| Objective:In this study, we explored the iNOS gene in obstructive jaundice rats liver injury and its significance; Investigate flavored iNOS expression caused when the Yinchenhao decoction for obstructive jaundice liver injury effect and its protective effect on liver, for clinical reasonable perioperative application of traditional Chinese medicine provides the theory basis.Methods:Male Wistar rat98, Body weight250-300g, animal level for the clean class, were randomly divided into4groups:(1) the control group (group S) had30rats;(2) the obstructive jaundice group (group O)got30ones (3) obstructive jaundice recanalization model group (group OR) got18ones; Obstructive jaundice recanalization model+Chinese medicine lavage group (group ORC) got18ones. The1,3,7,10,14day after the operation,6rats in each group were sacrificed in group O and group S, Group OR and ORC were relieved obstruction obstruction respectively from7days, and1,3,7day after remove obstruction when each phase to be put to death in the rat6, only to return in serum and liver tissue samples. Detection of serum alanine aminotransferase (ALT, AST), serum total bilirubin (TBIL); Another sex in the liver tissue fixed in5%formaldehyde solution, paraffin embedding, line5microns section of hematoxylin-eosin HE staining to observe common pathological changes of liver tissue, and iNOS immunohistochemical staining with the paraffin section, observed the expression of iNOS protein in the liver tissue; Residual liver tissue after liquid nitrogen frozen at80℃refrigerator, rat liver tissue by rt-pcr observation iNOS mRNA expression.Results:1. Experiments showed that the Group S moderately elevated postoperative day1ALT, AST, after gradually decreased to normal range, there was no statistically significant difference; Compared with S group, ALT and AST increased rapidly in group O, the ALT peak on postoperative day1(P<0.05), AST in peak3days postoperatively (P<0.05), after gradually decline,10days after obstruction, ALT and AST rise again; The OR group and remove obstruction of ALT, AST after ORC group were declining, ORC ratio, OR group decreased more significantly (P<0.05). S there was no significant change in TBIL, there was no statistically significant difference; Compared with S group, O group of TBIL after obstruction rapidly increased and peaked at3days (P<0.05), after a slow decline; The OR group and ORC TBIL were significantly declined after remove obstruction, ORC ratio, OR group decreased more significantly (P<0.05).2. Liver tissue pathology change:group S liver cell swelling, necrosis, hepatic cell line order. Group O of visible liver tissue inflammatory cells infiltration, small bile ducts and the lumen of small bile duct hyperplasia, fiber tissue hyperplasia, degeneration necrosis of hepatic cells. Group OR of liver tissue after remove obstruction, with the extension of time, the inflammatory cells infiltration, hyperplasia of fibrous tissue cells significantly reduce, liver cell degeneration necrosis foci. Group ORC of liver cells after remove obstruction pathological changes were better than the group OR.3. RT-PCR method to detect iNOS mRNA expression:group S purpose of iNOS gene expression after surgery is not obvious; Group O, in1-7days, after expressing rises slowly, and group O> group S (P<0.05),further obstruction after7days, began to rise significantly,14days reach the maximum. INOS in group OR compare with group ORC in each phase of the RQ values:After removed obstruction, the expression of iNOS of group OR and group ORC was decreased, and the group ORC> the group OR.(P<0.05)4. Immunohistochemical method to detect the rat liver tissue expression of iNOS: group S postoperative iNOS protein expression is not obvious; Group O, in1-7days, after expressing rises slowly, and group O> group S (P<0.05), further obstruction after7days, began to rise significantly,14days reach the maximum. INOS in group OR compare with group ORC in each phase of the cells:After removed obstruction, the expression of iNOS of group OR and group ORC was decreased, and the group ORC>the group OR, this trend is consistent with that of iNOS gene expression.Conclusions:1. NO has a dual role in obstructive jaundice liver injury. In the early onset, NO synthetic quantity is little, NO can dilate blood vessels, improve liver function. Along With the deterioration of the disease, caused by a variety of stimulating factor increases iNOS expression, generated a lot of NO, too much has NO cytotoxicity, liver function damage in patients with obstructive jaundice.2. Yinchenhao decoction can improve the liver function after obstruction and promote the liver recovery. Compared with the groups not given herbal treatment, ORC group phase when the iNOS mRNA of activation and the expression of iNOS protein were significantly lower than the OR group. Clinical treatment of patients with obstructive jaundice caused by obstruction recanalization after abnormal liver function can use dialectical the prescription, promote the recovery of liver function, reduce the incidence of complications.3. Choose reasonable operation method, combining with perioperative been aid in the treatment of traditional Chinese medicine, can bring good therapeutic effect for clinical. Combine traditional Chinese and western medicine treatment is expected to become the treatment of a new added. |
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