| Purpose: The rat model of liver cirrhosis induced by carbon tetrachloride, to observe the effects of duxiao Ganqing Pill on experimental rat intestinal mucosa cell apoptosis and PI3 K, p-Akt, NF-k B, Ik B protein expression, to explore the duxiao Ganqing Pill on liver cirrhosis in rats intestinal mucosal barrier d ysfunction in pre dry mechanism.Method: The clean level Wistar rats 88 only, to adapt to the environment after 7 days were randomly divided into 6 groups, respectively, 8 rats in the normal group, model group, control group, duxiao Ganqing pill high, medium and low dose group, 16 rats in each. After 12 weeks of 50% carbon tetrachloride solvent rat thigh subcutaneous injection, each 0.5 ml-100g-1, 2- WK-1, and according to the change of body weight adjusted dose, a total of 12 weeks, the preparation of rat model of liver cirrhosis. After the success of the model except the blank group, model group, were given the corresponding medicine orally. Control group: the same period kept under the same conditions, given daily doses of 2ml/100 g of distilled water gavage; model group: the same period kept under the same conditions, the daily given distilled water gavage of 2ml/100g; high dose group(223.4mg / kg): once a day by intragastric administration, continuous administration of 12 weeks; the middle dose group(111.7mg / kg): once a day by intragastric administration, continuous administration of 12 weeks; low dose group(58.9mg / kg): once a day by intragastric administration, continuous administration of 12 weeks; lactulose group(concentration 100g/L, 1ml/100 g by gavage): once a day by intragastric administration, continuous administration in 12 weeks.Result: 1, to observe the general state: duxiao Ganqing pill group and the lactulose group had different degrees of improvement compared with the model group, t he middle dose group was most significant. Middle and high dose groups, better than the control group, but there was still a gap compared with the normal group. The rat mortality in duxiao Ganqing dose group was the lowest of 31.25% pill, 50% in model group, the lactulose group 37.5%, duxiao Ganqing pill low dose group 37.5%, duxiao Ganqing pill low dose group 43.75%.2, effects of duxiao Ganqing Pill on rat liver cirrhosis pathology: normal group showed normal rat liver morphology, structural integrity, clear, liver cells arranged in cords, visible around the central vein radial distribution, hepatic sinusoid is clearly visible, the hepatic lobule structure disappeared in normal model group; lobules of liver, cell volume increased, interstitial lymphocytic infiltration, multinucleate hepatocytes, hepatic steatosis, hyperplasia of fibrous tissue and filled in the liver cells, the formation of false lobules. Failure of low dose group the structure of hepatic lobule, lymphocyte, monocyte infiltration, fibrous tissue hyperplasia, the formation of the interval decreases; the middle dose group showed the structure of hepatic lobule damage, bubble like degeneration of liver cells, inflammatory cells and less visible, even necrosis, fibroplasias, interval decreased, with the model group, but was worse than that of normal group; high dose group, low dose and middle dose group was observed under microscope; lactulose group was similar to model group.3, effects of duxiao Ganqing Pill on liver cirrhosis in rats intestinal tissue pathological changes: normal group showed the intestinal mucosa villi of normal rats; model group of rat intestinal mucosal villus atrophy, shortening, visible fracture, or even visible edema, increased subepithelial space, infiltration of inflammatory cells in lamina propria and muscularis mucosa the epithelium and lamina propria, separation, mucous layer of loose tissue; the treatment group showed intestinal mucosal villi arranged in neat rows, mucous layer becomes compact, narrow gap, epithelia l edema, and reduce the number of inflammatory cells, which in the medium dose group improved the most obvious. Lactulose group also has the above- mentioned improvements, but not so obvious. The score comparison, duxiao Ganqing pill low, medium and high dose group improved the most obvious and better than that of the control group(P < 0.05, P < 0.01); there is no significant difference between the lactulose group and model group(P < 0.05); model group was significantly higher than that of the normal group(P < 0.05); the middle dose group score is still higher than the control group(P < 0.01).4, effects of duxiao Ganqing Pill on liver cirrhosis in rats model of intestinal tissue PI3K/Akt pathway:(1) immunohistochemistry to observe the expression of PI3 K protein in intestinal tissue: the expression of PI3 K protein in the blank control group were, duxiao Ganqing pill low dose group and model group had no significant difference; while the expression of duxiao Ganqing pill dose group, high dose group of PI3 K protein and the lactulose group in intestinal tissue was significantly higher than the model group(P < 0.01); among which duxiao Ganqing dose group was the most obvious in the pill; lactulose group and duxiao Ganqing pill low dose group were similar to the expression(P < 0.05); toxicity removing liver Q ing Wan middle dose and high dose group was higher than that of lactulose group(P < 0.05 or P < 0.01).