated male rabbits xerophthalmia model corneal epithelial cell apoptosis and related gene expression. ophthalmology research, 2004,22 (3): 286-289.}, excises bilateral testes and epididymis by experimental mouse; Sham operated rats is only cut scrotum and Testectomy not; Normal group is left intact.

2.3 main reagent and instruments

ZHANGYANMING PIAN is that Community in Baiyunshan, Guangzhou Zhong Yi pharmaceutcal corporation, Ltd produces (batch number P00042), before experiment, pulverizes, and crosses 100 mesh sieves, and distilled water is mixed with concentration 0.2g/ml oral administration mixed suspension; Testosterone Propionate injection (25mg/ml, commercially available Tianjin Jin Yao aminoacid company limited produces); Kica paraffin slicing machine (German Leica company produce); Hitachih-600 type transmission electron microscope (HIT's production).

2.4 medication

Each group after modeling 1 week starts medication after wound heals substantially.Androgen in treating group Testosterone Propionate injection is injected with the rear 0.5ml/kg leg muscle of pressing of 1:4 Oleum Sesami dilution, 1 time on the every 3rd; ZHANGYANMING PIAN treatment group is with ZHANGYANMING PIAN 0.2g/ml oral administration mixed suspension 5ml/kg gavage, and every day, gavage was 1 time; All the other 3 groups with normal saline 5ml/kg gavage, and every day, gavage was 1 time.

2.5 specimen samplings and detection

Laboratory animal has checked by same people.Each review time, place, brightness of illumination, humidity and temperature are identical.After administration, each treated animal is respectively at l, after 3,5 months, and under slit lamp, viewing angle conjunctiva outward appearance, breakup time of tear film (BUT), cornea fluorescent staining check and Schimer test respectively.Administration l, after 3,5 months, each group is randomly drawed 10 rats, and after 20% urethane general anesthesia, head-breaking is put to death each treated animal, at once wins eyes lachrymal gland, cornea tissue, and 4% paraformaldehyde is fixing to be prepared, HE dyeing, each organizational structure of om observation.

2.5.1 cornea fluorescent staining

Glass rod dips in fluorescein and is applied in lower tarsal conjunctiva capsule, under slit lamp, observes, and cornea is divided into 4 quadrants, O level: dye-free (-), l level: be dispersed in point-like dyeing (+), 2 grades: intensive point-like dyeing (++), 3 grades: lamellar dyeing (+++).

2.5.2Schirmer test

After Oxybuprocaine topical anesthesia, after 1min, cotton swab dips in xerophthalmia eyelid surrounding liquid, with 2.5mm * 35mm tears detecting filter strip, by one end bending 5mm, is placed in inside lower eyelid in l/3 conjunctival sac, and remainder overhangs skin surface.After closing one's eyes 5 minutes, measure filter paper wetted length (not comprising opisthotonos).

2.5.3 breakup time of tear film (BUT)

With Glass rod, dip in 2% fluorescein sodium smears in lower tarsal conjunctiva capsule.Catacleisis fluorescein is uniformly distributed in after anterior corneal surface.Being fixed thereon lower eyelid fully exposes cornea.Under slit lamp, by cobalt blue rayed, observe, from the beginning timing of twinkling for the last time.Record the time that occurs first breakdown point on tear film.

2.5.4 histopathological examination

The cornea of cutting and lacrimal tissue are placed in to fixedly 24h of 10% formalin.After gradient dehydration, paraffin embedding and section, HE dyeing, conjunctiva adds does PAS dyeing.

2.6 statistical procedures

All data are all processed through SPSSl5.0 systems soft ware.Measurement data with average ± standard deviation ( ) represent.If meet normality and homogeneity of variance. use multifactor analysis of variance tournament method; While not meeting normality and homogeneity of variance.Use non-ginseng multiple comparisons method.The P < 0.05 of take has statistical significance as difference.

Three, result of the test

3.1 cornea fluorescent stainings

Result is referring to table 3, as known from Table 3: A, B group cornea fluorescent staining is fed after 3 months without obvious change: C group rat, and it is damaged that corneal epithelium starts carrying out property, and fluorescent staining is positive, and increases the weight of in time; D, E group are shown in cornea fluorescent staining in castration after 3,5 months, and stain reduces compared with C group.

