ObjectiveTo investigate the effects of Aucklandia and Coptis pills on cellular apoptosis and the expression of Bcl-2 and Bax mRNA in model rats with ulcerative colitis (UC). MethodsFifty male Wistar rats at the age of seven weeks were randomly divided into five groups: control group, model group, Chinese medicine group (Aucklandia and Coptis pills), inhibitor group, and Chinese medicine plus inhibitor group. The experiment was performed with rats of UC induced by trinitro-benzene-sulfonic acid enema. AG-490 and Aucklandia and Coptis pills were administrated to them by intraperitoneal injection and gavage. Colonic mucosal tissues in rats of all the five groups were observed and evaluated by light microscope. Cellular apoptosis in colonic mucosal tissues was detected by TUNEL. The expressions of Bcl-2 and Bax mRNA were detected by reverse transcription polymerase chain reaction. ResultsThere were many ulcers in the colon of UC rats, and pathological changes in colonic mucosa such as inflammation, congestion and edema were observed in the colon of UC model rats by naked eye and microscope. Compared with UC rats without treatment, Aucklandia and Coptis pills alleviated colonic mucosal injuries and decreased apoptosis rate of colonic epithelial cells, while the expression of Bax mRNA was decreased in the colonic mucosa in UC rats treated with Aucklandia and Coptis pills, and Bcl-2 mRNA expression was increased. ConclusionAucklandia and Coptis pills can effectively inhibit mRNA expression of apoptosisrelated molecules to down-regulate colonic epithelial cells apoptosis in colonic mucosa in rats with UC.
To detect the cell density, apoptotic incidence and the expressions of Bax and Caspase-3in human lumbar intervertebral discs, so as to further understand the mechanism of human lumbar intervertebral discdegeneration and provide a new idea for biologic treatment of it in future. Methods From May to December in 2006,30 human lumbar intervertebral discs in experimental group(L2 to S1)were surgically collected from 27 patients undergoing posterior lumbar intervertebral discoidectomy and fusion. All the cases were affirmed by MRI and they never experienced discography, collagenolysis of nucleus pulposus and percutaneous laser disc decompression. The control group consisted of 20 human lumbar intervertebral discs(L2 to S1)harvested from 5 young men without spine-related condition immediately after their accidental death. Apoptotic disc cells were detected by TUNEL and histomorphology, and immunohistochemical staining with SP method was performed to examine the expressions of Bax and Caspase-3 in all specimens. Results HE staining disclosed that the average cell density in control group (17.16 ± 1.22)/HP was higher than that in experimental group (12.41 ± 0.95)/HP (P lt; 0.01). However, TUNEL staining observed that the average TUNEL positive incidence in control group (6.97% ± 0.92%) was lower than that in experimental group (12.59% ± 0.95%), (P lt; 0.01). Immunohistochemical staining with SP method showed that the Bax and Caspase-3 positive incidence of nucleus pulposus in control group (11.02% ± 1.18%, 9.01% ± 1.00%) were lower than those in experimental group (19.29% ± 1.18%, 15.07% ± 0.97%), (P lt; 0.01). The results of the average gray scale value of nucleus pulposus in control group were 187.33 ± 7.88 and 185.68 ± 3.26, respectively, with 124.98 ±6.69 and 160.13 ± 4.37 in experimental group. There was significant difference between the two groups (P lt; 0.01). When thetotal 50 specimens in the two groups were analyzed, TUNEL positive incidence showed significant inverse correlations with their respectively corresponding cell densities (r = － 0.88, r = － 0.93, P lt; 0.01). The Bax and Caspase-3 positive incidence of nucleus pulposus showed significant positive correlation with the TUNEL positive incidence of nucleus pulposus (r = 0.83, r = 0.91, P lt; 0.01). Conclusion The decrease of cell density is involved in the development of human lumbar intervertebral disc degeneration. Bax and Caspase-3 might play a role in disc cell apoptosis in nucleus pulposus of human lumbar intervertebral disc.
