BACKGROUND Interdigestive migrating motor complexes(MMC)produce periodic contractions in the gastrointestinal tract,but the exact mechanism of action still remains unclear.Intramuscular interstitial cells of Cajal(ICC...BACKGROUND Interdigestive migrating motor complexes(MMC)produce periodic contractions in the gastrointestinal tract,but the exact mechanism of action still remains unclear.Intramuscular interstitial cells of Cajal(ICC-IM)participate in gastrointestinal hormone and neuromodulation,but the correlation between ICCIM and MMC is also unclear.We found that xiangbinfang granules(XBF)mediated the phase III contraction of MMC.Here,the effects of XBF on gastric antrum motility in W/Wv mice and the effects of ICC-IM on gastric antrum MMC are reported.AIM To observe the effects of ICC-IM on gastric antrum motility and to establish the mechanism of XBF in promoting gastric antrum motility.METHODS The density of c-kit-positive ICC myenteric plexus(ICC-MP)and ICC-IM in the antral muscularis of W/Wv and wild-type(WT)mice was examined by confocal microscopy.The effects of XBF on gastric antrum slow waves in W/Wv and WT mice were recorded by intracellular amplification recording.Micro-strain-gauge force transducers were implanted into the gastric antrum to monitor the MMC and the effect of XBF on gastric antrum motility in conscious W/Wv and WT mice.RESULTS In the gastric antrum of W/Wv mice,c-kit immunoreactivity was significantly reduced,and no ICC-IM network was observed.Spontaneous rhythmic slow waves also appeared in the antrum of W/Wv mice,but the amplitude of the antrum slow wave decreased significantly in W/Wv mice(22.62±2.23 mV vs 2.92±0.52 mV,P<0.0001).MMCs were found in 7 of the 8 WT mice but no complete MMC cycle was found in W/Wv mice.The contractile frequency and amplitude index of the gastric antrum were significantly increased in conscious WT compared to W/Wv mice(frequency,3.53±0.18 cpm vs 1.28±0.12 cpm;amplitude index,23014.26±1798.65 mV·20 min vs 3782.16±407.13 mV·20 min;P<0.0001).XBF depolarized smooth muscle cells of the gastric antrum in WT and W/Wv mice in a dose-dependent manner.Similarly,the gastric antrum motility in WT mice was significantly increased after treatment with XBF 5 mg(P<0.05).Atropine(0.1 mg/kg)blocked the enhancement of XBF in WT and W/Wv mice completely,while tetrodotoxin(0.05 mg/kg)partially inhibited the enhancement by XBF.CONCLUSION ICC-IM participates in the regulation of gastric antrum MMC in mice.XBF induces MMC III-like contractions that enhance gastric antrum motility via ICCIM in mice.展开更多
AIM To investigate the underlying effect of Jianpi Qingchang decoction(JQD) regulating intestinal motility of dextran sulfate sodium(DSS)-induced colitis in mice. METHODS C57BL/6 mice were randomly divided into four g...AIM To investigate the underlying effect of Jianpi Qingchang decoction(JQD) regulating intestinal motility of dextran sulfate sodium(DSS)-induced colitis in mice. METHODS C57BL/6 mice were randomly divided into four groups: the control group, the DSS group, the JQD group, and the 5-aminosalicylic acid group. Except for the control group, colitis was induced in other groups by giving distilled water containing 5% DSS. Seven days after modeling, the mice were administered corresponding drugs intragastrically. The mice were sacrificed on the 15^(th) day. The disease activity index, macroscopic and histopathologic lesions, and ultrastructure of colon interstitial cells