视网膜色素变性肝肾阴虚证小鼠的模型建立与评价OA

Objective To establish a mouse model of retinitis pigmentosa(RP)with liver and kidney yin deficiency from the perspective of"determining pattern by formula"in Chinese medicine.Methods Forty-eight rd10 mice were used to establish a liver-kidney yin deficiency pattern model through emotional stimulation combined with levothyroxine sodium tablet.The mice were randomly divided into four groups using random number table method:model group(equal volume of normal saline),Leding group(0.15 g/kg of Leding soft capsules),low-dose Ziyin Mingmu Pill(ZYMMP)group(4.5 g/kg),and high-dose ZYMMP group(9 g/kg).Twelve C57BL/6 mice served as the blank group(equivalent volume of normal saline).Each group was given the corresponding drug by intragastric administration once a day for 28 consecutive days.General conditions of the mice were observed,and photographic information of their heads,tongues,claws,and tails were recorded.Serum levels of cyclic adenosine monophosphate(cAMP),cyclic guanosine monophosphate(cGMP)and mouse testosterone(T)were measured using ELISA.The morphological changes in the liver,kidney,and retinal tissues of mice were observed via HE staining,and retinal microvascular morphology was examined using vascular network flattening and periodic acid-Schiff(PAS)staining.Results Compared with the blank group,mice in the model group showed signs of lethargy,dark red and dry tongues with cracks,pale claws and nails,as well as dry and rough tail skin with scaly keratinized patches.Serum cAMP levels increased(P＜0.01),while cGMP and T levels decreased(P＜0.01).Hepatocytes were disorganized with visible atrophy,vacuolar degeneration,and fibrous tissue hyperplasia.Renal tubular cells were irregularly arranged,glomerular capillary walls were defective,and inflammatory cell infiltration and renal interstitial fibrosis were observed.The retina was significantly atrophied and thinner,with loss of retinal pigment epithelial cells and disappearance of the outer nuclear layer structure.Retinal thickness decreased(P＜0.01).Retinal capillaries were disorganized,densely distributed,and thinner in diameter,with noticeable vascular twisting and entanglement.Compared with the model group,Leding group,low-dose and high-dose ZYMMP group showed normal mental state,with red tongue,reddish and moist claws and nails,and a reduction or disappearance of scaly keratinized patches on the tail.cAMP levels significantly decreased(P＜0.01),while cGMP and T levels increased(P＜0.05 or P＜0.01).Liver and kidney cells were more regularly arranged,with reduced fibrous tissue hyperplasia.Retinal thickness significantly increased(P＜0.01).The retinal capillary network was more orderly distributed,with thickened vessel diameters.Compared with the Leding group,the low-dose ZYMMP group showed an increase in cAMP levels(P＜0.01),while both the low-and high-dose ZYMMP groups exhibited a decrease in cGMP levels(P＜0.05,P＜0.01)and an increase in retinal thickness(P＜0.01).Conclusion ZYMMP can improve the general state of mice,reduce serum cAMP levels,elevate cGMP and T levels,and concurrently ameliorate pathological damage in the liver,kidney,and retina,as well as increases retinal thickness.