Objective To investigate the expressions of heat shock protein 27 (HSP27), Bcl-2, and Bax proteins of the nerve cells after spinal cord ischemia/reperfusion injury (SCII) in rats and their relationship. Methods Seventy adult male Sprague Dawley rats (weighing, 200-220 g) were randomly divided into the sham operated group (sham group, n=35) and the SCII group (n=35). Only the left renal artery was exposed with no occlusion of the abdominal aorta in the rats of sham group. The left renal artery was exposed with occlusion of the abdominal aorta for 20 minutes in the rats of SCII group. At 4, 8, and 12 hours and at 1, 2, 3, and 5 days, reperfusion treatment was performed in 5 rats respectively, and then the spinal cord tissue was harvested to detect the expressions of HSP27, Bcl-2, and Bax protein of the nerve cells by using immunohistochemistry staining. Results The HSP27 began to express at 4 hours, reached the peak at 3 days, and decreased at 5 days in SCII group; significant differences were found between at 3 and 5 days and at the other time points (P lt; 0.05). The Bcl-2 expression increased at 4 hours, reached the peak at 1 day and maintained a high level at 2 days, and then gradually decreased; significant differences were found between at 1 and 2 days and at the other time points (P lt; 0.05). The Bax expression reached the peak at 12 hours and 3 days, and decreased at 5 days; significant differences were found between at 12 hours and 3 days and at the other time points (P lt; 0.05). A little expression of each protein was observed in sham group at different time points; the expressions of HSP27, Bcl-2, and Bax proteins in SCII group were significantly higher than those in sham group at different time points (P lt; 0.05). Conclusion There may be the time window of self repair after SCII. High expression of HSP27 has an obvious protective effect on the SCII in rat, by promoting the expression of the anti-apoptotic protein Bcl-2 and reducing the expression of the pro-apoptotic protein Bax so as to inhibit spinal cord cell apoptosis.
ObjectiveTo detect the expression of Notch1, Bax, Bcl-2 genes in rat knee joint cartilage cells in a state of activation and inactivation of the Notch signaling pathway, and preliminarily study the mechanism of Notch signaling pathway on experimental rat knee osteoarthritis (OA) chondrocytes apoptosis.MethodsA total of 34 specefic-pathogen-free Sprague Dawley rats were selected, of which 32 were established the right knee OA models using Hulth method, and the other 2 were normally fed. Four weeks later, two randomly selected OA rats and the two normally fed rats were put to death, to observe the morphological changes of the right knee and ensure the OA models were successfully established by pathology examination. The remaining 30 rats were randomly divided into three groups with 10 in each. The rats were injected intra-articularly on each Tuesday and Friday, with Nocth signal pathway specific activator Jagged1 protein (25 ng/kg) in the activation group, γ-secretase inhibitor DAPT (GSI-IX) (100 ng/kg) in the inhibition group, and phosphate-buffered saline in the control group, respectively. The rats were sacrificed after 8 weeks of articular cavity injection. Taking the right knee articular cartilage speciments of femoral condyle, we observed the degeneration of articular cartilage of the three groups, observed the histomorphological changes by microscope, evaluated the Mankin scores, and used the immunohistochemistry to detect the expression of Notch1, Bax, Bcl-2 proteins.ResultsAfter the 8-week articular cavity injection, the Mankin scores in the activation group, the inhibition group, and the control group were 3.40±0.84, 6.70±0.95, 11.10±1.37, respectively, and the differences between the three groups were statistically significant (P<0.05). The positive rates of Notch1 and Bax of chondrocyte in the inhibition group were lower than those in the control group and the activation group (P<0.05), while the positive rate of Bcl-2 of chondrocyte in the inhibition group was higher than that in the control group and the activation group (P<0.05).ConclusionActivating the Notch signaling pathway may facilitate the chondrocyte apoptosis and aggravate OA by up-regulating Bax protein expression and down-regulating Bcl-2 protein expression; inhibiting the Notch signaling pathway may inhibit the chondrocyte apoptosis and relieve OA by up-regulating Bcl-2 protein expression and down-regulating Bax protein expression.
Objectives To investigate the expressions and significance of E2F1, ID1, and Bax protein in gallbladder adenocarcinoma tissues. MethodsThe expressions of E2F1, ID1, and Bax protein in 70 cases of gallbladder adenocarcinoma, 20 cases of high level intraepithelial neoplasia, 30 cases of low level intraepithelial neoplasia, and 20 cases of cholecystitis tissues were tested by using immunohistochemical method. ResultsThe positive expression rates of E2F1, ID1, and Bax protein in gallbladder adenocarcinoma was 84.3%, 70.0%, and 25.7%, respectively; the positive expression rates in high level intraepithelial neoplasia was 75.0%, 65.0%, and 55.0%, respectively; the positive expression rates in low level intraepithelial neoplasia was 16.7%, 23.3%, and 56.7%, respectively; and the positive expression rates in cholecystitis tissues was 10.0%, 20.0%, and 75%, respectively.The positive expression rates of E2F1 and ID1 protein in gallbladder adenocarcinoma were significantly higher than those intraepithelial neoplasia and cholecystitis tissues (P < 0.05), but the positive expression rate of Bax protein in gallbladder adenocarcinoma was lower (P < 0.05).The expressions of E2F1 and ID1 protein were significantly correlated with clinical Nevin staging of gallbladder adenocarcinoma (P < 0.05), but not correlated with the gallbladder adenocarcinoma differentiation degree (P > 0.05).The expression of Bax protein was related to the gallbladder adenocarcinoma differentiation degree (P < 0.05), but not correlated with clinical Nevin staging (P > 0.05).The expression of E2F1 protein was negatively correlated with expression of Bax protein (r=-0.375, P < 0.05), ID1 protein expression has nothing to do with the protein expression of Bax protein (P > 0.05).The expression of E2F1 protein was positively correlated with ID1 protein (r=7.031, P < 0.05). ConclusionsThe E2F1, ID1, and Bax may play an important role in the generation and development of the gallbladder adenocarcinoma.The combined detection of E2F1, ID1, and Bax have important guiding significance for auxiliary diagnosis and clinical staging of gallbladder adenocarcinoma.