of Cajal(ICC) were observed. The levels of tumor necrosis factor-alpha(TNF-α), interleukin(IL)-1β, IL-10 and interferon gamma(IFN-γ), the expression of nuclear factor-kappa B(NF-κB) p65, c-kit, microtubule-associated protein 1 light chain 3(LC3-Ⅱ) and Beclin-l m RNA, and the colonic smooth muscle tension were assessed. RESULTS Acute inflammation occurred in the mice administered DSS. Compared with the control group, the levels of IL-1β, TNF-α, IL-10 and IFN-γ, the expression of LC3-Ⅱ, Beclin-1 and NF-κB p65 m RNA, and the contractile frequency increased(P < 0.05), the expression of c-kit m RNA and the colonic smooth muscle contractile amplitude decreased in the DSS group(P < 0.05). Compared with the DSS group, the levels of IL-10 and IFN-γ, the expression of c-kit m RNA, and the colonic smooth muscle contractile amplitude increased(P < 0.05), the levels of TNF-α and IL-1β, the expression of LC3-Ⅱ, Beclin-1 and NF-κB p65 m RNA, and the contractile frequency decreased in the JQD group(P < 0.05).CONCLUSION JQD can regulate the intestinal motility of DSS-induced colitis in mice through suppressing intestinal inflammatory cascade reaction, reducing autophagy of ICC, and regulating the network path of ICC/smooth muscle cells.展开更多
AIM: To investigate the effects of different parameters of gastric electrical stimulation(GES) on interstitial cells of Cajal(ICCs) and changes in the insulin-like growth factor 1(IGF-1) signal pathway in streptozotoc...AIM: To investigate the effects of different parameters of gastric electrical stimulation(GES) on interstitial cells of Cajal(ICCs) and changes in the insulin-like growth factor 1(IGF-1) signal pathway in streptozotocininduced diabetic rats.METHODS: Male rats were randomized into control, diabetic(DM), diabetic with sham GES(DM + SGES), diabetic with GES1(5.5 cpm, 100 ms, 4 m A)(DM + GES1), diabetic with GES2(5.5 cpm, 300 ms, 4 m A)(DM + GES2) and diabetic with GES3(5.5 cpm, 550 ms, 2 m A)(DM + GES3) groups. The expression levels of c-kit, M-SCF and IGF-1 receptors were evaluated in the gastric antrum using Western blot analysis. The distribution of ICCs was observed using immunolabeling for c-kit, while smooth muscle cells and IGF-1 receptors were identified using α-SMA and IGF-1R antibodies. Serum level of IGF-1 was tested using enzyme-linked immunosorbent assay. RESULTS: Gastric emptying was delayed in the DM group but improved in all GES groups, especially in the GES2 group. The expression levels of c-kit, M-SCF and IGF-1R were decreased in the DM group but increased in all GES groups. More ICCs(c-kit+) and smooth muscle cells(α-SMA+/IGF-1R+) were observed in all GES groups than in the DM group. The average level of IGF-1 in the DM group was markedly decreased, but it was up-regulated in all GES groups, especially in the GES2 group. CONCLUSION: The results suggest that long-pulse GES promotes the regeneration of ICCs. The IGF-1 signaling pathway might be involved in the mechanism underlying this process, which results in improved gastric emptying.展开更多
目的:研究柴胡疏肝散含药血清对体外大鼠胃Cajal间质细胞(interstitial cells of Cajal,ICC)增殖及其细胞周期的影响.方法:体外培养的大鼠胃ICC经荧光鉴定,取生长对数期内的I C C与各含药血清共孵.实验分为空白组、柴胡疏肝散含药血清组...目的:研究柴胡疏肝散含药血清对体外大鼠胃Cajal间质细胞(interstitial cells of Cajal,ICC)增殖及其细胞周期的影响.方法:体外培养的大鼠胃ICC经荧光鉴定,取生长对数期内的I C C与各含药血清共孵.实验分为空白组、柴胡疏肝散含药血清组(5%、10%、20%)及10%多潘立酮含药血清组.通过CCK-8法检测ICC的增殖作用;使用流式细胞术检测ICC细胞周期;Western blot检测ICC细胞周期素依赖性激酶2(cyclin dependent kinase 2,CDK2)、cyclin E的表达.结果:与空白组比较,柴胡疏肝散含药血清各浓度组及10%多潘立酮含药血清组可显著促进ICC增殖(均P<0.01),可提高ICC细胞周期G0/G1期及S期比例(均P<0.01),同时10%柴胡疏肝散及10%多潘立酮含药血清组可显著上调ICC周期蛋白CDK2、cyclin E的表达(均P<0.01).柴胡疏肝散各浓度组内比较,10%、20%浓度组对促进ICC增殖及提高G0/G1期和S期比例均优于5%浓度组(均P<0.05);10%与20%浓度组在促进I C C增殖作用及提高S期比例上差异无统计学意义(均P>0.05).与10%多潘立酮含药血清组比较,10%柴胡疏肝散组对ICC增殖作用及上调CDK2、c y c l i n E的表达均优于10%多潘立酮组(均P<0.05);10%、20%柴胡疏肝散含药血清组提高G0/G1期及S期比例亦优于10%多潘立酮组(均P<0.05).结论:柴胡疏肝散含药血清可提高ICC生长活性,对ICC的增殖分化起促进作用.展开更多
