1.Expression of LRP16 in human endometrial carcinoma tissues and its role on cell proliferation
Yujiao ZHANG ; Ling LIU ; Chunyan WANG ; Ping ZHAN ; Zonghui WU ; Wen HE ; Dingyu WANG
Chongqing Medicine 2013;(25):2983-2985
Objective To investigate the expression change of LRP16 in endometrial cancer tissues and its influence on the pro-liferation of human endometrial carcinoma HEC-1-B cells .Methods HEC-1-B cells were transfected with LRP16 .RT-PCR was used to examine the expression of LRP16 in 26 normal endometrium specimens ,10 endometrial cancer specimens .RT-PCR was used for verifying the transfection success .WES-T was used to observe the proliferation change of HEC-1-B cells .Results The positive expression rate and level of LRP16 mRNA in the endometrial cancer tissues were 83 .33% and 0 .82 ± 0 .21 ,which were significantly higher than 30 .00% ,0 .47 ± 0 .18 in the normal endometrium tissues(P<0 .05) .The RT-PCR detection results revealed that the expression of LRP16 mRNA after transfection was significantly increased .HEC-1-B cells in the transfection group could continued to proliferate in vitro ,but the proliferation capacity was not increased .Conclusion The expression abnormality of LRP16 may be closely related to the occurrence and progress of endometrial cancer ,LRP16 gene may have potential value for the endometrial canc-er gene therapy .
2.Electroacupuncture delays articular cartilage degeneration in osteoarthritisvia Ras-Raf-MEK1/2-ERK1/2 signaling pathway
Changlong FU ; Houhuang CHEN ; Dingyu ZHU ; Zhuile WU ; Xin XU ; Chunsong ZHENG ; Li LI ; Xianxiang LIU ; Xihai LI ; Mingxia WU
Chinese Journal of Tissue Engineering Research 2017;21(24):3790-3795
BACKGROUND:Previous studies have found that electroacupuncture can delay articular cartilage degeneration mediated by JAK-STAT signaling pathway through upregulating the expression level of transforming growth factor β1 as well as mRNA expression levels of STAT3, Smad3 and LepR. In the meanwhile, electroacupuncture can inhibit the mRNA expression of p38 and Fas mRNA mediated by MAPK signaling pathways, further inhibiting the apoptosis of chondrocytes. OBJECTIVE: To explore the effect of electroacupuncture on the degeneration of articular cartilage in rats with knee osteoarthritis based on Ras-Raf-MEK1/2-ERK1/2 signaling pathway. METHODS:120 male healthy Sprague-Dawley rats aged 2 months olds were selected and randomly divided into normal, model, 15-minite electroacupuncture and 30-minute electroacupuncture groups (n=30 per group). The rats in the latter three groups received the intra-articular injection of 4% papain bilaterally, and the remaining rats received no intervention. At 2 weeks after modeling, the latter two groups were respectively given 15- and 30-minute electroacupuncture, five times weekly for consecutive 12 weeks. The morphology of the cartilage was observed by hematoxylin-eosin staining, the expression level of interleukin-1β in the synovium was detected by ELISA assay, and the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were detected by western blot analysis. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that: in the model group, the cartilage surface was rough, the cartilage layer became thinner, and the cartilage structure was damaged with incomplete tidal line; in the 15- and 30-minute electroacupuncture groups, the cartilage structure was complete with clear layers and complete tidal line. ELISA showed that the expression level of interleukin-1β in the model group was significantly higher than that in the normal group (P< 0.01), and the level in the 15- and 30-minute electroacupuncture groups was significantly lower than that in the model group (P < 0.05). Western blot assay found that compared with the normal group, the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were increased in the model group. However, all above protein levels except ERK1/2 in the 15- and 30-minute electroacupuncture groups were significantly lower than those in the model group (P < 0.01,P < 0.05). To conclude, electroacupuncture inhibits the degeneration of articular cartilage in osteoarthritisvia Ras-Raf-MEK1/2-ERK1/2 signaling pathway and downregulating the expression level of interleukin-1β.
