| Objective:1.Establish diabetic retinal edema model in SD rats.2.The intervention study of Yigan Mingmu Decoction on diabetic retinal edema model in SD rats.3.To explore the protective effect of Müller cells on diabetic retinopathy,which may be realized by regulating autophagy of Müller cells and inhibiting apoptosis.4.To explore whether Yigan Mingmu decoction has protective effect on the organs of the whole body.Methods:1.Part one:to establish diabetic retinal edema model in SD rats.In the blank group,10 SD rats were randomly selected,and the remaining SD rats were used to establish the model.The model was established by intraperitoneal injection of 30 mg·kg-1 STZ.When the blood glucose was stable at 16.7 mmol/L or above,the DM model was established.Three months after modeling,body weight and blood glucose were measured,basic conditions of eyes were observed,Oct,FFA,LC3-Ⅱ and TXNIP were detected,retinal tissue morphology was observed by HE staining,and caspase was detected by IHC The average retinal thickness was higher than 209.78 ± 3.62,the level of he section was not clear,a lot of edema could be seen in each cell layer,and the expression of apoptosis related protein increased,which confirmed the successful establishment of DR and diabetic retinal edema model.2.The second part:To explore the intervention of Yigan MingmuDecoction on diabetic retinoedema model in SD rats.After the establishment of diabetic retinal edema,60 rats were randomly divided into 6 groups with 10 rats in each group:model group,calcium dobesilate group,low-dose Yigan MingmuDecoction group,medium dose Yigan MingmuDecoction group,high-dose Yigan Mingmu Decoction group and autophagy inhibitor group.The blank group and model group were given the same amount of normal saline,and the inhibitor group was given intraperitoneal injection of 3-mA.The body weight,blood glucose,LC3-Ⅱ and TXNIP of SD rats were measured after 4 weeks of gavage.The morphological changes of retina,liver and kidney were observed under light microscope after he staining.The expression of Caspase-3 was detected by IHC.The expression of Caspase-3,TXNIP and LC3-Ⅱ/Ⅰ was detected by Western blot.Results:Part one1.After the successful establishment of diabetic retinal edema model,the weight of the model group was significantly different from that of the blank group,P<0.05,with statistical significance.2.After the successful establishment of diabetic retinal edema model,the blood glucose of the model group was significantly different from that of the blank group,P<0.05,with statistical significance.3.After the successful establishment of diabetic retinal edema model,FFA examination showed microangioma and small bleeding spots in the fundus of the model group,and the fundus structure of the blank group was basically normal.4.After successful modeling of diabetic retinal edema,the contents of LC3-Ⅱ and TXNIP in vitreous cavity fluid of model group were higher than those of blank group(P<0.05).5.The retinal thickness of the model group was higher than that of the blank group(P<0.05).6.After the successful establishment of diabetic retinal edema model,the retinal structure of the model group was disordered after he staining,while the retinal structure of the blank group was clear.7.The expression of Caspase-3 in the model group was higher than that in the blank group(P<0.05).Part two1.After 4 weeks of administration,compared with the blank group,the body weight of rats in model group,positive drug group,low,medium and high dose Yigan Mingmu Decoction group and 3-mA group decreased significantly(P<0.05).2.After 4 weeks of administration,there was no significant change in blood glucose in the blank group.Compared with the blank group,the blood glucose level of the model group increased with the time of gastric Administration(P<0.05),and then remained stable;compared with the model group,the blood glucose levels of the andomin group and the Yigan Mingmu Decoction groups decreased after 4 weeks of gastric Administration(P<0.05);the blood glucose level of the middle dose Yigan Mingmu Decoction group decreased significantly(P<0.05);in addition,the blood glucose level of the 3-mA group decreased It’s not obvious.3.After 4 weeks of administration,the eyegro Ind structure of the blank group was clear,and no microangioma was found;in the model group,some rats could not see clearly,and some rats could see fluorescence leakage and microangioma;in the middle dose Yigan Mingmu Decoction group,the eyeground structure was basically normal compared with other groups except the blank group;in the high-dose Yigan Mingmu