Research Of Cautis Lonicerae Injection Pharmaceutics

White scour of piglet is an recurrent disease in10-30days old piglets whichcaused by escherichia coli with diarrhea and hoar gruel feces; in traditional chineseveterinary medicine, white scour of piglet is because of weakness of the spleen andthe stomach and low resistance. Summer hot and humid poison take advantage of aweak stomach and intestine leading to diarrhea though movement and transformationfailure, unable to distinguish between the clear and the muddy and mixed down. So,on the basis of traditional chinese veterinary medicine and the treatment of practice,we screened out any traditional chinese medicine of clearing away heat and toxicmaterials, antisepsis and anti-inflammation, has foundational ability of prevention andcure white scour of piglet.Caulis Lonicerae as the dry bark of the plant of Caprifoliaceae which has abilityof clearing away heat and toxic materials,dispelling wind and activating collaterals,could cure fever, carbuncle, bloody flux and rheumatism. Caulis Lonicerae has a goodeffect of antisepsis and anti-inflammation, remove swells and activate collaterals. Theorganic acid and iridoid glycoside has obvious biological activities;chlorogenic acidand coffee acid has antibacterial, antiviral effect significantly; chiratin could protectliver function; loganin has anti-inflammatory effects, regulating immune function, etc.Now, the drugs made from Caulis Lonicerae after extraction have been widely used inhuman medicine, Caulis Lonicerae injection could be a new drug pharmaceutics withgood antibacterial anti-inflammatory effects to cure livestock inflammatory diseases.This experiment first make a preliminary study of the quality standards of thepharmaceutics, using TLC to identify the major constituents of chlorogenic acidglycosides and the loganin; using high performance liquid chromatography （HPLC）method for content determination of chlorogenic acid. Then we find chlorogenic acidin0.07¹.75μg had good linear relationship with peak area, the linear regressionequation of the standard curve is y=2.68692e^-007x+2.7590728e^-003, r=0.9999（n=6）, thechlorogenic acid content in the injection is0.84mg/mL.We did the acute toxicity test in mice evaluate injection safety by theintraperitoneal injection of Caulis Lonicerae injection to mice at0.25mL/10g each time, twice each day, continuous observation14days. According to the results, thedrug group mice did not see death,the mental status, appetite, urine,coat the skin,respiratory, eye, nose, mouth and other physiological changes were not see anomaliesin mice; beyond to this, the biggest dosing quantity intraperitoneal injection of CaulisLonicerae injection to mice is0.5mL/10g one day, the Caulis Lonicerae for crude drugcontent is0.5g/10g. The acute toxicity test showed that Caulis Lonicerae injectiondoes not have toxic in this drug concentrations, and has a good security.Owing to the activation of inflammatory signaling proteins inside the cell couldmediate inflammatory cytokine gene transcription and translation, thus enlargeinflammatory injury in the process of inflammation that LPS induced RAW264.7macrophages; therefore, based on the inhibitory effect of proinflammatory cytokinescan reach to a certain extent with the control of inflammation. This experiment byLPS stimulation RAW264.7macrophages to established the vitro models ofinflammation, and used the ELISA method to set-out the production levels of LPSinduced RAW264.7macrophages on TNF-α, IL-6and IL-10by Aulis Loniceraeinjection; our results show, the different mass concentration of Aulis Loniceraeinjection can significantly inhibit the production and release of the proinflammatorycytokine TNF-α and IL-6, and present a dose-dependence; at the same time, thedifferent mass concentration of Aulis Lonicerae injection can rise the productionlevels of LPS induced RAW264.7macrophages on TNF-α, IL-6and IL-10at a dosedependent increase, which point out that Aulis Lonicerae injection have someanti-inflammatory action by regulating the secretion and releases of the cytokines. Inorder to further confirm the anti-inflammatory action mechanism of Caulis Loniceraeinjection, this experiment using Western-blot test method was studied that the effect ofthe LPS induced RAW264.7macrophages IκB-α protein by Caulis Loniceraeinjection, result display, Caulis Lonicerae injection could significantly restrained thephosphorylation of IκB-α, and lighten the degradation level, point out CaulisLonicerae injection could play a part in anti-inflammatory action though regulatingthe activity of IκB-α to control the Production levels of the inflammatory mediator.In summary, in this study we using TLC to identify the major constituents ofchlorogenic acid glycosides and the loganin; using HPLC method for contentdetermination of chlorogenic acid. The acute toxicity test showed that the biggestdosing quantity intraperitoneal injection of Caulis Lonicerae injection to mice is 0.5mL/10g one day, the Caulis Lonicerae for crude drug content is0.5g/10g, CaulisLonicerae injection does not have toxic in this drug concentrations, and has a goodsecurity. ELISA and Western-blot experimental results show that, the CaulisLonicerae injection with a certain anti-inflammatory effects in vitro.