| Background:Cassiae Semen is a traditional Chinese medicine,which has long been used as the treatment of diabetic hyperlipidemia and diabetic constipation in clinic.Naphtopyrones and anthraquinones in Cassiae Semen might be responsible for the treatment of diabetes and its complications.Objective: The present study was designed to investigate the optimization of extraction procedure,the quality control and the anti-diabetic effects of Cassiae Semen extract(CSE).Methods: The optimization of preparation procedure of CSE was performed using single-factor and orthogonal tests with aurantio-obtusin as the indicator.Morphological identification of color and odor of CSE was observed.The content of cassiaside,rubrofusarin-6-O-β-D-gentiobioside,glucoaurantio-obtusin,cassiaside C and aurantio-obtusin in CSE was determined using HPLC.CSE were orally administered at 27,54 and 81 mg/(kg·d)dose to high-sucrose-high-fat diet and streptozotocin(STZ)-induced diabetic rats for 60 days.Body weight,glucose metabolism profiles,lipid metabolism profiles and histopathological examination of pancreas were measured to assess the anti-diabetic effect of CSE.Oxidative stress markers and inflammatory factors were measured using commercial kits.Renal function related parameters were also measured.Histopathological examination of kidney was conducted for the validation of pathological changes in diabetic rats.Immunohistochemical examination of kidney was measured to investigate the expression of RAGE in renal tissues.Histopathological examination of liver and retina was conducted for investigation of protective effects of CSE in liver and retina in diabetic rats.Results: CSE was obtained with a yield of about 4% using D101 macroporous resin purification after 80% ethanol extraction.CSE was a brown-black powder with a slight bean odor and a slight bitter taste.Five compounds,including two anthraquinones and three naphtopyrones were determined,and their contents accounted for about 20% in CSE.The content limit was also established.After administration for 60 days,compared with diabetic control,groups treated with CSE exhibited an increased body weight(P<0.01),a significant amelioration in glycemic control,oral glucose tolerance and lipid metabolism(P<0.01).And oxidative stress and inflammatory responses decreased after oral administration of CSE(P<0.01).CSE also showed protective effects on renal functions,decreasing the ratio of kidney/body weight,24 h urine volume,24 h urine protein,serum creatinine(Scr)and blood urea nitrogen(BUN)(P<0.01).In addition,renal protective effect was also observed in histopathological examination.Immunohistochemical analysis showed that CSE down regulated the expression of RAGE.Histopathological examination of liver showed slight fatty degeneration and inflammation.Histopathological examination of retina showed the protective effects of CSE in retinal morphological changes.Conclusions: With the stable preparation procedure,good quality control and verified anti-diabetic and renoprotective effects in diabetic rats,CSE could be a potential agent in the treatment of type 2 diabetes mellitus(T2DM)and its complications. |
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