Exploring The Mechanism Of Biological Rhythm Disorder And The Intervention Effect Of Wumei Pill In POAG Based On Intraocular Pressure Curve

Part Ⅰ Visual knowledge mapping of glaucoma and biological rhythmObjectiveThe visual knowledge mapping technology was applied to systematically integrate and analyze the knowledge included in the correlation study of glaucoma and biological rhythm,so as to clarify the correlation between glaucoma and biological rhythm,and to mine the targets of TCM intervention in biological rhythm disorder.MethodsFirstly,the clinical or animal experimental studies related to glaucoma and biological rhythm since the establishment of CNKI/WANFANG/VIP/PubMed/WOS core collection were systematically reviewed;secondly,the knowledge was extracted through literature analysis and the knowledge model of "glaucoma biological rhythm" was constructed with TCMKOS;finally,Neo4j was used for visual knowledge map analysis.ResultsA total of 48 articles were included as knowledge sources through literature studies,including 33 clinical studies and 15 animal experiments.14 kinds of entities with diseases,intervention measures and observation indicators as the core were established,and 98 examples were created,including 18 examples of diseases,43 examples of intervention measures and 37 examples of observation indicators.Finally,the "glaucoma-biological rhythm" knowledge model was constructed and visually analyzed.ConclusionsGlaucoma may destroy the regulation system of "intrinsic photosensitive retinal ganglion cell-suprachiasmatic nucleus-neuroendocrine",leading to biological rhythm disturbances mainly manifested by intraocular pressure rhythm disorders,sleep disorders,and mood disorders.Part Ⅱ The correlation between the characteristics of 24h IOP rhythm and"Jueyin disease ready to release" in patients with POAGObjectiveThe characteristics of intraocular rhythm in patients with primary open angle glaucoma(POAG)and the correlation revealed the correlation between glaucoma and visual field defect.Methods24-hour intraocular pressure monitoring in 43(86 eyes)with ocular hypertension primary open-angle glaucoma(POAG),The measurement time points are 2:00,5:00,7:00,10:00,14:00,18:00,22:00.Based on the obtained 24h IOP,24h IOP fluctuation parameters(mean,peak,trough,and difference),"Jueyin disease ready to release" IOP fluctuation parameters(mean,peak,trough,and difference in 2:00,5:00,7:00)and out of "Jueyin disease ready to release" IOP fluctuation parameters(mean,peak,trough,and difference in 10:00,14:00,18:00,22:00).Visual Field defects were classified as mild(VFI>91%),moderate(91%≥VFI>78%)and severe(VFI<78%)based on Visual Field Index(VFI)according to USP-GVFSS Visual Field grading method.Results1.The characteristics of the fluctuation curve of intraocular pressure(IOP)in POAG patients:(1)the IOP of mild patients increased in the morning,the peak of IOP appeared at 7:00(11 eyes,46%),then decreased gradually,a small peak appeared at 2:00 pm,and the trough value appeared at 18:00 pm;(2)IOP fluctuated slightly in moderate patients,the peak time was 5:00(5 eyes,22%),7:00(5 eyes,22%),the trough value appeared at 10:00 am;(3)IOP increased significantly in severe glaucoma patients at 2:00(13 eyes,33%),at 5:00(8 eyes,21%)and 7:00(6 eyes,16%),the IOP decreased gradually after 7:00.The peak time was consistent with that of the mild patients,and appeared at 18:00.2.The characteristics of 24-hour iop rhythm in different degrees:(1)Mean:severe>mild and moderate(severe vs.moderate,P<0.001;severe vs.mild,P=0.001);(2)Peak:severe values>mild and moderate(severe vs.moderate,P=0.002;severe vs.mild,P=0.003);(3)Valley:severe>mild and moderate(severe versus moderate,P=0.013;severe versus mild,P=0.027);(4)Difference:there was no significant difference in diurnal IOP difference between mild,moderate and severe(P>0.05)3.The characteristics of IOP rhythm at "Jueyin disease ready to release":(1)Mean:Severe>moderate,mild(severe vs moderate,P=0.035;severe vs mild,P<0.001);(2)Peak:severe>mild,moderate(severe vs.moderate,P=0.011;severe vs.mild,p<0.001);(3)Valley:severe valley>mild valley(P=0.006);(4)Difference:for nocturnal difference of iop,severe>mild(P=0.006),there was no significant difference between moderate,mild and severe IOP(P>0.05),there was no significant difference between moderate,mild and severe degree(P>0.05).4.The characteristics of IOP rhythm out of "Jueyin disease ready to release":(1)Mean:severe>moderate(P<0.001),there was no significant difference between mild and moderate,severe(P>0.05)(2)Peak value:moderate<severe,mild(severe vs.moderate,P=0.003;mild vs.moderate,P=0.009),there was no significant difference between mild and severe(P>0.05);(3)Valley:severe>moderate,there was no significant