3.Regulatory effect of human umbilical cord mesenchymal stem cells on intestinal barrier function in diabetic nephropathy rats
Yaru WU ; Yan MI ; Kaiyue WEI ; Heping GAO ; Dingyu ZHANG ; Caili WANG
Chinese Journal of Tissue Engineering Research 2024;28(19):2967-2973
BACKGROUND:Diabetic nephropathy is an important cause of end-stage renal disease,and intestinal barrier damage plays an important role in the occurrence and development of diabetic nephropathy. OBJECTIVE:To observe the protective effect of human umbilical cord mesenchymal stem cells on the intestinal barrier in rats with diabetic nephropathy. METHODS:Thirty 8-week-old male SD rats were randomly assigned to healthy control group,model group and human umbilical cord mesenchymal stem cell group,with 10 rats in each group.Rats in the human umbilical cord mesenchymal stem cell group were injected with 1×106 human umbilical cord mesenchymal stem cells through the tail vein once a week for 4 weeks after the model establishment of diabetic nephropathy.Rats in the healthy control group and the model group were injected with an equal volume of PBS at the same time.1 week after the last injection,the histomorphological changes in the kidney and colon were observed under a light microscope.The expressions of ZO-1 and Occludin in the colon tissue of rats were detected by immunohistochemistry.Serum D-lactic acid and lipopolysaccharide levels were detected by ELISA.In addition,the distribution of human umbilical cord mesenchymal stem cells labeled with DiR dye in rats was observed by in vivo imaging system.The expression of human mesenchymal stem cell surface marker antigens CD44 and CD90 in colon tissue was detected by immunohistochemistry. RESULTS AND CONCLUSION:(1)Compared with the model group,human umbilical cord mesenchymal stem cell transplantation significantly inhibited the increase of urea nitrogen,serum creatinine,24-hour urine protein level and urinary albumin/creatinine ratio in diabetic nephropathy rats(all P<0.05).(2)The expression of human mesenchymal stem cell surface markers CD44 and CD90 was found in the colon of diabetic nephropathy rats.(3)Compared with the healthy control group,the expression levels of tight junction proteins Occludin and ZO-1 in the colon tissue of the model group were significantly reduced,while the expressions of Occludin and ZO-1 were significantly increased after treatment with human umbilical cord mesenchymal stem cells.(4)Compared with the model group,human umbilical cord mesenchymal stem cell transplantation significantly reduced serum D-lactic acid and lipopolysaccharide levels in diabetic nephropathy rats.(5)The results suggest that human umbilical cord mesenchymal stem cells may protect the intestinal barrier function by enhancing the expression of intestinal tight junction proteins in diabetic nephropathy rats.
4.Study on the Mechanism of Periplaneta americana Extract Reversing Multi-drug Resistance of Human Hepato- cellular Carcinoma HepG 2/ADM Cells
Cailin LI ; Dingyu WU ; Hong LYU ; Honghan ZHANG ; Yanquan WANG ; Mohammerd NAJIB ; Fang PENG
China Pharmacy 2020;31(15):1816-1823
OBJECTIVE:To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3(shorted for “degreasing cream ”and“CⅡ-3”)reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS :MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib (positive control ),degreasing cream and C Ⅱ-3 on HepG2/ADM cells ,then IC 20 was calculated. The experiment was divided into sensitivity drug ,drug-resistance group ,sorafenib group,degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ,and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ,and other 3 groups were treated with relevant medicine (IC20 as drug concentration ). The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp (expression produce of MDR1 gene),LRP,BCRP] and that related to enzyme-mediated multidrug resistance pathway (GST-π and Topo Ⅱ). RESULTS :The IC 20 of degreasing cream ,CⅡ-3 and sorafenib were (2.40±0.16), (200.44±27.52),(18.00±1.82)μg/mL,respectively. Compared with sensitivity group ,the protein expressions of Bcl- 2,P-gp, LRP,BCRP and Topo Ⅱ,the mRNA expressions of MDR 1, LRP,BCRP and GST-π were increased significantly in drug resistance group (P<0.05 or P<0.01). Compared with @qq.com drug-resistance group ,the mRNA and protein expression of MDR1 mRNA and LRP ,BCRP,GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group(P< 0.05 or P<0.01);the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased (P<0.01), while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group (P<0.05). CONCLUSIONS:Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux , promoting cell apoptosis ,reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.
5.Effect of joss stick moxibustion combined with pricking and cupping for acute herpes zoster and its mechanism of analgesia.
Guoping YE ; Meiling SU ; Dingyu ZHU ; Linyun ZHANG ; Wang LIN ; Li HUANG ; Mingxia WU
Chinese Acupuncture & Moxibustion 2017;37(12):1289-1293
OBJECTIVETo observe the effects of conventional western medication and joss stick moxibustion combined with pricking and cupping for herpes zoster in acute stage, and to explore its analgesic mechanism.
METHODSSeventy patients with acute herpes zoster were randomized into an observation group (33 cases after 2 dropping) and a control group (34 cases after 1 dropping). Patients in the observation group were treated with joss stick moxibustion combined with pricking and cupping at localpoints for 7 times, once every other day. Oral acyclovir, vitamin Band mecobalamin tablets were applied in the control group for continuous 14 days, and interferon injection was used for continuous 6 days, etc. The herpes evaluation indexes of blister stopping time, scab time and decrustation time as well as pain intensity were observed before and after treatment. Peripheral serum substance P (SP) content of herpes local situation was detected. The comprehensive effects were evaluated.
RESULTSThe blister stopping time, scab time and decrustation time in the observation group were shorter than those in the control group (all<0.05). There was no statistical significance for pain relief degree between the two groups (>0.05). The pain beginning to ease time and duration time in the observation group were better than those in the control group (both<0.05). The contents of SP in the two groups decreased after treatment (both<0.01), and it was better in the observation group (<0.05). The total effective rate of the observation group after treatment was 87.9% (29/33), and that of the control group was 85.3% (29/34), which were not statistically significant (>0.05). The cured rate of the observation group was better than that of the control group [66.7% (22/33) vs 58.8% (20/34),<0.05].
CONCLUSIONJoss stick moxibustion combined with pricking and cupping are effective for herpes zoster, which have quicker and good analgesic effects than conventional western medication. Its mechanism may be related to reducing the content of SP more fast and to a larger degree.