目的探讨内质网自噬在大鼠胃窦Cajal间质细胞(ICCs)内质网应激(ERS)过程中的作用,为改善胃动力障碍提供新靶标。方法采用酶解法分离大鼠胃窦ICCs并行免疫荧光鉴定,使用衣霉素(TM)诱导ICCs构建ERS模型,三甲基腺嘌呤(3-MA)阻断自噬,雷帕霉...目的探讨内质网自噬在大鼠胃窦Cajal间质细胞(ICCs)内质网应激(ERS)过程中的作用,为改善胃动力障碍提供新靶标。方法采用酶解法分离大鼠胃窦ICCs并行免疫荧光鉴定,使用衣霉素(TM)诱导ICCs构建ERS模型,三甲基腺嘌呤(3-MA)阻断自噬,雷帕霉素(Rapa)促进自噬。随机将细胞分为4组并分别给予不同的处理,ICCs组正常培养,ERS组给予2μg/m L TM处理ICCs,3-MA组给予2μg/m L TM和4μmol/m L 3-MA联合作用,Rapa组给予2μg/m L TM和2μmol/m L Rapa联合作用,每组作用时间均为6 h;通过实时荧光定量PCR法检测ERS相关因子GRP78、自噬相关因子LC3B的mRNA表达;CCK-8法检测各组细胞活性。结果成功分离并鉴定ICCs。与ICCs组比较,ERS组、3-MA组和Rapa组的GRP78和LC3B表达均升高(P均<0.05),细胞活性降低(P均<0.05)。与ERS组比较,3-MA组GRP78表达升高(P<0.05),LC3B表达、ICCs细胞活性降低(P均<0.05);Rapa组GRP78表达降低(P<0.05),LC3B表达、ICCs细胞活性升高(P均<0.05)。结论 ICCs的ERS过程伴发内质网自噬,内质网自噬可减轻ICCs的ERS损伤作用。展开更多
文摘BACKGROUND Interdigestive migrating motor complexes(MMC)produce periodic contractions in the gastrointestinal tract,but the exact mechanism of action still remains unclear.Intramuscular interstitial cells of Cajal(ICC-IM)participate in gastrointestinal hormone and neuromodulation,but the correlation between ICCIM and MMC is also unclear.We found that xiangbinfang granules(XBF)mediated the phase III contraction of MMC.Here,the effects of XBF on gastric antrum motility in W/Wv mice and the effects of ICC-IM on gastric antrum MMC are reported.AIM To observe the effects of ICC-IM on gastric antrum motility and to establish the mechanism of XBF in promoting gastric antrum motility.METHODS The density of c-kit-positive ICC myenteric plexus(ICC-MP)and ICC-IM in the antral muscularis of W/Wv and wild-type(WT)mice was examined by confocal microscopy.The effects of XBF on gastric antrum slow waves in W/Wv and WT mice were recorded by intracellular amplification recording.Micro-strain-gauge force transducers were implanted into the gastric antrum to monitor the MMC and the effect of XBF on gastric antrum motility in conscious W/Wv and WT mice.RESULTS In the gastric antrum of W/Wv mice,c-kit immunoreactivity was significantly reduced,and no ICC-IM network was observed.Spontaneous rhythmic slow waves also appeared in the antrum of W/Wv mice,but the amplitude of the antrum slow wave decreased significantly in W/Wv mice(22.62±2.23 mV vs 2.92±0.52 mV,P<0.0001).MMCs were found in 7 of the 8 WT mice but no complete MMC cycle was found in W/Wv mice.The contractile frequency and amplitude index of the gastric antrum were significantly increased in conscious WT compared to W/Wv mice(frequency,3.53±0.18 cpm vs 1.28±0.12 cpm;amplitude index,23014.26±1798.65 mV·20 min vs 3782.16±407.13 mV·20 min;P<0.0001).XBF depolarized smooth muscle cells of the gastric antrum in WT and W/Wv mice in a dose-dependent manner.Similarly,the gastric antrum motility in WT mice was significantly increased after treatment with XBF 5 mg(P<0.05).Atropine(0.1 mg/kg)blocked the enhancement of XBF in WT and W/Wv mice completely,while tetrodotoxin(0.05 mg/kg)partially inhibited the enhancement by XBF.CONCLUSION ICC-IM participates in the regulation of gastric antrum MMC in mice.XBF induces MMC III-like contractions that enhance gastric antrum motility via ICCIM in mice.
