Clinical And Mechanism Research About The Treatment Of Xeroma Used Runmuling

BackgroundXeroma is a syndrome characterized by absolute or relative deficiency of tear film aqueous constituents, abnormal tear distribution on the ocular surface and increased tear evaporation, which are caused by such factors as reduced aqueous tear production, inappropriate mucus secretion, meibomian gland dysfunction(MGD),etc. Xeroma patients often complain of discomforts such as sensation of dryness and foreign body in the eye, photophobia, blurred or fluctuating vision, and so on. Some serious ones may suffer from an obvious decline of eyesight that affect their work and life. For the time being the cause and mechanism of xeroma are not fully known to us. The methods of testing are to be improved, and effective treatments are still not present. The application of immunosuppressive agent and incretion, the innovation of surgery and the introduction of testing new instruments all bring hope to the treatment of xeroma. Western medicine mainly adopts a symptomatic and local treatment, which uses artificial tear substituent and eternal or temporary blockage of tear duct. to preserve tear so that clinical symptoms can be alleviated, but an ultimate curative effect cannot be achieved. The clinical application of Cyclosporine A is limited because of its apparently stimulant effect. The autologous submandibular gland transplantation is a surgery trauma anyway. We have been exploring new effective methods to treat xeroma, which not only are atraumatic or causing as little trauma as possible, but also stimulate the tear glands to secrete more tear. Therefore, it is an urgent task to find a new treatment of xeroma that is reliable and has a satisfactory effect. Traditional Chinese medicine is the answer that meets the above requirements. We have found in the literature of traditional Chinese medicine that Bidens Bipinnata L. has "a side effect of inducing excessive tear", which inspired us to develop Runmuling, an oral granule preparation composed of Bidens Bipinnata L., medlar and chrysanthemum. The remedium cardinale in the prescription, Bidens Bipinnata L., can clear away heat and toxic material, detumescence and eliminate stasis to activate blood circulation; chrysanthemum is for lung and liver channel tropism, which can remove heat, eliminate toxins and nourish the liver to improve eyesight and stop tears; medlar is for liver and kidney channel tropism, which can invigorate the liver and kidney and nourish the Yin to improve eyesight. The mixture of the three of them can have a combined effect of treating and curing xeroma. Clinical experiments have proved its quite good curative effect. To further our understanding of the prescription, the following clinical and animal experiments have been conducted to verify its curative effect, and to explore its mechanism.Part One Clinical research Clinical Observation of the Effect of Tradition Chinese Medicine Runmuling on XeromaResearch Purposes:To observe the extent to which the symptoms and signs of xeroma changes prior to and after Runmuling treatment, and compare the Runmuling group and the placebo group in order to investigate the exact curative effect and security of Runmuling on xeroma.Methods:75 patients with xeroma are investigated randomly and divided into parallel groups for comparison. They are randomly divided into Runmuling group (37 cases), and Placebo group (38 cases). The Runmuling group takes a packet of instant granules made of Bidens Bipinnata L. (15g), medlar (10g) and chrysanthemum (6g) each time and twice a day. The placebo is produced by the same manufacturer in a double-blind and double simulation method and taken by the placebo group with exactly the same dosage as Runmuling group. The treatment lasts for 8 weeks. Since the beginning of the treatment, every other week the patients are observed and evaluated in terms of symptom score, SIT, BUT, corneal staining and adverse reaction.Results:1.Prior to the treatment there is no statistically significant difference between the two groups in gender, age, tear flow, breakup time of tear film, corneal fluorescein staining and symptom score (P>0.05);nor in eye dryness, redness and fatigue (P>0.05); However, there is some statistically significant difference between the two groups in foreign body sensation and photophobia, with the Runmuling group more serious than the placebo group, signaling that the Runmuling group has more highly graded patients than the placebo group. The main factors which affect curative effects are balanced and comparable for the two groups.2. After 8 weeks of treatment, the Runmuling group has achieved 62.2% highly effective rate,24.3% effective rate and 13.5% ineffective rate; the placebo group has 28.9% highly effective rate,50% effective rate and 21.3% ineffective rate. Both the highly effective rate