Table 3 is respectively organized the comparison of Mus cornea fluorescent staining

3.2Schimer experiment

Result is referring to table 4, as known from Table 4: the comparison of same group different time sections: A group, B group, D organize each time period comparing difference not statistically significant (P > 0.05); C group has statistical significance (P < 0.01) with 1 month comparing difference in 3,5 months; C group comparing difference not statistically significant (P > 0.05) in the time of 3,5 months; E group 3 months and 1 month comparing difference not statistically significant (P > 0.05); E group has statistical significance (P < 0.05) with 1 month comparing difference in 5 months.

The comparison of the different groups of contemporaneity: after 1 month, A, B, C, D, E group difference not statistically significant (P > 0.05); After 3 months, A, B, C, D, E group difference have statistical significance (P < 0.05); B, D group and A group difference not statistically significant (P > 0.05); A, D, E and C group difference have statistical significance (P < 0.05); D and E group difference not statistically significant (P > 0.05); After 5 months, B, D group and A group difference not statistically significant (P > 0.05); D, E group has statistical significance (P < 0.01) with C group difference; D group has statistical significance (P < 0.05) with E group difference.

Table 4 respectively organize Mus Schirmer test experiment reagent paper length measurment comparison ( , mm)

Note: adopt non-ginseng multiple comparisons method (Games-Howell) relatively.

3.3 breakup time of tear film (BUT)

Result is referring to table 5, as known from Table 5: the comparison of same group different time sections: A, B organize 1 month, 3 months, 5 phases of the moons of each time period difference not statistically significant (P > 0.05): C group all has statistical significance (P < 0.01) than difference; D organizes 1,3 month comparing difference not statistically significant (P > 0.05), within 5 months, has statistical significance (P < 0.05) with 1,3 month comparing difference; E group 1,3 and 5 months comparing differences all have statistical significance (P < 0.05).The comparison of the different groups of contemporaneity: A, B group are at the equal not statistically significant of each time period difference (P > 0.05); After 1 month, A, B, C, D, E group difference not statistically significant (P > 0.05); After 3 months, D, E group has statistical significance (P < 0.05) with C group comparing difference; D and E group difference not statistically significant (P > 0.05); After 5 months, C, D, E and A group comparing difference have statistical significance (P < 0.05); D, E and C group difference have statistical significance (P < 0.01); D and E group difference not statistically significant (P > 0.05, table 5).

Table 5 respectively organize the comparison that Mus breakup time of tear film (BUT) measures ( second)

Note: what adopt the multifactor analysis of variance compares (LDS method) relatively between two.

The om observation of 3.4 corneas and lachrymal gland

Normal group (A group): cornea: epithelial layer, anterior elastic membrane, hypothallus, posterior elastic membrane and endodermis are high-visible, epithelial layer is smooth complete; In hypothallus, see a small amount of keratocyte.Lachrymal gland: surface have thin layer connective tissue by every, connective tissue gos deep into essence and glandular tissue is divided into the lobule differing in size.The lymphocyte being dispersed in as seen between lobule and acinus, macrophage and plasma cell.Acinus size evenly, is arranged closely.Sham-operated control group (B group): cornea and lachrymal gland: the substantially same normal group of finding under light microscopic.Surgery models group (C group): cornea: compare C1 group with normal group: epithelial layer cell level increases.Local pinacocyte is shown in irregular a small amount of de-mistake; C3 group: epithelial layer cell level increases, pinacocyte is shown in the de-mistake of irregular lamellar; C5 group: epithelial layer cell level increases, the irregular disappearance of top layer pinacocyte, wing cell exposes; Hypothallus edema thickens, and its inner cell composition increases, and sees massive inflammatory cells infiltrated.Lachrymal gland: compare C1 group with normal group lachrymal gland: acinus endocrine bubble increases to some extent; C3 group: in acinus and acinous cell endocrine increase, see nucleus division phase; C5 group: lachrymal gland acinus is intensive, queueing discipline.In acinous cell, still see a large amount of secretory granule, between acinus, in connective tissue, see that cell component increases. see leaflet core and monocyte infiltration.

