The Enterogenous Mechanism Research Of Lindera Alcohol Extract To Protection Acute Alcoholic Liver Injury

Background: Alcoholic liver disease(ALD) is one of the common diseases which cause serious damage to human health, and presents a rising trend year by year. So far there is still no effective therapeutic drugs to be treated at any stage of alcoholic liver disease. So the research of alcoholic liver disease drugs and its mechanism is necessary.Objective: Through the establishment of acute alcoholic liver injury model of rats,observe radix linderae alcohol extract effect on alcoholic liver injury, and then to discuss its enterogenous protection mechanism of alcoholic liver injury.Method: 60 rats were randomly divided into normal control group, model control group, positive control group(0.6 g/kg smecta) and radix linderae alcohol extract group. The radix linderae alcohol extractgroup were divided into 3 groups by dosage: 3g/kg, 2 g/kg and 1 g/kg. The preparation method of the extract of radix linderae alcohol:take medicine appropriately, add 12 bv75 % ethanol to the extract lasting 1.5h, then leach the liquid, and add 10 BV ethanol to the extract lasting 1h, leach the solution again, then merge two liquids, decompress to recover the ethanol and appropriately concentration for the radix linderae alcohol extract. Dilute with suitable amount of purified water before use, make each 1 ml solution is equivalent to 0.5 g crude drugs.(1)After the last drugs delivery to rats metabolic cage, collect the dejecta respectively by group(two pieces per rat), then preserve the total DNA extraction of intestinal flora in-80℃, and take an ERIC- PCR fingerprint detection, and then detect bacterial species by similarity clustering analysis to atlas;(2) Draw blood from hepatic portal vein and abdominal aorta respectively after fasting 12 h over the last drugs delivery, then take a centrifugal preparation of serum to measure the ALT, AST, T.BIL,TNF-α, IL-6, IL-8, NF-κB and ET level;(3)Take a rapid execution to gather the rats’ liver, stomach, large intestine and small intestine 15 cm above ileocecal areaon an glacial table. Preserve the liver tissue respectively in liquid nitrogen and formalin standby, small intestine in 2.5% glutaraldehyde and formalin standby, stomach and large intestine preserved in formalin;Observe each viscera pathological change by HE staining, and take the WB method determination of TLR4 in liver tissue, the expression of CD68, CYP2E1, and observe the change of intestinal mucosa microvilli by electron microscopy.Results: For the acute alcoholic liver injury model of rats, the radix linderae alcohol extract could:1. Reduce serum ALT, AST, TBIL content in serum; 2.Reduce the serum TNF alpha, IL- 6, IL- 8, the nf-kappa B, ET level; 3. Decrease TLR4, CD68, CYP2E1 in liver tissue; 4. Reduce liver tissue pathological damage of the model control rats; 5.Decrease the permeability of the intestinal mucosa; 6. Protect the intestinal microecological bacteria.Conclusion: This study suggests that radix linderae alcohol extract could be acted by regulating intestinal microecological bacteria, to protect the intestinal mucosa, and resume the function of bowel movement, then to reduce leakage of enterogenous endotoxin level; Then reduce the combination of endotoxin and kupffer cell surface receptors, further reduce the activation of the nf-kappa B, eventually reduce the release of inflammatory factors to protect liver.