¡¡¡¡RESULTS

¡¡¡¡Basic Tear Secretion The comparison of S¢ñt measured value in normal group (A1, A2, A3): according to the variance analysis , there was no significant difference among group A1, A2 and A3 (Table 1, P>0.05), and there was also no obvious difference for rats¡¯ S¢ñt of its preª² and postª² self ª²match t test , before and after the use of physiologic brine eye drops. (P>0.05). The comparison of S¢ñt measured value in model group (B1, B2, B3): the model group was castrated respectively for 1, 2, 3 months both proª² and postª²self ª²match t test, the basic tears secretion volume had decreased obviously and the wetness length of filter paper had shortened .The changes were significant (P<0.01,P<0.01,P<0.01) .Moreover, by variance analysis, after being castrated, the tears secretion volumes of groups B2,B3 were even lower (P<0.05,P<0.01), compared with that of group B1.The comparison revealed that the basic tears secretion volume was on the decline with the increase of time.

¡¡¡¡The comparison of S¢ñt measured value in therapy group (C1, C2, C3): the castrated rats were treated with extract of Buddleja officinalis eye drops. The results were by variance analysis and indicated that there was no obvious difference between C1, C2 and C3(P>0.05). Moreover, by variance analysis, the S¢ñt values of group C had apparent difference (P>0.05, P>0.05, P>0.05) ,which showed the eye drop significantly kept basic tears secretion volume.

¡¡¡¡The contrast of the S¢ñt value between group C and group B(C1 vs B1, C2 vs B2, C3 vs B3): Contrasted with the former wetness length of filter paper, the basic tears secretion volume of group C had greatly increased through the therapy of eye drops. By the variance analysis , the wetness length of filter paper of group C was apparently longer than that of group B(P<0.01, P<0.01, P<0.01).Table 1 S¢ñt values before and after the use of eye drops(ÂÔ)Table 2 BUT values before and after the use of eye drops(ÂÔ)

¡¡¡¡Tear Film Stability The contrast of BUT values of sham group(A1,A2,A3):by variance analysis, there were no apparent differences among A1,A2 and A3 (P>0.05). Furthermore, by t investigation, the BUT values of various groups showed no obvious changes after the use of natural saline (P>0.05). The contrast of BUT values of model group(B1£¬B2 and B3):by t investigation, the tear film stability of model group was on the decrease after being castrated respectively 1, 2 and 3 months later. The BUT value was longer than before(P<0.01, Table 2). Moreover, compared with the tear film stability of group B1,the BUT value of group B3 was apparently even lower (P<0.05)after being castrated, showing that the tear film stability was on the decline with the time. The comparison of BUT values of therapy group(C1,C2 and C3):by variance analysis, there were no obvious differences in BUT values among groups C1,C2 and C3 after the treatment of eye drops(P>0.05). And¡¡by t investigation, the BUT values of various groups showed no difference after the use of eye drops(P>0.05, P>0.05, P>0.05), indicating that the extract of Buddleja officinalis eye drops significantly kept tear film stability. The comparison of BUT values between therapy group and model group(C1 vs B1, C2 vs B2,C3 vs B3): compared with the former BUT, the tear film stability of groups C1,C2 and C3 had increased after the treatment of eye drops by variance analysis.The BUT had increased apparently contrasted with the corresponding model groups B1,B2 and B3(P<0.01,P<0.01,P<0.01).Furthermore,by variance analysis, compared with the tear film stability of group C1 castrated 1 month later, the BUT of group C3 castrated 3 months later was relatively lower(P<0.05).The results proved that the eye drops¬ð capability for keeping tear film stability was on the decrease with the time.Table 3 Positive expression of AR values(ÂÔ)

¡¡¡¡Lacrimal Gland¬ðs Androgen Acceptor Investigated by FCM, the AR positive expression values of various groups were listed in Table 3. The contrast of AR positive expression values of sham group (A1, A2 and A3): By variance analysis, there were no obvious differences among groups A1, A2 and A3.