Androgen in treating matched group (D group): cornea: compare D1 group with normal group: structure is substantially with normal group lachrymal gland; D3 group: epithelial layer cell level increases, local pinacocyte is shown in irregular a small amount of de-mistake; D5 group: epithelial layer cell level increases.Pinacocyte is shown in the de-mistake of irregular lamellar.Lachrymal gland: compare D1 group with normal group lachrymal gland: substantially with normal group lachrymal gland; D3 group: acinus endocrine bubble increases to some extent: D5 group: in acinus and acinous cell endocrine increase, see nucleus division phase.ZHANGYANMING PIAN treatment group (E group): cornea and lachrymal gland light microscopic and D group are basic identical.

The transmission electron microscope observing of 3.5 lachrymal glands

Normal group (A group) transmission electron microscope observing: visible lachrymal gland cell inner structure is clear, intact nuclear membrane, kernel is obvious, has tight connection between glandular epithelium, the interior rough endoplasmic reticulum of cell is abundant, secreting face has a large amount of secretory vacuoles, and cell surface has microvillus structure; Sham-operated control group (B group) transmission electron microscope observing: finding is substantially with normal group lachrymal gland; Surgery models group (C group) transmission electron microscope observing: C1 group: finding structure is substantially with normal group lachrymal gland; C3 group: nuclear membrane shrinkage, has focal degeneration necrosis (lysosome is black hole shape), mitochondrial swelling; C5 group: acinus queueing discipline, alveolar lumen is seen by each acinus central authorities, the visible a large amount of secretory granule of acinous cell internal upper part. acinous cell basilar part is shown in a large amount of rough endoplasmic reticulums, height and matrix: mitochondrion vacuolation; Visible closely connection between acinous cell; Acinus is asked in connective tissue and is seen a small amount of venule, blood capillary and macrophage, plasmocyte infiltrating, and nuclear membrane shrinkage has focal degeneration necrosis (lysosome is black hole shape).Mitochondrial swelling, intercellular substance broadening. the downright bad shape in cell limit.Focal apoptosis is downright bad, and apoptosis is (karyopycnosis, chromatin margination) obviously.

Androgen in treating matched group (D group) transmission electron microscope observing: D1 group: cavity changes (or having lipoid to drip the change of sample cavity), intercellular substance broadening; D3 group: tube chamber microvillus has and comes off. mitochondrial crista is lost, and has cavity sample to become, and tube chamber has focal lesion, mitochondrial swelling.There is fat to drip sample and become, karyopycnosis, apoptosis is relatively obvious, and blood vessel and endothelium change little; D5 group: the light swelling of mitochondrion, the light swelling of core, focal hydropic degeneration. ambigous nucleus is downright bad. there is inclusion body structure.ZHANGYANMING PIAN treatment group (E group) transmission electron microscope observing: E1 group: mitochondrial swelling, cavity apparition.E3 group: endoplasmic reticulum is relatively abundant, chromatin margination.E5 group: mitochondrial swelling alleviates, loses or becomes cavity.

Four, conclusion

Result shows: ZHANGYANMING PIAN can alleviate lacrimal secretion functional lesion that the male Mus of castration increases the weight of gradually and corneal epithelium and lack and alleviate its local inflammation and react, and xerophthalmia model cornea and lachrymal gland are had to certain protective effect.

By above-mentioned experiment, confirmed that ZHANGYANMING PIAN can obviously increase rabbit xerophthalmia model lacrimal secretion, improve tear film stability and eye table cellularity; The male Mus cornea of xerophthalmia and lacrimal tissue due to castration are had to certain protective effect.Research shows, ZHANGYANMING herbal mixture has significant effect in preventing and treating xerophthalmia disease, and the effect of the xerophthalmia due to its androgen antagonist level declines shows that ZHANGYANMING medicine has the effect of class androgenic.

The above embodiment has only expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.

Claims (3)

1. ZHANGYANMING PIAN is prevented and treated the application in the medicine of xerophthalmia in preparation.

2. application according to claim 1, is characterized in that, described xerophthalmia is video terminal xerophthalmia, mid-aged population xerophthalmia, cataract characteristic of disease xerophthalmia, diabetic xerophthalmia or the unbalance xerophthalmia causing of gonadal hormone.

3. according to the application described in claim 1-2 any one, it is characterized in that, the dosage form of described medicine is tablet, capsule, granule, pill, oral liquid, external preparation or injection.