and the total effective rate of the Runmuling group are higher than those of the placebo group, and the difference between the two groups are statistically significant (P<0.05).3. After the Runmuling treatment, the SIT increases, the BUT prolongs, the eye dryness symptom is alleviated, and the difference prior to and after the treatment is statistically significant (p<0.01).4. SIT:After 8 weeks of treatment, the mean SIT value of both groups increases, and there is no statistically significant difference between the two groups'mean SIT increase (the Runmuling group being 1.47±3.86mm/5min, and the placebo group being 0.93±5.34mm/5min). BUT:after 8 weeks of treatment, the increase of the mean BUT value of the Runmuling group (1.05±1.86s) is larger than that of the placebo group, and the difference is statistically significant (P<0.01).5. eye symptoms:after 8 weeks of treatment, the Runmuling group does better in alleviating eye dryness and fatigue, and the difference is statistically significant (P<0.05). Specifically, there is statistically significant difference between the two groups in the symptom of eye dryness after 6 weeks of treatment, with the Runmuling group superior to the placebo group.6. During the course of the research neither local nor systemic adverse reaction has been found, and none of the patients suffers from eyesight decline.Conclusion:1.Runmuling can promote SIT, lengthen BUT, and alleviate eye dryness, and has an obvious therapeutical effect on xeroma.2. It is safe to use Runmuling for xeroma treatment.Part Two Experimental research Study on the Effect of the Water-extract Solution of Bidens Bipinnata L. on muscarinic-receptorsResearch Purposes:To investigate the effect of water-extract solution of Bidens Bipinnata L. on muscarinic-receptors in the guinea Pig ileum.Methods:The cumulative dose-response curve method is adopted to record the contraction of isolated longitudinal ileal muscles of guinea pigs, using the mean contractile force (MCF) of muscarinic-receptors as the statistical indicator to compute the percentage variation of ileum contraction prior to and after the treatment.1. Adding 100μl water-extract solution of Bidens Bipinnata L. each time to make solutions with different final concentrations, including 10,100,1000μg/ml respectively. For the control group, physiological saline is added. The ileum contractions of both groups are recorded.2. Adding atropine solution with a concentration 10-8mmol/L into the baths of both groups, and 10 minutes later adding 100uml water-extract solution of Bidens Bipinnata L.to make solutions with final concentrations of 10,100,1000μg/ml. For the control group, physiological saline is added. The ileum contractions of both groups are recorded.3. Using the cumulative dose method to add 100μl methylene dichloride fraction of the water-extract solution of Bidens Bipinnata L. each time, at an interval of 5 minutes, to make solutions with concentrations of 0.0625g crude drug/L,0.125g crude drug/L,0.25g crude drug/L,0.5g crude drug/L,1g crude drug/L. The cumulative dose-response curve is drawn. Tyrode's solution is used to rinse the specimen until the contractile force returns to normal. The same process is applied to ethyl acetate fraction, normal butanol fraction, water fraction, concentrated water-extract solution of Bidens Bipinnata L. and dimethyl sulfoxide (DMSO) respectively, and equivalent curves are then drawn.Results:1. water-extract solution of Bidens Bipinnata L. can significantly promote the contraction of isolated ileal muscles of guinea pigs; the conctractile force is in proportion to the concentration, with MCF increased by 32.7% at a concentration of 1000ug/ml. the difference between the experiment group and the control group is statistically significant (P<0.001).2. The contraction suppressive percentage of atropine control group and atropine/water-extract solution of Bidens Bipinnata L. group is 27.4% and 10.1% respectively. After water-extract solution of Bidens Bipinnata L. treatment, the suppressive effect of atropine on the contraction of isolated ileal muscles is reduced, and the difference is statistically significant.3. Water-extract solution of Bidens Bipinnata L. can significantly excite the contraction of isolated longitudinal ileal muscles of guinea pigs. It strengthens the ileul contraction and increase the MCF. The dose-response relation is significant. The active ingredients of Bidens Bipinnata L.'s in exciting ileul muscles are concentrated in its water fraction and normal butanol fraction.Conclusions:Water-extract solution of Bidens Bipinnata L. can significantly reduce the suppressive effect of atropine on the contraction of isolated longitudinal ileal muscles of guinea pigs, which shows that water-extract solution of Bidens Bipinnata L. has an antagonistic effect on atropine and an exciting effect on muscarinic-receptors.