| Objectives To study the influence of the different combinations of compound lizard powders over the expression of Survivin, Bcl-2 and P53 in PLGC(precancerous lesions of gastric cancer) model rats as well as explore the possible mechanism that this preparation acts on PLGC for the purpose of providing an experimental basis for the clinical application of the this preparation in the treatment of PLGC.Methods One week after the adaptive feeding of 120 SPF SD male rats aged 4-6 weeks, 20 were randomized into normal control group, without any treatments, and the remaining 100 were used to build PLGC rats model by the comprehensive modeling method that relies on the 5m L/kg gavage with 0.02 mol/L MNNG solution, irregular diet(2d satiety, 1d hunger), and emotional stimulation(one 10-min tail clamping per week). Eight weeks after, 1 rat randomly chosen from normal control group and 5 modeling rats were killed to prove the consistency with the diagnosis as having PLGC. In the modeling process, 27 rats died, and then 65 rats were randomized into five groups, including(1)model control group(10m L·kg- 1·d- 1 gavage with 0.9﹪ Na Cl solution);(2)80-mesh group of compound lizard powder;(3)100-mesh group of compound lizard powder;(4)balanced mixed group of compound lizard powder with 80 and 100 meshes(XY8 0, XY1 0 0, and XY8 0 ′1 0 0 groups were given 1.5g·kg- 1·d- 1 paste suspension by way of gavage); and(5)vitacoenzyme group(0.3g·kg- 1·d- 1 suspension gavage). There were 13 rats in each group, all of which have been given medicine for 12 consecutive weeks with the purpose of observing their respective situation. At the end of the 12 t h week, all the rats were narcotized and killed after being forbidden to eat and drink for 12 h. Then, the changes in the histopathology of gastric mucosa were observed by eyes, and the routine HE staining test on histopathology and immunohistochemical staining test on adjacent slices were carried out. All these were intended to test the influence of the compound lizard powders over the expression of Survivin and Bcl-2(the apoptosis-inhibiting gene of gastric mucosa) and P53(the apoptosis-promoting gene). Meanwhile, a statistical analysis on the experimental result was performed.Results1. Observation on general situation: with the modeling success rate arriving at 73%, 4 rats from model control group and respective 1 from vitacoenzyme and XY8 0 groups died. In the normal control group, the rats were characterized by obesity, clean and glossy fur, agility, enough food and water intake, granular stool, and ruddy tail. In the model control group, however, the rats were featured with slim body, shaggy, withered and lusterless fur, slow action, less food and water intake, shapeless soft stool, and slightly white tail. For each treatment group, the symptoms above have improved to varying degrees. Before and after treatment, the average weights of the rats in each group were as follows: normal control group(565.54 士 51.70/638.15 士 42.68); model control group(456.23 士35.12/537.22 士 22.92); XY8 0 group(456.54 士 41.32/587.08 士 30.10); XY1 0 0 group(455.08 士 38.56/587.54 士 29.63); XY8 0 ′1 0 0 group(457.54 士 43.03/620.15士 43.44); and vitacoenzyme group(453.31 士 34.07/577.17 士 46.24). According to the statistical result about the weight, the difference between each group and normal control group at the same period before treatment showed statistical significance(P< 0.05); after treatment, all the differences between each group and normal control group, between XY8 0 ′1 0 0 group and vitacoenzyme group, and between XY8 0 and XY1 0 0 groups and XY8 0 ′1 0 0 group at the same period showed s t a t i s t i c a l s i g n i f i c a n c e a s w e l l( P < 0. 0 5), w i t h t h e w e i g h t o f X Y8 0 ′ 1 0 0 g r o u p reaching the highest level.2. Histopathological observation: according to the visual observation on pathomorphology, the normal control group was characterized by(1)the pink gastric mucosa;(2)the plica mucosa with smooth and regular surface; and(3)the thick and extensile gastric wall covered with large amounts of mucus; the model control group showed the characteristics as follows:(1)pale or off-white mucus;(2)disordered plica mucosa;(3)lusterless surface;(4)decrease or disappearance of partial plicae;(5)reduced elastic gastric wall;(6)lack of mucus;(7)hemorrhagic points and particle-like nodes; and(8)dendritic blood vessels under the mucous; the symptoms of the treatment groups have improved to varying degrees, and there was a largest effect on XY8 0 ′ 1 0 0 group. After the HE staining of the slices, microscopic observation showed that the normal control group was equipped with normal gastric mucosa, and the model control group faced the atrophy of gastric mucosa to varying degrees; for all treatment groups, the intestinal metaplasia and dysplasia of gastric mucosa have improved to different degrees, and there was greatest effect on XY8 0 ′1 0 0 group.3. Immunohistochemical result: for Survivin, Bcl-2 and P53, the average optical density(A) of the gastric mucosa of each group was as follows: normal control group(21.46 士 2.69/19.06 士 2.32/19.14 士 2.45); model control group(29.29 士 3.51/30.20 士 4.83/30.16 士 4.82); XY8 0 group(24.18 士 2.82/22.14 士3.57/22.23 士 3.63); XY1 0 0 group(24.05 士 2.80/21.98 士 3.06/22.16 士 3.21); XY8 0 ′1 0 0 group(21.63 士 2.61/19.15 士 2.67/19.35 士 3.24); and vitacoenzyme group(26.59 士 3.36/24.80 士 2.89/25.11 士 3.34). According to the statistical result, the differences between each group and normal control group, between each treatment group and model control group, between each XY treatment group and vitacoenzyme group, and between XY8 0 and XY1 0 0 groups and XY8 0 ′1 0 0 group were statistically significant(P< 0.05), with XY8 0 ′1 0 0 group achieving the lowest A value.4. The results of correlation analysis: according to Pearson correlation analysis, there was a positive correlation between not only the expression of Survivin and that of Bcl-2(P< 0.05, correlation coefficient r=0.935) but also the expression of Survivin and that of P53(P< 0.05, correlation coefficient r=0.899).Conclusion1. The different combinations of compound lizard powders had a positive effect on the fur color, spirit, food and water intake, stool, body weight, activity and survival of PLGC model rats, able to recover the general condition of PLGC model rats.2. The different combinations of compound lizard powders had a positively therapeutic effect on the atrophy, intestinal metaplasia and dysplasia of gastric mucosa of PLGC model rats, able to reverse PLGC effectively.3. The different combinations of compound lizard powders could reduce the expression of Survivin and Bcl-2(apoptosis-inhibiting gene) as well as the expression of P53(apoptosis-promoting gene).4. The positive correlation between Survivin and Bcl-2 as well as P53 suggested that there was a synergistic effect between Survivin and Bcl-2 as well as P53 in the occurrence and development of PLGC.5. The different combinations of compound lizard powders could further reverse PLGC with its unique prescription based on the balanced mix of 80 meshes and 100 meshes as well as paste drug delivery. The action mechanism was likely to down-regulate the expression of Survivin and Bcl-2(apoptosis-inhibiting gene) and reduce the mutation of P53(apoptosis-promoting gene) so as to intervene in PLGC and delay canceration. |
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