基金Supported by the National Natural Science Foundation of China,No.81403355 and No.81573892the Project of 3-Year Action Plan for Shanghai Municipal Chinese Medicine Development,No.ZY3-RCPY-2-2001
文摘AIM To investigate the underlying effect of Jianpi Qingchang decoction(JQD) regulating intestinal motility of dextran sulfate sodium(DSS)-induced colitis in mice. METHODS C57BL/6 mice were randomly divided into four groups: the control group, the DSS group, the JQD group, and the 5-aminosalicylic acid group. Except for the control group, colitis was induced in other groups by giving distilled water containing 5% DSS. Seven days after modeling, the mice were administered corresponding drugs intragastrically. The mice were sacrificed on the 15^(th) day. The disease activity index, macroscopic and histopathologic lesions, and ultrastructure of colon interstitial cells of Cajal(ICC) were observed. The levels of tumor necrosis factor-alpha(TNF-α), interleukin(IL)-1β, IL-10 and interferon gamma(IFN-γ), the expression of nuclear factor-kappa B(NF-κB) p65, c-kit, microtubule-associated protein 1 light chain 3(LC3-Ⅱ) and Beclin-l m RNA, and the colonic smooth muscle tension were assessed. RESULTS Acute inflammation occurred in the mice administered DSS. Compared with the control group, the levels of IL-1β, TNF-α, IL-10 and IFN-γ, the expression of LC3-Ⅱ, Beclin-1 and NF-κB p65 m RNA, and the contractile frequency increased(P < 0.05), the expression of c-kit m RNA and the colonic smooth muscle contractile amplitude decreased in the DSS group(P < 0.05). Compared with the DSS group, the levels of IL-10 and IFN-γ, the expression of c-kit m RNA, and the colonic smooth muscle contractile amplitude increased(P < 0.05), the levels of TNF-α and IL-1β, the expression of LC3-Ⅱ, Beclin-1 and NF-κB p65 m RNA, and the contractile frequency decreased in the JQD group(P < 0.05).CONCLUSION JQD can regulate the intestinal motility of DSS-induced colitis in mice through suppressing intestinal inflammatory cascade reaction, reducing autophagy of ICC, and regulating the network path of ICC/smooth muscle cells.
