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| Effect of Chinese Herbal Compound Extract on Inflammation of Intestinal Epithelial Cells and Bacterial Diarrhea in Dogs (Canis lupus familiaris) |
| LIU Hai-Xia1, LIU Gen-Xin1,*, TIAN Qi-Hui1, ZHAO Tian2, ZHANG Jie-Hui1, ZHU Yan-Hua1, LIU Di-Qin1, GUO Zhi-Ming1 |
1 Gansu Polytechnic College of Animal Husbandry & Engineering, Wuwei 733006, China;
2 College of Veterinary Medicine, Gansu Agricultural University |
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Abstract Canine bacterial diarrhea is a common diarrhea disease caused by pathogenic bacteria in dogs (Canis lupus familiaris). Antibiotics are often used in clinical treatment, which also increases the incidence of drug-resistant bacteria. Qinlian Qinpi Decoction has good effect in the treatment of bacterial diarrhea in young children. In order to explore the therapeutic mechanism of Qinlian Qinpi Decoction in canine bacterial diarrhea, this study used semi-bionic enzyme extraction method to extract the effective components of Qinlian Qinpi Decoction, and the antibacterial effect of the extract was detected by punching method, the cytotoxicity was detected by Cell Counting Kit-8 (CCK-8), and dog intestinal epithelial cells were infected with Escherichia coli. The levels of Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB), interleukin 1β (IL1β), IL6, proliferating cell nuclear antigen (PCNA), B cell lymphoma-2 (Bcl2) and Bcl2-associated X (Bax) gene and protein were detected by qRT-PCR, Western blot and ELISA, and the cell apoptosis was detected by TUNEL method. Finally, the clinical therapeutic effect of the extract on canine bacterial diarrhea was determined by observation method. The results showed that the extract effectively inhibited the growth of E. coli in a certain concentration range and time without cytotoxicity. In canine intestinal epithelial cells infected with large intestine, the extract effectively inhibited the transcription levels and protein expression of TLR4 and NF-κB, and inhibited the transcription levels and cell secretion levels of IL-1β and IL-6. At the same time, the extract had obvious effects in anti-apoptosis and promoting cell proliferation. The results of in vivo experiments showed that the extract was effective for canine bacterial diarrhea, and when combined with ceftriaxone sodium, the treatment time could be shortened on the basis of effective treatment. The above results showed that the extract of Qinlian Qinpi Decoction could be applied to the treatment of bacterial diarrhea in dogs and helped to reduce the use of antibiotics in clinical treatment. This study provides basic data for the application of Qinlian Qinpi Decoction in the clinical treatment of bacterial diarrhea in dogs.
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Received: 16 December 2022
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Corresponding Authors:
*gsxmxgclgx@163.com
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[1] 彪雅宁, 王瑞龙, 储心乔, 等. 2020. 车前子对腹泻模型大鼠小肠组织水通道蛋白4表达的影响[J]. 国际中医中药杂志, 42(08): 758-762.
(Biao Y N, Wang R L, Chu X Q, et al.2020. Effect of plantaginis semen on expression of aquaporin 4 in small intestine of diarrhea model rats[J]. International Journal of Traditional Chinese Medicine, 42(08): 758-762.)
[2] 戴柳江, 张卉, 李梦怡, 等. 2014. 半仿生-酶法提取中药不同成分的比较研究[J]. 西北药学杂志, 29(04): 331-335.
(Dai L J, Zhang H, Li M Y, et al.2014.Comparative study of three different herbs extracted by semi-bionic enzyme extraction[J]. Northwest Pharmaceutical Journal, 29(04): 331-335.)
[3] 顾楠. 2020. 芩连秦皮汤治疗儿童(细菌性)腹泻的临床研究 [D]. 硕士学位论文, 长春中医药大学, 导师: 王永吉, pp. 2-24.
(Gu N.2020. The clinical observation of Qinlianqinpi Decoction in the controltreatment of diarrhea in children (bacterial)[D]. Thesis for M.S., Changchun University of Chinese Medicine, Supervisor: Wang Y J, pp. 2-24.)
[4] 韩阳春. 2022. 白芍总苷对ETEC致腹泻仔猪肠道基因转录组学变化及炎性损伤的调控作用[D]. 硕士学位论文, 西南大学,导师: 彭远义, 王自力, pp. 43-53.
(Han Y C.2022. Regulation of total glucosides of paeony on intestinal transcriptome changes and inflammatory injury in diarrhea piglets induced by ETEC[D]. Thesis for M.S., Southwest University, Supervisor: Pen Y Y, Wang Z L, pp.43-53.)
[5] 黄春丽, 冯光维, 余跃生, 等. 2022. 泽泻对MSG大鼠IL-6、TNF-α含量和基因表达的影响[J]. 中国民族民间医药, 31(03): 17-20.
(Huang C L, Feng G W, Yu Y S, et al.2022. Effect of Alismatis rhizoma on IL-6,TNF-α content and gene expression in MSG rats[J]. Chinese Joumal of Ethnomedicine and Ethnopharmacy, 31(03): 17-20)
[6] 金振华, 高利. 2017. 犬腹泻的常见类型、临床症状、诊断方法及综合治疗[J]. 现代畜牧科技, (04): 110.