difference between mild and moderate,severe(P>0.05),(4)Difference:there was no significant difference between mild,moderate and severe(P>0.05).5.The correlation between IOP rhythm and visual field index(VFI):(1)there was a linear regression relationship between IOP at 10:00 IOP,2:00 IOP and VFI;(2)there was a weak negative correlation between IOP Difference and VFI at"Jueyin disease ready to release".ConclusionsThe 24-hour IOP rhythm disorder in POAG patients was obvious from "Jueyin disease ready to release".The disturbance of IOP rhythm is more significant in severe glaucoma patients.Based on the theory of "rhythm and stability",this part of the study confirms the scientific nature of "rhythm disharmony" in glaucoma from the perspective of intraocular pressure rhythm.Part Ⅲ Intervention effect and mechanism of Wumei pill in chronic hypertension ratsObjectiveTo evaluate the intervention effect of black Wumei pill on apoptosis and biological rhythm disorder of RGCs in chronic hypertension rats(COH)model,reveal the mechanism of multiple targets to regulate biological rhythm and protect visual function,and clarify the scientific connotation of" Jueyin disease ready to release".MethodsRetinal morphological changes in COH rats were observed by HE staining,the apoptosis rate of RGCs was observed by TUNEL staining,2:00,8:00,14:00,20:00,ELISA by 2:00 MT/CORT/ACTH/CRH secretion,the expression of ipRGCspecific protein melanopsin was detected by WB,The expression of the retinal core clock gene Clock/Bmal 1/Per 1/Cry 1/Rev-erba mRNA was determined by RTPCR.Results1.Wumei pill can improve retinal morphological changes and inhibit apoptosis of RGCs in COH rats(1)Retinal histomorphological changes:28-day COH rats damaged retinal structure,loose inner and outer nuclear layers,RGCs disorder,the number,RNFL became thin and stained,and the nipple was obviously depressed;the Wumei pill group was clear and complete,the RGCs were lost,the RNFL was slightly thinner than the normal group,and the optic disc was slightly depressed.(2)Apoptosis of RGCs:the apoptosis rate in COH model group was significantly higher than that in blank group,statistically significant(P<0.001);the apoptosis rate in Wumei group was lower than the model group,and the difference was statistically significant(P<0.001).2.Wumei pill may improve the biological rhythm disturbance in COH rats(1)The rats reduced IOP and IMP by 2:00:compared with the model group,2:00 IOP(P<0.001);the IOP difference,P<0.001 compared with the blank group and P=0.028 compared with the model group,all the differences were statistically significant.(2)2:00 plasma MT/CORT/ACTH/CRH secretion level:ELISA:2:00 plasma MT/CORT/ACTH/CRH secretion level showed that the 2:00 MT secretion level compared with the control group,statistically significant(P<0.05);2:00 MT/CRH/CORT expression in COH rats,which increased MT and CORT expression and reduced CRH expression level,statistically significant(P<0.05).3.Mechanism of action of Wumei pill on biological rhythm disorder in COH rat model(1)ipRGCs-specific marker protein--melanopsin expression:detected by Westernblot,the results showed that the relative expression level of melanopsin in the model group decreased significantly compared with the blank control group,and the difference was statistically significant(P<0.001);compared with the model group,the relative expression level of melanopsin in the Wumei group increased,and the difference was statistically significant(P=0.003).(2)Expression of retinal core clock gene:the relative expression of Clock/Bmal 1/Cry 1/Per 1/Rev-erba mRNA was detected by qRT-PCR,The results show that,when compared to the blank group,Clock/Bmal1/Per 1/Cry 1/Rev-erbα mRNA expression was decreased in the model group,The difference was statistically significant(P<0.05);Compared to the model group,The relative expression level of Clock/Bmal 1/Cry/mRNA increased significantly in the Wumei pill group,Statistically significant difference(P<0.05 or P<0.001).Conclusions1.Burning method on the sclera vein induced chronic high intraocular pressure rat model can lead to 28 days ipRGCs melanopsin specificity protein expression is reduced,the retinal Clock/Bmal1 Per1 fires our/Cry1/Rev-erba mRNA expression level decreased relatively,abnormal plasma MT secretion by 2:00 PM Sunday,and intraocular pressure increase volatility,which could induce RGCs apoptosis.It is proved that "rhythm and stability"in glaucoma is scientific at the molecular level.2.Based on the theory of"rhythm and stability",jue Yin disease main square Wumei pill can be mediated melanopsin protein expression and regulation of the core Clock/Clock gene Bmal1/Cry1,biological rhythm disorder,thus improving COH model rats RGCs apoptosis inhibition.