ObjectiveTo study the expression of apoptosissuppressing gene (bcl2) and apoptosispromoting gene (bax) in colorectal cancerous tissue and transitional mucosas. MethodsColorectal cancerous tissue, transitional mucosas (3 cm from the cancerous tissue) and normal tissue were taken respectively in thirtyone cases. Immunohistochemical technique SP method was used to detect the expression in those tissues. ResultsThe positive expression rate of bcl2 protein in cancerous tissue and transitional mucosa were 64.5%and 60.0% respectively and significantly higher than that in normal tissue (P＜0.05). The positive expression rate of bcl2 protein in normal tissue was 35.0%. The positive expression rate of bax protein in cancerous tissue and transitional mucosa were 45.2%and 45.0% respectively and significantly lower than that in normal mucosa (P＜0.05). The positive expression rate of bax protein in normal tissue was 60.0%. There was no obvious difference in the positive rate of bax and bcl2 protein between cancerous tissue and transitional mucosa (Pgt;0.05). The expression rate of bax and bcl2 protein in colorectal cancer was irrelative to clinical pathological gradation and clinical stage (Pgt;0.05). ConclusionThere is over expression of bcl2 protein and low expression of bax protein in colorectal cancer and transitional mucosa. bcl2 protein and bax protein can affect the generation of colorectal cancer by participating in the regulation of apoptosis. But it is irrelative to clinical pathological gradation and clinical stage. Transitional mucosas should be viewed as precancerous lesion and resected during operation.
摘要:目的:動態觀察大鼠腦出血后血腫周圍組織補體激活與細胞凋亡的規律。方法:用膠原酶注入到大鼠尾狀核的方法制作腦出血模型。將大鼠分為腦出血、假手術組、正常組3組。采用蘇木素伊紅(HE) 染色、免疫組織化學染色及原位末端脫氧核苷酸轉移酶介導的dUTP 缺口末端標記法(TUNEL)分別觀察各組在腦出血后第6 h、12 h、24 h、48 h、72 h、5 d、7 d時血腫周圍補體C3、促凋亡基因(Bax)、抑凋亡基因(Bclxl)及TUNEL的表達。結果補體C3的表達峰值在24~48 h;TUNEL、Bax蛋白表達術后12h增加,48~72 h達高峰,而Bclxl蛋白表達高峰在48h。結論:大鼠腦出血后血腫周圍組織補體C3的表達增加與細胞凋亡的演變趨勢一致,C3與凋亡有相關。Abstract: Objective: To study the complement activation and apoptosis regular genes changes in the tissues of the perihematoma of intracerebral hemorrhage (ICH) in rats. Methods: Intracerebral hemorrhage was induced in rats by injection of bacterial collagenase into the caudate nucleus. Histopathological changes were studied in 6 h,12 h, 24 h, 2 d, 3 d, 5 d, 7 d after the injection. The immunohistochemistry and TUNEL analysis were performed. The expression of complement factor C3, the TUNELpositive cells, the proapoptotic gene expression (Bax) and the antiapoptotic gene (Bclxl) were examined. Results: The expression of C3 increased to its maximum between 2448 h. The TUNELpositive cells and Bax protein expression increased gradually and reached the peak level between 4872 h. The Bclxl protein reached the peak level at 48 h. The correlation analysis showed that the quantity of C3 was positively related to that of the TUNELpositive cells, but the bax protein was not related to Bclxl protein. Conclusion: The expression of complement factor C3 may contributes to the nerve injury after cerebral hemorrhage and relate to the apotosis in the tissues surrounding the hametoma in rats.