| Epimedii Folium is the dried leaves of Epimedium brevicornum Maxim.,Epimedium sagittatum Maxim.,Epimedium pubescens Maxim.and Epimedium koreanum Nakai.It is a well-known traditional Chinese herbal medicine,which has the functions of "benefiting pneuma,strengthening sinews and bones,tonifying waist and knees,and strengthening heart power".Flavonoids contained in Epimedii Folium are the main bioactive substances,and icariin(ICA)is one of the important components to evaluate the quality of Epimedii Folium medicinal material.Studies have shown that ICA has multiple pharmacological functions,such as anti-oxidation,anti-inflammatory,anti-tumor,immune regulation,neuroprotection,improving cardiovascular function and promoting bone formation.In recent years,extensive scientific research has been conducted on the pharmacological effects of ICA.Cisplatin is a broad-spectrum and efficient anti-tumor drug,which is widely used in the treatment of various types of malignant tumors.Although cisplatin has excellent curative effect on tumor cells,the cardiac and intestinal toxicity caused by cisplatin greatly affects the therapeutic effect of tumors,severely impairs the quality of life of patients,and even threatens the lives of patients,resulting in severe restrictions on its application in clinical oncology.Therefore,there is an urgent need to find a natural drug that does not affect the effect of cisplatin chemotherapy,but also can safely and effectively alleviate its induced cardiac and intestinal toxicity.Previous studies have shown that Epimedii Folium and its main active ingredients have a significant protective effect on the heart,but relatively few studies on intestinal protection.In order to further explore the pharmacological effects of Epimedii Folium and its active ingredients,based on the basic theory of traditional Chinese medicine that "the heart and the small intestine share a paired relationship",this study established the myocardial and intestinal injury model induced by cisplatin through the combination of in vivo and in vitro experiments,and the Epimedii Folium extract(EE)and its main active ingredient ICA were studied systematically,which is fully revealed that EE and ICA could significantly improve cisplatin-induced myocardial and intestinal toxicity,and discussed the possible molecular mechanism of their effect,in order to lay a theoretical foundation for the development of clinical drugs for the protection of chemotherapeutic myocardium and intestinal injury.The main research contents are as follows:1.Exploring the content differences of main chemical components of Epimedii Folium from different producing areas based on fingerprint technologyThis study used HPLC to conduct in-depth research on the flavonoids contained in Epimedii Folium from different producing areas,and confirmed that Epimedii Folium from different producing areas such as Jilin Province,Liaoning Province,Chongqing City and Gansu Province contained Epimedin A,Epimedin B,Epimedin C and Icariin,but the content of each ingredient varied greatly between different producing areas.At the same time,this study further used fingerprint technology to systematically study the chemical constituents and active ingredients of Epimedii Folium medicinal materials from different producing areas,and formed the HPLC fingerprints of Epimedii Folium from different producing areas through multi-point calibration and full peak spectrum matching.According to the similarity and cluster analysis,there was little difference in the chemical compositions and effective components of Epimedii Folium from the same producing areas,and the similarity was mainly concentrated between 0.84～0.99,0.85～0.96 and 0.80～0.95,indicating that there were certain similarities in the chemical compositions and contents between the same producing areas,but there were great differences between different producing areas,which might be related to factors such as the basic source of the medicinal materials,climate,growth environment and harvest time.2.The protective effect of Epimedii Folium extract on cisplatin-induced myocardial and intestinal injuryIn this study,the myocardial and intestinal injury model induced by cisplatin in mice was established to analyze the protective effect of EE on myocardium and intestine from different angles.The results showed that EE could alleviate cisplatin-induced oxidative damage by increasing the activities of glutathione peroxidase(GSH-Px),catalase(CAT)and superoxide dismutase(SOD),and reducing the content of malondialdehyde(MDA).Simultaneous inhibited the secretion of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)and the expression of nuclear factor NF-κB,thereby reducing the inflammatory injury caused by cisplatin.In addition,EE could also inhibit apoptosis by regulating Bax/Bcl-2 and its downstream caspase signaling pathway.EE could improve cisplatin-induced myocardial and intestinal tissue damage by reducing the levels of myocardial injury markers creatine kinase isoenzyme MB(CK-MB),cardiac specific troponin T(cTnT),lactate dehydrogenase(LDH)and intestinal injury marker diamine oxidase(DAO).This study showed that EE had a good protective effect on cisplatin-induced myocardial and intestinal injury.3.The protective effect of icariin on cisplatin-induced cardiomyocyte injuryThe rat myocardial(H9c2)cell injury model induced by cisplatin was established,and ICA was screened by MTT colorimetry to have a good protective effect on cisplatin-induced H9c2 cell injury at doses of 3 μM,6 μM and 12 μM.ICA could significantly reduce the consumption of antioxidants such as GSH-Px,CAT,GSH and SOD by cisplatin,while reduced the level of MDA and regulated the expression of the oxidation regulator SIRT1,indicating that ICA could effectively alleviate the oxidative damage caused by cisplatin to H9c2 cells.ICA reduced cisplatin-induced cardiomyocyte apoptosis by regulating PI3K/Akt,MAPKs and caspase signaling pathways.In addition,ICA could also reduce the occurrence of inflammation by inhibiting the expression of NF-κB.In short,ICA could improve cisplatin-induced H9c2 cell injury by inhibiting ROS-mediated oxidative stress,apoptosis and inflammation.4.The protective effect of icariin on cisplatin-induced intestinal injuryThrough the combination of in vivo and in vitro experiments,this study further explored the improvement effect of ICA on cisplatin-induced intestinal injury and its potential molecular mechanism.ICA could significantly inhibit the consumption of antioxidant enzymes such as GSH-Px,CAT and SOD by cisplatin,while reduced the content of MDA,indicating that ICA could effectively alleviate cisplatin-induced intestinal oxidative damage.Western blot results clarified that ICA could reduce inflammatory damage by regulating the PPARγ/NF-κB/P53 signaling pathway and the expression of classic pro-inflammatory factors such as TNF-α,IL-1β,iNOS and COX-2.The results of immunofluorescence staining and flow cytometry analysis showed that ICA could effectively improve the apoptotic injury caused by cisplatin to the intestinal tract.In addition,ICA played an important role in protecting the intestinal barrier function by up-regulating the expression levels of tight junction proteins ZO-1 and Occludin.The above results proved that ICA could effectively alleviate cisplatin-induced intestinal injury by inhibiting oxidative stress,relieving inflammatory reaction,reducing cell apoptosis and protecting the intestinal barrier function. |
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