(2) immunohistochemistry to observe the expression of p-Akt protein in intestinal tissue: the expression of the blank control group were p-Akt, while duxiao Ganqing pill low dose group, high dose of p-Akt group and the lactulose group. The intestinal tissues compared with the model group there is no difference; only duxiao Ganqing pill dose group was significant higher than that of the model group(P < 0.01); lactulose group and duxiao Ganqing pill low dose group and high dose group was similar to the expression(P < 0.05), but significantly lower than duxiao Ganqing dose group(P < 0.01) pill.(3) the expression level of Western blot detection of PI3 K, p-Akt protein, PI3 K protein content: duxiao Ganqing pill low dose group and middle dose group of intestinal tissue PI3 K protein was higher than that in the model group(P < 0.05 or P < 0.01); duxiao Ganqing pill high dose group and the lactulose group and model group had no significant difference; while duxiao Ganqing pill dose group was higher than that of lactulose group(P < 0.01), while the duxiao Ganqing pill low dose group and high dose group and lactulose group. The content of p-Akt protein: duxiao Ganqing pill low, middle dose group and the lactulose group was significantly higher than that in the model group(P < 0.01); duxiao Ganqing pill dose group was higher than that of lactulose group(P < 0.05), while the duxiao Ganqing pill high dose group was lower than that of lactulose group(expression P < 0.05).5, effects of duxiao Ganqing Pill on liver cirrhosis model of intestinal tissue of rats with NF-k B signal pathway:(1) immunohistochemistry to observe the expression of NF-k B protein in intestinal tissue: the expression of NF-k B protein in the blank control group were, and duxiao Ganqing pill low dose group and high dose group and model group had no significant difference(P > 0.05); the expression of NF-k B protein after duxiao Ganqing Wan middle dose group and the lactulose group. The intestinal tissues is lower than the model group(P < 0.05 or P < 0.01), among which duxiao Ganqing dose group was the most obvious pill; duxiao Ganqing content lower than lactulose group dose group was pill(P < 0.05); duxiao Ganqing pill low, high dose group and the lactulose group.(2) immunohistochemistry to observe the expression of Ik B protein in intestinal tissue: the expression of Ik B protein in the blank control group were, duxiao Ganqing pill low dose group and model group had nosignificant difference; the expression of Ik B protein after duxiao Ganqing Wan middle dose group, high dose group and the lactulose group in intestinal tissue was significantly lower than the model group(P < 0.05 or P < 0.01), among which duxiao Ganqing pills in high dose group, the expression of the most obvious; duxiao Ganqing pills, the high dose group was higher than the lactulose group(P < 0.05 or P < 0.01); duxiao Ganqing pill low dose group and the lactulose similar to the group.(3) the expression level of Western blot detection of NF-k B, Ik B protein: NF-k B protein content: duxiao Ganqing pill low, medium, high dose group of NF-k B protein in the intestinal tissue was higher tha n in the model group(P < 0.01), while the lactulose group and model group had no significant difference; duxiao Ganqing pill dose group was higher than that in milk fructose group(P < 0.01), while the duxiao Ganqing pill low, high dose group and the lactulose group. The content of Ik B protein: duxiao Ganqing pill low, medium, high dose group and the lactulose group was significantly higher than that in the model group(P < 0.05 or P < 0.01); duxiao Ganqing pills, the high dose group was higher than the lactulose group(P < 0.01); duxiao Ganqing pill low dose group and lactulose group.6, effects of duxiao Ganqing Pill on intestinal tissue cell apoptosis in rats with liver cirrhosis model: the expression of apoptotic cells was not found in the blank, duxiao Ganqing pill low, medium, high dose group and the lactulose group the number of apoptotic cells was significantly lower than that in the model group(P < 0.01); the elimination of liver Q ing Wan middle dose poisoning group was significantly lower than lactulose group(P < 0.01), the rest of the group and the lactulose group had no significant difference.Conclusion: 1, duxiao Ganqing pill could improve the general state of rats with cirrhosis of the liver, intestinal mucosa morphology and structure to maintain the integrity and the protection of liver function. 2, duxiao Ganqing pill is probably through the up regulation of PI3K/p-Akt signaling pathway, thus inhibiting the apoptosis of intestinal mucosa function, protect the intestinal mucosal barrier. 3, duxiao Ganqing pills can down regulate the expression of NF-k B signaling pathway, so as to reduce the intestinal inflammatory injury, protect the intestinal mucosal barrier. 4, duxiao Ganqing pill has the anti apoptosis effect of intestinal tissue, so as to protect the intestinal mucosal barrier. 5, through theoretical and experimental research, according to the C hinese law founded duxiao Ganqing pill withInvigorating Q i and strengthening the spleen, nourishing yin and clearing heat, purging toxin stagnation to achieve anti- inflammatory, anti apoptosis, protect liver function. |
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