The Effect And Mechanism Of Yiqi Bushen Recipe On Retinopathy Of Diabetic Rats

Part one: Systematic evaluation of the treatment of diabetic retinopathy by invigorating qi and tonifying kidneyObjective: To systematically evaluate the clinical effect of supplementing qi and kidney in the treatment of diabetic retinopathy by Meta analysis.Methods: The foreign literatures were retrieved from PubMed,Cochrane Library and Embase,and the Chinese literatures were retrieved from CNKI,VIP and Wanfang medical databases.All literatures of randomized controlled clinical trials(RCTS)on the treatment of diabetic retinopathy by supplementing qi and kidney were collected from the self-built database until January 30,2022.Two reviewers including the author independently analyzed and included the articles,and Revman5.4 software was used for systematic evaluation.Results: A total of 13 RCT studies were included,including 1227 patients.Clinical efficacy,visual acuity,fundus examination score,TCM syndrome score,macular fovea thickness,vascular endothelial growth factor(VEGF),tumor necrosis factor-α(TNF-α),and incidence of adverse events were included in the meta-analysis.1.Compared with the control group,the clinical efficacy of invigorating qi and toning kidney was significantly improved: [OR=3.60,95%CI(2.72,4.77),Z=8.93,P<0.00001];2.The qi-tonifying and kidney-tonifying group was more effective in improving visual acuity than the control group:[MD=0.19,95%CI(0.10,0.28),P<0.00001];3.Compared with the control group,the fundus examination score in the invigorating qi and tonifying kidney group was effectively reduced: [MD=-0.31,95%CI(-0.37,-0.25),P<0.00001];4.Compared with the control group,the qi-tonifying and kidney-tonifying method could effectively reduce the TCM syndrome score: [MD=-5.42,95%CI(-7.98,-2.85),P<0.00001];5.Compared with the control group,the thickness of macular fovea in invigorating qi and toning kidney group was effectively reduced: [MD=-20.18,95%CI(-30.79,-13.57),P<0.00001];6.Compared with the control group,the method of invigorating qi and tonifying kidney could effectively reduce VEGF level: [MD=-34.84,95%CI(-41.12,-28.56),P<0.00001];7.Compared with the control group,the level of TNF-α in invigorating qi and tonifying kidney group was effectively reduced: [MD=-16.46,95%CI(-26.15,-6.76),P<0.001];8.Compared with the control group,the method of invigorating qi and tonifying kidney could effectively reduce the occurrence of adverse events: [OR=0.45,95%CI(0.21,0.95,Z=2.10,P<0.05).Sensitivity analysis shows that the results of this study are robust and reliable.Conclusion: The treatment of diabetic retinopathy with qi-tonifying and kidney-tonifying method has a significant therapeutic effect,its clinical efficacy and visual acuity are better than the control group,and can reduce fundus examination score,TCM syndrome score,macular fovea thickness,VEGF level,TNF-α level,and reduce the incidence of adverse events,which has a high clinical value.Part two: Effect and mechanism of Invigorating Qi and tonifying Kidney prescription on diabetic rat retinopathyObjective: To study the effect of Yiqi tonifying kidney formula on diabetic retinopathy in rats and explore its possible mechanism.Methods: sixty healthy male SD rats were randomly divided into control group(group C),model group(M),Yiqi Bushen prescription low-dose group(group L)and Yiqi Bushen prescription high-dose group(group H),with 15 rats in each group.After 1 week of adaptive feeding,except for the control group,the rats in the other three groups were intraperitoneally injected with streptozotocin solution diluted with sodium citrate solution 60 mg for modeling.Group L and GROUP H were intragastrated with 200mg/kg/d and 800mg/kg/d suspension of Yiqi and Kidney Tonifying formula,respectively.Group C and GROUP M were intragastrated with equal volume ultra-pure water.The duration of intragastric administration was 8 weeks,during which the general state,body weight and blood glucose of the rats were recorded weekly.After the experiment,the serum,eyeball and retina tissues of rats were taken for study.Total cholesterol(TC),triglyceride(TG),serum cystatin C(CYS-C),uric acid(UA)and basic indexes of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea nitrogen(BUN)and creatinine(Cr)related to diabetic retinopathy were detected in serum.Retinal pathological sections were taken to observe the changes of retinal pathological tissue.Qrt-pcr was used to detect the mRNA expression of monocyte chemotactic protein-1(Mc P-1),TNF-α,heme oxygenase 1(HO-1),nuclear factor E2-related factor 2(Nrf2),VEGF and glial fibrillary acidic protein(GFAP)in retinal tissue.Western Blot was used to detect the expression of GFAP and VEGF in retinal tissues.Results: 1.General condition: The rats in group C were generally in good condition with free movement and transparent lens;In group M,the hair color was dull,the activity was poor,the lens was cloudy,and the volume of feces was large.Compared with M group,the general state of rats in L and H groups was improved