ABSTRACT
With continuous introduction of relevant national policies on famous classical formulas, the research of famous classical formulas is popular all over the country. Different from other new drugs, in the research and development process of famous classical formulas, substance benchmark is earlier than the product, suggesting that the research and development of substance benchmark is of great significance. Based on previous work of the authors, content of substance benchmark of famous classical formulas was analyzed, which was included in the document <italic>The Management Regulation of Simplified Registration and Approval over Chinese Herbal Medicine Compound Preparations of Ancient Famous Classical Formulas</italic> released by the National Medical Products Administration in May 2018. In this paper, the significance of substance benchmark development was described, a five-stage of research strategy was proposed, covering the prescription textual research and historical evolution, the collection and quality evaluation of medicinal materials, the processing method and quality evaluation of decoction pieces, the preparation and quality research of substance benchmark, the drafting and formulating of quality standard over substance benchmark. At the same time, some suggestions were put forward to the feasibility of compound preparations development over famous classical formulas, the implementation difficulty of resource evaluation over Chinese medicinal materials, and the irrationality on the quality correlation of Chinese medicinal materials. All of these are expected to provide reference and enlightenment for the development and policy officially landed over ancient famous classical formulas.
ABSTRACT
AIM:To explore the effects of genipin (GEN) on high glucose (HG) -induced oxidative stress injury and apoptosis in H9c2 cardiomyocytes.METHODS:H9c2 cells were cultured in vitro and HG-induced injury model was established.H9c2 cells were divided into 4 groups:normal control (NC) group (glucose at 5.6 mmol/L) , HG group (glucose at 50 mmol/L) , NG+GEN group and HG+GEN group.The concentration of genipin was used at 10μmol/L.The viability of the H9c2 cells was measured by CCK-8 assay.The intracellular malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined by enzyme labeling and WST-1 methods, respectively.The activity of lactate dehydrogenase (LDH) in the cell culture supernatant was detected by microplate method.Fluorescent probe DCF was used to detect intracellular levels of reactive oxygen species (ROS).Nucleosome fragments was measured to evaluate cell apoptosis by ELISA.The intracellular mitochondrial membrane potential was detected by JC-1 method.The protein levels of Mn-SOD, cytochrome C (Cyt C) , Bax and cleaved caspase-3 were determined by Western blot.RESULTS:Compared with HG group, the cell viability in HG+GEN group was increased significantly (P<0.05) , the levels of MDA and LDH were decreased (P<0.05) , SOD activity was increased (P<0.05) , the levels of ROS and nucleosome fragments in HG+GEN group were decreased (P<0.05) , and the mitochondrial membranes potential was notably increased (P<0.05).Compared with NG group, the activation of Mn-SOD was decreased, but the protein levels of Cyt C, Bax and cleaved caspase-3 were increased in HG group (P<0.05).Compared with HG group, the activation of Mn-SOD was increased, and the protein levels of Cyt C, Bax and cleaved caspase-3 were decreased in HG+GEN group (P<0.05).CONCLUSION:Genipin protects HG-induced H9c2 cardiomyocytes against oxidative stress injury and apoptosis.