Decoction group,3-mA group and low-dose Yigan Mingmu Decoction group,the eyegro und changes were similar,and small microangioma could be seen Hemangioma.4.After 4 weeks of administration,there was no significant change in Oct retinal thickness in the blank group.Compared with the blank group,the OCT retinal thickness in the model group increased significantly with the time of gastric Administration(P<0.05);compared with the model group,the OCT retinal thickness in the andomin group and the Yigan Mingmu Decoction group decreased significantly after 4 weeks of gastric Administration(P<0.05),and the OCT retinal thickness in the middle dose group decreased significantly after 4 weeks of gastric Administration(P<0.05);In addition,there was no significant change in Oct retinal thickness in 3-mA group.5.After 4 weeks of administration,he staining showed that the positive drug group and Yigan Mingmu Decoction groups were improved compared with the model;the middle dose of Yigan Mingmu decoction was significantly improved compared with the former,the boundary was clear,the granular layer was significantly improved,and the retinal edema was significantly reduced;the high dose of Yigan Mingmu decoction was better than the middle dose of Yigan Mingmu decoction The dose was serious and the granular layer was thin.6.After 4 weeks of administration,the expression of Caspase-3 was detected by IHC.Compared with the blank group,the expression level of Caspase-3 in the model group and 3-mA group was significantly increased(P<0.05);compared with the model group,the expression level of Caspase-3 in the positive drug group and Yigan Mingmu Decoction groups was decreased(P<0.05),and it was more obvious in the medium dose group(P<0.05).7.After 4 weeks of administration,compared with the blank group,the expressions of LC3-Ⅱ and TXNIP in the model group were significantly increased(P<0.05);compared with the model group,the expressions of LC3-Ⅱ and TXNIP in the andomin group,middle and high dose Yigan Mingmu Decoction group and 3-mA group were significantly decreased(P<0.05);compared with the low dose Yigan Mingmu Decoction group,the expressions of LC3-Ⅱ and TXNIP in the middle dose Yigan MingmuDecoction group and high dose Yigan Mingmu Decoction group were significantly decreased(P<0.05)Compared with the high-dose Yigan Mingmu Decoction group,the middle dose Yigan Mingmu Decoction group decreased more significantly(P<0.05).8.After 4 weeks of administration,compared with the blank group,the protein expressions of Caspase3,TXNIP and LC3-Ⅱ/lc3-ⅰ in the model group were significantly increased(P<0.05);compared with the model group,the protein expressions of Caspase3,TXNIP and LC3-Ⅱ/lc3-ⅰ in the andomin group and the middle and high dose Yigan Mingmu Decoction group were significantly decreased(P<0.05),and the protein expressions of TXNIP and LC3-Ⅱ/lc3-ⅰ in the 3-mA group were significantly decreased(P<0.05),but caspase3-ⅱ/lc3-ⅰ was significantly increased Compared with low-dose Yigan Mingmu Decoction group,the protein expression levels of Caspase3,TXNIP and LC3-Ⅱ/lc3-ⅰ in middle dose Yigan MingmuDecoction group and high-dose Yigan Mingmu Decoction group were lower,and the protein expression in middle dose Yigan Mingmu Decoction group was lower than that in high-dose Yigan Mingmu Decoction group(P<0.05).9.HE staining of renal tiss ue section showed that the rats in blank group had intact glomerulus,mesangium and basement membrane under he microscope,and there were a lot of glomerular destruction,necrosis and exudative lesions,interstitial hemorrhage in model group;the lesions in andomin group and Yigan Mingmu Decoction group were lighter than those in model group,and interstitial hemorrhage and glomerular destruction were less in medium dose group.Conclusion:1.STZ can induce diabetic retinal edema model in SD rats,which can increase retinal thickness,retinal microangioma,no perfusion area,fluorescence leakage and so on.2.The possible mechanism of Shugan Mingmu Decoction and Huoxue Lishui Yigan Mingmu Decoction in the treatment of diabetic retinal edema is to reduce the relative expression of TXNIP on Müller cells in the retina,make the autophagy level in a balanced state,make the damaged mitochondria produced by oxidative stress through a complete autophagy pathway,avoid the release of inflammatory factors,induce apoptosis and necrosis,and play an important role in the treatment of diabetic retinal edema The protective effect of omentum.3.The Yigan Mingmu decoction has protective effect on important organs. |
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