基金Supported by National Natural Science Foundation of ChinaNo.81270458 and No.81570488
文摘AIM: To investigate the effects of different parameters of gastric electrical stimulation(GES) on interstitial cells of Cajal(ICCs) and changes in the insulin-like growth factor 1(IGF-1) signal pathway in streptozotocininduced diabetic rats.METHODS: Male rats were randomized into control, diabetic(DM), diabetic with sham GES(DM + SGES), diabetic with GES1(5.5 cpm, 100 ms, 4 m A)(DM + GES1), diabetic with GES2(5.5 cpm, 300 ms, 4 m A)(DM + GES2) and diabetic with GES3(5.5 cpm, 550 ms, 2 m A)(DM + GES3) groups. The expression levels of c-kit, M-SCF and IGF-1 receptors were evaluated in the gastric antrum using Western blot analysis. The distribution of ICCs was observed using immunolabeling for c-kit, while smooth muscle cells and IGF-1 receptors were identified using α-SMA and IGF-1R antibodies. Serum level of IGF-1 was tested using enzyme-linked immunosorbent assay. RESULTS: Gastric emptying was delayed in the DM group but improved in all GES groups, especially in the GES2 group. The expression levels of c-kit, M-SCF and IGF-1R were decreased in the DM group but increased in all GES groups. More ICCs(c-kit+) and smooth muscle cells(α-SMA+/IGF-1R+) were observed in all GES groups than in the DM group. The average level of IGF-1 in the DM group was markedly decreased, but it was up-regulated in all GES groups, especially in the GES2 group. CONCLUSION: The results suggest that long-pulse GES promotes the regeneration of ICCs. The IGF-1 signaling pathway might be involved in the mechanism underlying this process, which results in improved gastric emptying.
文摘目的:研究柴胡疏肝散含药血清对体外大鼠胃Cajal间质细胞(interstitial cells of Cajal,ICC)增殖及其细胞周期的影响.方法:体外培养的大鼠胃ICC经荧光鉴定,取生长对数期内的I C C与各含药血清共孵.实验分为空白组、柴胡疏肝散含药血清组(5%、10%、20%)及10%多潘立酮含药血清组.通过CCK-8法检测ICC的增殖作用;使用流式细胞术检测ICC细胞周期;Western blot检测ICC细胞周期素依赖性激酶2(cyclin dependent kinase 2,CDK2)、cyclin E的表达.结果:与空白组比较,柴胡疏肝散含药血清各浓度组及10%多潘立酮含药血清组可显著促进ICC增殖(均P<0.01),可提高ICC细胞周期G0/G1期及S期比例(均P<0.01),同时10%柴胡疏肝散及10%多潘立酮含药血清组可显著上调ICC周期蛋白CDK2、cyclin E的表达(均P<0.01).柴胡疏肝散各浓度组内比较,10%、20%浓度组对促进ICC增殖及提高G0/G1期和S期比例均优于5%浓度组(均P<0.05);10%与20%浓度组在促进I C C增殖作用及提高S期比例上差异无统计学意义(均P>0.05).与10%多潘立酮含药血清组比较,10%柴胡疏肝散组对ICC增殖作用及上调CDK2、c y c l i n E的表达均优于10%多潘立酮组(均P<0.05);10%、20%柴胡疏肝散含药血清组提高G0/G1期及S期比例亦优于10%多潘立酮组(均P<0.05).结论:柴胡疏肝散含药血清可提高ICC生长活性,对ICC的增殖分化起促进作用.
文摘目的探讨内质网自噬在大鼠胃窦Cajal间质细胞(ICCs)内质网应激(ERS)过程中的作用,为改善胃动力障碍提供新靶标。方法采用酶解法分离大鼠胃窦ICCs并行免疫荧光鉴定,使用衣霉素(TM)诱导ICCs构建ERS模型,三甲基腺嘌呤(3-MA)阻断自噬,雷帕霉素(Rapa)促进自噬。随机将细胞分为4组并分别给予不同的处理,ICCs组正常培养,ERS组给予2μg/m L TM处理ICCs,3-MA组给予2μg/m L TM和4μmol/m L 3-MA联合作用,Rapa组给予2μg/m L TM和2μmol/m L Rapa联合作用,每组作用时间均为6 h;通过实时荧光定量PCR法检测ERS相关因子GRP78、自噬相关因子LC3B的mRNA表达;CCK-8法检测各组细胞活性。结果成功分离并鉴定ICCs。与ICCs组比较,ERS组、3-MA组和Rapa组的GRP78和LC3B表达均升高(P均<0.05),细胞活性降低(P均<0.05)。与ERS组比较,3-MA组GRP78表达升高(P<0.05),LC3B表达、ICCs细胞活性降低(P均<0.05);Rapa组GRP78表达降低(P<0.05),LC3B表达、ICCs细胞活性升高(P均<0.05)。结论 ICCs的ERS过程伴发内质网自噬,内质网自噬可减轻ICCs的ERS损伤作用。
