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浙江省宁波市中医院重症医学科
纸质出版日期:2022
移动端阅览
楼屹, 朱之青, 谢莉莉, 等. 电针“足三里”对脓毒症大鼠肠黏膜免疫屏障的影响及机制研究[J]. 针刺研究, 2022,47(5):386-392.
LOU Yi, ZHU Zhi-qing, XIE Li-li, et al. Electroacupuncture at “Zusanli”(ST36) protects intestinal mucosal immune barrier by suppressing apoptosis of intestinal lymphocytes and regulating expression of Bcl-2 and Bax in sepsis rats[J]. Acupuncture research, 2022, 47(5): 386-392.
楼屹, 朱之青, 谢莉莉, 等. 电针“足三里”对脓毒症大鼠肠黏膜免疫屏障的影响及机制研究[J]. 针刺研究, 2022,47(5):386-392. DOI: 10.13702/j.1000-0607.20210580.
LOU Yi, ZHU Zhi-qing, XIE Li-li, et al. Electroacupuncture at “Zusanli”(ST36) protects intestinal mucosal immune barrier by suppressing apoptosis of intestinal lymphocytes and regulating expression of Bcl-2 and Bax in sepsis rats[J]. Acupuncture research, 2022, 47(5): 386-392. DOI: 10.13702/j.1000-0607.20210580.
目的:探讨电针“足三里”对脓毒症大鼠肠黏膜免疫屏障的影响及其可能机制。方法:SD大鼠按随机数字表法分为假手术组6只,模型组、非经非穴组、电针组各15只。以盲肠结扎穿孔法制备脓毒症大鼠模型。电针组大鼠于造模后给予电针双侧“足三里”
非经非穴组造模后给予电针非经非穴处,每次30 min每日1次,连续3 d。观察各组大鼠一般情况及造模后3 d的病死率;检测各组大鼠肠内细菌移位率;HE染色法观察肠黏膜病理形态学变化,TUNEL法检测肠黏膜淋巴细胞凋亡情况,流式细胞术检测CD4
+
和CD8+和CD8
+
T细胞,酶联免疫吸附法检测肠黏膜IL-4和肠黏液sIgA含量,Western blot法检测小肠组织Bcl-2、Bax蛋白表达水平。结果:电针组大鼠病死率为13.33%(2/15)
模型组、非经非穴组均为46.67%(7/15)
电针组较模型组、非经非穴组明显降低(P
<
0.05)。与假手术组比较,模型组肠内细菌移位升高(P
<
0.05)
光镜下肠黏膜病理损伤严重,肠黏膜Chiu评分、淋巴细胞凋亡指数、小肠组织Bax表达均升高(P
<
0.05)
小肠组织CD4+T细胞,酶联免疫吸附法检测肠黏膜IL-4和肠黏液sIgA含量,Western blot法检测小肠组织Bcl-2、Bax蛋白表达水平。结果:电针组大鼠病死率为13.33%(2/15)
模型组、非经非穴组均为46.67%(7/15)
电针组较模型组、非经非穴组明显降低(P
<
0.05)。与假手术组比较,模型组肠内细菌移位升高(P
<
0.05)
光镜下肠黏膜病理损伤严重,肠黏膜Chiu评分、淋巴细胞凋亡指数、小肠组织Bax表达均升高(P
<
0.05)
小肠组织CD4
+
、CD8+、CD8
+
T细胞百分比及肠黏膜IL-4含量、肠黏液sIgA含量、小肠组织Bcl-2表达均明显降低(P
<
0.05)。与模型组及非经非穴组比较,电针组肠内细菌移位率、肠黏膜淋巴细胞凋亡指数、小肠组织Bax表达均降低(P
<
0.05)
小肠组织CD4+T细胞百分比及肠黏膜IL-4含量、肠黏液sIgA含量、小肠组织Bcl-2表达均明显降低(P
<
0.05)。与模型组及非经非穴组比较,电针组肠内细菌移位率、肠黏膜淋巴细胞凋亡指数、小肠组织Bax表达均降低(P
<
0.05)
小肠组织CD4
+
、CD8+、CD8
+
T细胞百分比及肠黏膜IL-4含量、肠黏液sIgA含量、小肠组织Bcl-2表达均升高(P
<
0.05)。结论:电针“足三里”可抑制脓毒症大鼠肠黏膜淋巴细胞凋亡,保护肠黏膜免疫屏障功能,降低肠道通透性,其机制可能与调控Bcl-2家族蛋白有关。Objective To observe the effect of electroacupuncture(EA) at “Zusanli”(ST36) on apoptosis of intestinal T lymphocytes
translocation of intestinal bacteria and expression of intestinal Bcl-2 and Bax proteins and intestinal mucosal immune barrier in sepsis rats
so as to explore its underlying mechanism in relieving sepsis. Methods SD rats were randomly divided into sham operation(n=6)
model(n=15)
non-meridian and non-acupoint(non-acupoint
n=15) and acupoint EA(n=15) groups by using random number table method. The sepsis model was established by using cecal ligation and perforation(CLP) method. EA(2 Hz
2 mA) was applied to bilateral ST36 or non-acupoint for 30 min one hour after modeling
once every day for 3 days. The rats' general conditions and fatality rate in 3 days after modeling were recorded. The liver
spleen and mesenteric lymph nodes were taken for bacterial culture to detect the translocation rate of intestinal bacteria. The small intestinal tissue was taken for observing histopathological changes(Chiu's score: 0-5 points) after HE staining
and for determining the expression levels of Bcl-2 and Bax proteins using Western blot. The intestinal mucosa was sampled for detecting the apop-tosis(apoptotic index) of lymphocytes by using terminal deoxynucleoitidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling(TUNEL) assay
and the counts of CD4+T细胞百分比及肠黏膜IL-4含量、肠黏液sIgA含量、小肠组织Bcl-2表达均升高(P
<
0.05)。结论:电针“足三里”可抑制脓毒症大鼠肠黏膜淋巴细胞凋亡,保护肠黏膜免疫屏障功能,降低肠道通透性,其机制可能与调控Bcl-2家族蛋白有关。
Objective To observe the effect of electroacupuncture(EA) at “Zusanli”(ST36) on apoptosis of intestinal T lymphocytes
translocation of intestinal bacteria and expression of intestinal Bcl-2 and Bax proteins and intestinal mucosal immune barrier in sepsis rats
