The Pharmaceutical Research Of Anti-intestinal Endotoxemia Drug-Shuangligan

Intestinal endotoxemia (IETM) plays an important role in the occurrence and evolvement of all kinds of liver diseases. IETM stimulates Kupffer cells to release cytokines, inflammatory mediators, free radicles, nitric oxide(NO) and endothelin-1 (ET-1) that are considered as main factors to result in liver injury, inflammation, hepatic microcirculatory disturbance. Shuangligan(SLG), a herbal preparation, composed of Radix et Rhizama Rhei and Radix et Rhizama Salviae Miltiorrhiza was the research outcome of the Institute of Hepatology at the Shanxi Medical University over twenty years. Both laboratorial research and clinical observation showed that SLG was evident in inhibiting IETM. It was proved that SLG can decrease the level of endotoxin, improve the function of cellular immunoresponse, obviously reduce the content of HBV DNA and lower transaminase in the hepatitis patients. The comprehensive pharmaceutical research on SLG helped to clarify its mechanism of anti-liver injury and lead a new drug become possible in the no longer future. The research is composed of three sections: Section one The study for quality standard of SLG Section two The extraction and separation of the effective components in SLG Section three The study for the effect of effective components in SLG on anti-acute liver injury Section one The study for quality standard of SLG Object: To set up a method to measure quantitatively the effective components in SLG deriving from Radix et Rhizoma and Radix et Salviae Miltiorrhiza to monitor and ensure the quality of SLG., which play a key role to apply the drug safely , effectively and rationally. Experiment one Establishing a method to determine Tanshinone ⅡA in SLG quantitatively. Tanshinone ⅡA was one of the effective components deriving from Radix et Salviae Miltiorrhiza. Through determinating the content of Tanshinone ⅡA, the quality of SLG was monitored. Method: ①Proper methods were used to prepare the control article solution of Tanshinone ⅡA, the test article solution of SLG and the test article solution of Radix Salviae Miltiorrhiza. ②The UV detection wavelength of Tanshinone ⅡA for HPLC was determined through UV-scanning. ③The chromatographic condition of HPLC for Tanshinone ⅡA was studied. The proper condition with good separation and without obvious interference was selected. ④The linear equation between the absorption peak area and the concentration of Tanshinone ⅡA was obtained according to the peak area of gradient concentration of Tanshinone ⅡA . ⑤The accuraracy and credibility of instrumental analysis result were investigated through the precision experiment and the ruddesness experiment. ⑥The correctness of the experimental result was checked through measuring the content of Tanshinone ⅡA in SLG in different moment. ⑦The recovery test of SLG test article solution was determined and the content of Tanshinone ⅡA in SLG was investigated on the experimental condition. ⑧T he quality standard of SLG at Tanshinone ⅡA content was established by measuring the SLG content in different blocks. Results: The HPLC analysis was carried out on a column of TIANHE-ODS (250mm×4.6mm i.d), using a mobile phase of methanol:water (78:22) with a flow rate of 1.0ml/min. The wavelength of detector was 270nm and the column temperature was 40℃. A accurate and reliable method was setup to determine the content of Tanshinone ⅡA in SLG . The standard of SLG was established, in which the content of Tanshinone ⅡA was no less than 0.39mg /g. Experiment two Establishing a method for the quantitative determination of Danshensu in SLG Danshensu(DSS) was one of the effective components deriving from Rhizama Salviae Miltiorrhiza , so it was selected as one of indexes to monitor the quality of SLG. Method: The method for the quantitative determination of Danshensu through HPLC was built as Experiment one shown. Results: The HPLC analysis was carried out on a column of TIANHE-ODS (250mm×4.6mm i.d), using a mobile phase of methanol:0.5% acetic acid (13:87) with a flow rate o...