| Subacute ruminal acidosis(SARA) in dairy cows is a common disease in China.The rumen p H was lower than 5.6 or 5.8 for 3 h everyday. The main harm of SARA is reducing dry matter intake, milk yield and income rate, also increasing culling rate and mortality, which makes great harm to the aquaculture industry. The main clinical symptoms of SARA include acute p Hase response, liver abscess and laminitis. The main reason is cows fed the high concentrate of nonstructural-carbohydrate, resulting in the excessive fermentation in the gastrointestinal tract. The decreased rumen p H is caused by a mount of volatile fatty acids(VFA) accumulating in the rumen which can attenuate the barrier function of rumen epithelial. Then the bacterial endotoxin lipopolysaccharide(LPS) translocates from rumen to the body circulation. The increasing LPS can cause the body's immune response, resulting in systemic inflammation, which creates serious harm to the development of aquaculture.Berberine is the main effective component of traditional Chinese medicine Coptis chinensis, which has a long-term history. Due to its broad-spectrum antibacterial effects, it is widely used for many years. Berberine has many p Harmacological and biological effects, such as the antibacterial, anti-inflammation and anti-cancer, which is widely used in the treatment of gastroenteritis and bacterial dysentery. In this study, we induced inflammation model in the rumen epithelial cell by LPS. According to the results of the study, we can prove the protecting effect of berberine in rumen so as to explain the molecular mechanism of the berberine on dairy rumenitis disease.In vivo experiment, we detected the p H, volatile fatty acid(VFA), lactic acid,lipopolysaccharide(LPS) of rumen liquid in healthy cows and calves of subacute ruminal acidosis. We also examined the content of tumor necrosis factor alp Ha(TNF-?), interleukin-1?(IL-1?), interleukin-6(IL-6), lipopolysaccharide(LPS), bloodp Hysiological and biochemical indexes in cow blood. The results showed that p H depressed of SARA cattle in different degree. VFA and lactic acid accumulated in the body and the LPS were released to the blood. The variety of enzymes and ions were affected in vary degrees so as to the normal metabolism of body was affected.In vitro experiment, the calf rumen epithelial cells were collected and cultured.Subacute ruminal acidosis and inflammation of the tumor model was induced by LPS,and then the berberine was used to cure the inflammation. Using enzyme-linked immunosorbent assay(Enzyme-linkedimmunosorbent assay, ELISA) detecting the content of tumor necrosis factor alp Ha(TNF-?), interleukin-1?(IL-1?) and interleukin-6(IL-6) in the cell supernatant. The expression of m RNA in TNF-?,IL-1? and IL-6 were determined by fluorescence quantitative PCR(q RT-PCR). The results showed that berberine significantly reduced TNF-?, IL-1? and IL-6 in the supernatant of rumen epithelial cells. The m RNA expression of TNF-?, IL-1? and IL-6 was deceased in a dose independence. The activation of nuclear factor kappa B(NF-?B), mitogen activated protein kinase(mitogen-activated protein, kinase, MAPK)and toll like receptor 4(TLR4) were detected by Western blot and the nuclear localization of NF-?B was detected by immunocytofluorescence to explain the molecular mechanisms of berberine. The results also indicated that berberine significantly attenuated the activation of NF-?B, MAPK and TLR4 signaling pathway and inhibited the p65 into the nucleus, which further confirmed that subacute ruminal could cause varying degrees of damage to various organs of the body, and berberine treatment can significantly reduce the inflammation of the tumor in a certain extent. |
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