so as to explore its underlying mechanism in relieving sepsis. Methods SD rats were randomly divided into sham operation(n=6)
model(n=15)
non-meridian and non-acupoint(non-acupoint
n=15) and acupoint EA(n=15) groups by using random number table method. The sepsis model was established by using cecal ligation and perforation(CLP) method. EA(2 Hz
2 mA) was applied to bilateral ST36 or non-acupoint for 30 min one hour after modeling
once every day for 3 days. The rats' general conditions and fatality rate in 3 days after modeling were recorded. The liver
spleen and mesenteric lymph nodes were taken for bacterial culture to detect the translocation rate of intestinal bacteria. The small intestinal tissue was taken for observing histopathological changes(Chiu's score: 0-5 points) after HE staining
and for determining the expression levels of Bcl-2 and Bax proteins using Western blot. The intestinal mucosa was sampled for detecting the apop-tosis(apoptotic index) of lymphocytes by using terminal deoxynucleoitidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling(TUNEL) assay
and the counts of CD4
+
and CD8+ and CD8
+
T cells using flow cytometry. The contents of IL-4 in the small intestine and that of secretory IgA(sIgA) in the small intestinal mucus were determined by using ELISA. Results After modeling
of the 15 rats in each of the 3 groups
7
7 and 2 in the model
non-acupoint and EA groups were dead in the first 3 days
with the fatality rate being 46.67%(7/15)
46.67%(7/15) and 13.33%(2/15)
respectively(being obviously lower in the EA group than in the former two groups
P
<
0.05). Compared with the sham operation group
the incidence of intestinal bacterial translocation
apoptotic index
Chiu's score
and Bax expression were significantly increased(P
<
0.05)
and the percentages of CD4+T cells using flow cytometry. The contents of IL-4 in the small intestine and that of secretory IgA(sIgA) in the small intestinal mucus were determined by using ELISA. Results After modeling
of the 15 rats in each of the 3 groups
7
7 and 2 in the model
non-acupoint and EA groups were dead in the first 3 days
with the fatality rate being 46.67%(7/15)
46.67%(7/15) and 13.33%(2/15)
respectively(being obviously lower in the EA group than in the former two groups
P
<
0.05). Compared with the sham operation group
the incidence of intestinal bacterial translocation
apoptotic index
Chiu's score
and Bax expression were significantly increased(P
<
0.05)
and the percentages of CD4
+
and CD8+ and CD8
+
T cells
IL-4 and sIgA contents and Bcl-2 expression considerably decreased(P
<
0.05) in the model group. In comparison with the model group
modeling-induced increase of incidence of bacterial translocation
apoptotic index and Bax expression
and decrease of percentages of CD4+T cells
IL-4 and sIgA contents and Bcl-2 expression considerably decreased(P
<
0.05) in the model group. In comparison with the model group
modeling-induced increase of incidence of bacterial translocation
apoptotic index and Bax expression
and decrease of percentages of CD4
+
and CD8+ and CD8
+
T cells
IL-4 and sIgA contents and Bcl-2 expression were reversed(P
<
0.05) in the EA group. Conclusion EA at ST36 can reduce death rate and intestinal bacteria translocation incidence in sepsis rats
which may be related to its functions in regulating the expression of intestinal Bcl-2 and Bax proteins and inhibiting the apoptosis of intestinal mucosal T lymphocytes
thereby protecting the immune barrier function of intestinal mucosa to reduce the intestinal permeability.
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