¡¡¡¡The contrast of AR positive expression values of model group(B1,B2 and B3):By variance analysis, compared with the AR positive expression values of groups B1 and B3, the value of group B2 castrated 2 months later is relatively higher(P<0.05, P<0.05).The results indicated that the AR positive expression values had the tendency of decreasing, then rising followed by decreasing again with the time. The contrast of AR positive expression values of therapy group(C1,C2 and C3):By variance analysis, compared with the AR positive expression value of group C1 castrated 1 month later,the value of group C3 castrated 3 months later was relatively lower(P<0.05).Maybe the tissue adapt gradually to the androgenª²like effects of the flavonoids of Buddleja officinalis and the easing feedback adjustment caused the results. The contrast of the AR positive expression values between therapy group and model group(C1 vs B1,C2 vs B2 and C3 vs B3):By variance analysis,the AR positive expression values of groups C1,C2 and C3,receiving the treatment of Buddleja extract eyedrops after being castrated,increased apparently compared with the corresponding model group B1,B2 and B3(P<0.01, P<0.01, P<0.01).The results indicated that the flavonoids of Buddleja had pretty good androgenª²like effects.

¡¡¡¡DISCUSSION

¡¡¡¡Chinese medicine is an ideal source for treating dry eye for Chinese medicine provide a large variety of natural herbs to choose from as well as the high safety of herbal hormones in clinic use. However, the research is new from the perspective of Chinese medicine to treat dry eye; the research is lack of depth and extent, no drugs for special treating dry eye, no further modern study dry eye which combines theories and practices of Chinese medicine, no safety measurements and evaluations for dosage of eye drops and no consideration for the fake signs of worsening dry eye caused by the eye drops. It has been shown in the previous research that the flavonoids in Buddleja officinalis have sound experimental effects to the animal models to treat dry eye caused by castration, which may relate to the androgenª²like effects of the flavonoids in Buddleja officinalis, but further researches are needed to make clear whether the mechanism is similar and how it acts[2ª²5]. This experiment aims at proving the androgenª²like effects of eye drops made from extract of Buddleja officinalis in lacrimal gland and the curative effects of dry eye caused by the declining of androgen level via the research of androgenª²like effects of eye drops made from extract of Buddleja officinalis in lacrimal gland.

¡¡¡¡Dry eye hasn¬ðt got a definitive name in TCM. According to the degree of syndromes it can be classified into slight level ¡°baisezheng¡±, and the heavy level ¡°Less Aqueous Humor¡± and ¡°waizhangyizheng¡±. The dry eye caused by gonadal hormones level imbalance should be classified into ¡°Less Aqueous Humor¡±, ¡°waizhangyizheng¡± is the result of deterioª²ration. Mizhuan Yanke Longmulun states "What is dry eye? It is caused by the heat of zang and fu organs, the waterway is dry, the heat transfer to the defense, the genuine qi cannot nourish the eye. This is the reason." Therefore, "Less Aqueous Humor" is caused by damage of yin by the inner heat, and the heat transfer into Windª²heat externally. Suwen xuanmingwuqipian says "five zang organs transfer into fluid, and liver into tear." Yinhaijingwei also says "tear is the fluid of liver". From these we can see that "shenshuijiangku" is rooted in liver, so the principle for treatment should focus on clearing the liver heat, nourishing the liver yin and relieving the Windª²heat externally.

¡¡¡¡Buddleja offcinalis can clear and nourish the liver, it is the right drug for liver meridian of jueyin[6]. Yinhaijingwei Yaoxinlun says "Buddleja offcinalis acts on liver meridians and brightens the eyes" The representative dosage collected is Mimenghuasan with Buddleja offcinalis as its main herb.Yinhaijingwei Juanzhixia says "Mimenghuasan: for treatment the lingering internal and external zhangyi, fear of the sunshine qi stagnation caused by Windheat." Mizhuan Yanke Longmulun has some statement for treat dry eye: "Mimengª²huasan: for treat the attack of wind, dizziness and fear of the sunshine. The eye is hard to open, itching or aching dizziness, dry, dull pain with red swelling and aching eye can be treated by it." From this we can see that Buddleja offcinalis can clear the liver heat and supplement liver yin internally, and relieve the Windª²heat of the defense externally. This is the mechanism of dry eye in TCM. With the theory of TCM, it is an ideal drug for dry eye.

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