to a certain extent.2.Weight of rats: With the increase of weekly age of intragastric administration,weight of rats in group C increased gradually;The weight of rats in M group,L group and H group fluctuated significantly,showing emaciation,and there was no statistical difference in body weight among the three groups(P>0.05).3.Blood glucose of rats: Blood glucose of rats in group C was normal and stable;The blood glucose levels in M,L and H groups were significantly higher than those in C group(P<0.001).Compared with group M,blood glucose in group L showed a downward trend,but only showed statistical difference at week 1(P<0.05),while blood glucose in group H decreased significantly from week 4(P<0.05).There was no significant difference in blood glucose between GROUP L and group H(P>0.05).4.Serum biochemical indices: At 8 weeks,compared with group C,the concentrations of TG,TC,Cys-C and UA in group M were significantly up-regulated(P<0.05);Compared with group M,the concentrations of TC and UA in group L were decreased(P<0.05),while the concentrations of TG,TC and UA in group H were decreased,with statistical significance(P<0.05).There were no significant differences in the concentrations of TG,TC,Cys-C and UA between GROUP L and group H(P>0.05).5.Liver and kidney function: there were no significant differences in serum ALT,AST,BUN and Cr concentrations among all groups(P>0.05).6.Retinal tissue staining: the inner boundary membrane of the retinal tissue of the rats in group C was intact,no capillary endothelial cells were embedded,single layer of ganglion cell layer was closely arranged,no ganglion cell loss was observed,and the other layers were neatly arranged with clear structure.The inner boundary membrane of the retina of rats in group M was swollen and uneven,capillary endothelial cells were embedded,ganglion cell layers were disordered,ganglion cells were lost or migrated to the inner boundary membrane,and cells in the inner and outer nuclear layers were disordered and the structure was loose.Compared with group H,the swelling of inner boundary membrane was significantly reduced in group L and group M,and no capillary endothelial cells were embedded.The ganglion cell layers were arranged relatively regularly,and the loss of ganglion cells was reduced,while the cells in other layers were arranged more orderly.7.Effects of Yiqi Bushen Prescription on the mRNA expressions of Mc P-1 and TNF-Α in retina: compared with group C,the mRNA expressions of Mc P-1 and TNF-Α in retina of group M were significantly increased(P<0.05);Compared with M group,the mRNA expressions of Mc P-1 and TNF-Α in retina of L and H groups were decreased(P<0.05);Compared with L group,the mRNA expressions of Mc P-1 and TNF-Α in retina of GROUP H were significantly decreased(P<0.05).8.Effects of Yiqi Bushen Prescription on the expression of RETINAL HO-1 and Nrf2 mRNA: Compared with group C,the expression of retinal HO-1 and Nrf2 mRNA in group M was increased(P<0.05);Compared with M group,there was no significant change in L group(P>0.05),but the mRNA expressions of HO-1 and Nrf2 in H group were significantly up-regulated(P<0.05).Compared with L group,the mRNA expressions of RETINAL HO-1 and Nrf2 in GROUP H were significantly up-regulated(P<0.05).9.Effects of Yiqi Bushen Prescription on the mRNA expressions of GFAP and VEGF in retina: compared with group C,the mRNA expressions of GFAP and VEGF in retina of group M were significantly increased(P<0.05);Compared with M group,GFAP mRNA expression in L group was significantly down-regulated(P<0.05),while VEGF mRNA expression was not significantly different(P>0.05),while GFAP and VEGF mRNA expression were significantly down-regulated in H group(P<0.05).Compared with L group,GFAP and VEGF mRNA in H group were significantly down-regulated(P<0.05).10.Effect of Yiqi Bushen Prescription on the expression of GFAP and VEGF protein in retina of rats: compared with group C,the expression of GFAP and VEGF protein in retina of group M was up-regulated(P<0.05);Compared with M group,GFAP protein expression in L group showed a downward trend,but there was no statistical significance(P>0.05),while VEGF protein expression was significantly down-regulated(P<0.05).The expressions of GFAP and VEGF protein in retina of GROUP H were significantly down-regulated(P<0.05).Compared with L group,the protein expressions of GFAP and VEGF in retina of group H were down-regulated(P<0.05).Conclusion: 1.The prescription for supplementing qi and tonifying kidney can improve the general condition of diabetic rats,reduce the levels of blood glucose,serum total cholesterol,triglyceride and uric acid,and improve the retinal histopathological damage caused by diabetes.2.Yiqi tonifying kidney formula can reduce the expression of retinal inflammatory factors(Mc P-1,TNF-α)and up-regulate antioxidant related factors(HO-1,Nrf2)in diabetic rats,thereby reducing the expression of retinal vascular endothelial growth factor and glial fibrillary acidic protein,and protecting retinal microvessels and neurons.