| Chronic renal failure(CRF)refers to progressive renal impairment which caused by various primary or secondary Chronic kidney disease(CKD).Renal transplantation surgery and dialysis treatment are often used in the end stage of CRF,but it is difficult to widely promote due to high cost、insufficient renal source、poor cognition and compliance of patient,and different kinds of serious complications.These methods is not suitable for patients with CRF in the early and middle stages.In recent years,traditional Chinese medicine has been widely used in patients with CRF,it shows good results for CRF in early and middle stages especially.Therefore,it has important theoretical research value and practical application potential with the development of traditional Chinese medicine in the treatment of CRF.Ophiocordyceps lanpingensis is a kind of fungus which belongs to the Ophiocordyceps.It is closely related to Ophiocordyceps sinensis.It is a new species of Ophiocordyceps which mainly distributes in Northwest of Yunnan,China,and it was discovered in recent years.Its active ingredients are similar to wild O.sinensis and easy to be cultured artificially.In the O.lanpingensis producing areas of Yunnan province,local minorities have used O.lanpingensis to treat respiratory and urinary diseases for a long time,and its usage is the same as O.sinensis.Compared with O.sinensis,the O.lanpingensis has better characteristics of artificial cultivation and it is easy to large-scale cultivate.Therefore,O.lanpingensis has the potential to replace O.sinensis which to be used as medicine.The aim of this study was to investigate the therapeutic effect of O.lanpingensis powder and its aqueous extract cultured artificially on CRF in mice.It can provide theoretical basis and technical reference for the future artificial culture of O.lanpingensis to replace O.sinensis,and also provide ideas for the prevention and treatment of CRF in clinic.The research of this paper includes the following two aspects:1.Therapeutic effect of aqueous extract of O.lanpingensis on CRF in mice70 C57BL/6 male mice were divided into control group(Normal group),model group(adenine group),losartan positive control group(losartan group),low、medium and high dose of O.lanpingensis powder aqueous extract groups(OLEL,OLEM,OLEH group)and O.lanpingensis powder group(OL group),7 groups with 10 mice in each group.Adenine was given to mice for modeling in experimental group,while distilled water of equal volume was given to the Normal group by gavage.After 30 days of modeling,Losartan solution(10 mg/kg/d)were administered to mice in the positive control group;The OLEL,OLEM and OLEH groups were fed with different doses of aqueous extract of O.lanpingensis powder(0.5 g/kg/d,1.0 g/kg/d,2.0 g/kg/d)and mice in OL group were fed with O.lanpingensis powder(2.0 g/kg/d)respectively,and the Normal group and model group were given 20 mL/kg/d distilled water respectively.On the 7th,14 th and 28 th day after successful modeling,mice in each group were weighed,and eyeballs were taken for blood after the 30 th day of administration.Weighing kidney wet weight after dissection.Serum was separated.Serum urea nitrogen,creatinine,reduced glutathione,phosphorus,malondialdehyde(MDA)、superoxide dismutase(SOD)activity and plasma calcium content were measured.The pathological changes of kidney were observed by HE staining and Masson staining.The O.lanpingensis powder and its aqueous extract can increase the weight of mice、the content of glutathione in serum、the activity of SOD and the content of plasma calcium,and reduce the content of urea nitrogen、creatinine、phosphorus and MDA in mice.In addition,the O.lanpingensis powder and its aqueous extract can significantly improve renal tissue damage and renal interstitial fibrosis in mice with CRF.High,medium and low doses of O.lanpingensis powder aqueous extract can improve CRF in mice,and the high dosage group(2.0 g/kg/d)has the best effect on CRF in mice.The therapeutic effect of the middle dose group(1.0 g/kg/d)group was similar to that of the O.lanpingensis powder group(2.0 g/kg/d).1.0 g O.lanpingensis powder aqueous extract could be collected from 2.0 g O.lanpingensis powder.It can be concluded from the results that O.lanpingensis powder has a good therapeutic effect on CRF which was induced by adenine in mice,and its effective medicinal substances are mainly present in the aqueous extract of O.lanpingensis powder.2.Study on the mechanism of the aqueous extract of O.lanpingensis in the treatment of CRF in miceThe mRNA expression levels of TNF-α,IL-1β,IL-6,MCP-1 and IL-10 and TGF-β were detected by quantitative real-time PCR.The protein expression levels of apoptosis-related proteins Bax,Bcl2,Caspase 3,active-Caspase 3 and Caspase 9,and the protein expression levels of inflammatory-related proteins IL-6,P-65 and P50 were detected by Western blot.The aim of this is to clarify the effect of O.lanpingensis powder and its aqueous extract on the expression level of related proteins in kidney tissue.Kidney was made into paraffin sections,and the apoptotic cells were observed by TUNEL fluorescence staining,and Immunohistochemical method was used to detect the distribution condition of macrophages in tissues,and observe the changes in the number of macrophages.The results show that the O.lanpingensis powder and its aqueous extract could decrease the mRNA expression levels of TNF-α,IL-1β,IL-6,TGF-β and MCP-1,and increased the mRNA expression levels of IL-10.Meanwhile,the O.lanpingensis powder and its aqueous extract can reduce the protein expression levels of Bax,Caspase 3,active-Caspase 3,Caspase 9,P50,IL-6,P-65,and increase the protein expression levels of Bcl2.In addition,the research proves that the O.lanpingensis powder and its aqueous extract could alleviate the apoptosis of cells in kidney tissue,and reduce the number of macrophages in kidney tissue to a certain extent.It can be concluded from the research that the mechanism of O.lanpingensis powder and its aqueous extract in treating CRF in mice is related to inflammation and apoptosis,and it can improve the immune function of mice by regulating macrophages. |
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