(Jin Z H, Gao L. 2017. Common types, clinical symptoms, diagnosis and comprehensive treatment of diarrhea in dogs[J]. Technical Advisor for Animal Husbandry, (04): 110)
[7] 李晓燕. 2015. 犬腹泻的中西医结合治疗法[J]. 中国畜牧业, (09): 84.
(Li X Y. 2015. Integrative therapy for diarrhea in dogs[J]. China Animal Industry, (09): 84.)
[8] 刘瑞雪. 2022. 刺五加总黄酮提取纯化及其对氧化应激诱导凋亡的研究[D]. 硕士学位论文, 聊城大学, 导师: 苏建青, 褚秀玲, pp. 27-39.
(Liu R X.2022. Extraction and purification of total flavonoids from Acanthopanax senticosus and their effects on oxidative stress-induced apoptotic[D]. Thesis for M.S., Liaocheng University, Supervisor: Su J Q, Chu X L, pp. 27-39.)
[9] 区耿华, 让一峰, 黄卓权, 等. 2021. 佛手提取物的化学成分和生物活性研究进展[J]. 食品科技, 46(09): 169-174.
(Ou G H, Rang Y F, Huang Z Q, et al.2021. Research progress on chemical constituents and biological activities of Citrus medica L. var. Sarcodactylis swingle extract[J]. Food Science and Technology, 46(09): 169-174.)
[10] 潘嘉臻. 2021. 杭州地区犬、猫肠道菌群分离鉴定及其耐药分析[D]. 硕士学位论文, 浙江大学, 导师: 王华南, pp. 2-10.
(Pan J Z.2021. Characterization and antimicrobial resistance analysis of important intestinal bacteria recovered from canine and feline in Hangzhou[D]. Thesis for M.S., Zhejiang University, Supervisor: Wang H N, pp. 2-10.)
[11] 王淑玲. 2009. 半夏白术天麻汤用半仿生—生物酶法提取的系统研究[D]. 博士学位论文, 山东中医药大学, 导师: 孙秀梅, pp. 07-11.
(Wang S L.2009. Studies on Banxia Baizhu Tianma Decoction by semi-bionic enzyme extraction mode[D]. Thesis for Ph.D., Shandong University of Traditional Chinese Medicine, Supervisor: Sun Y M, pp. 7-11.)
[12] 王涛, 王玉金, 朱贵霞. 2013. 宠物犬细菌性腹泻的诊断和治疗[J]. 中国动物保健, 15(09): 73.
(Wang T, Wang Y J, Zhu G X.2013. Diagnosis and treatment of bacterial diarrhea in pet dogs[J]. China Animal Health, 15(09): 73.)
[13] 魏媛媛, 张艳楠, 樊艺萌, 等. 2022. 中兽药创新研发与实验动物模型应用[J]. 中国比较医学杂志, 32(1): 97-103.
(Wei Y Y, Zhang Y N, Fan Y M, et al.2022. Innovative research and development of Chinese veterinary drugs and application of experimental animal models[J]. Chinese Journal of Comparative Medicine, 32(1): 97-103.)
[14] 许剑琴, 王自力, 麻武仁, 等. 2013. 犬猫腹泻辨证施治[J]. 中兽医学杂志, (02): 35-38.
(Xu J Q, Wang Z L, Ma W R, et al. 2013. Dialectical treatment of dog and cat diarrhea[J]. Chinese Journal of Traditional Veterinary Science, (02): 35-38.)
[15] 夏永高, 李维功, 刘桂清, 等. 2004. 中西医结合治疗犬腹泻[J]. 中国兽医科技, (06): 69-70.
(Xia Y G, Li W G, Liu G Q, et al. 2004. Treatment of canine diarrhea by integrative medicine[J]. Chinese Journal of Veterinary Science and Technology, (06): 69-70.)
[16] 仙靓, 郭增军, 覃源芮, 等. 2018. 半仿生-酶法提取虎杖中大黄素的工艺研究[J]. 西北药学杂志, 33(02): 158-160.
(Xian L, Guo Z J, Qin Y R, et al.2018. Extraction of emodin from Reynoutria japonica Houtt. by semi-bionic enzyme method[J]. Northwest Pharmaceutical Journal, 33(02): 158-60.)
[17] 张传美, 徐尘枫, 邹玲, 等. 2019. 腹泻犬肠道分离菌的鉴定及耐药性分析[J]. 中国兽医杂志, 55(10): 82-85, 90.
(Zhang C M, Xu C F, Zou L, et al.2019. Identification and detection of drug resistance on pathogenic bacteria from canine diarrhea[J]. Chinese Journal of Veterinary Medicine, 55(10): 82-85, 90.)
[18] 张红阳, 曹世杰, 邱峰, 等. 2019. 中药活性成分抑制抗凋亡蛋白抗肿瘤研究进展[J]. 天津中医药, 36(08): 827-832.
(Zhang H Y, Cao S J, Qiu F, et al.2019. Research progress in inhibiting anti-apoptotic proteins of active components of traditional Chinese medicine for anti-tumor[J]. Tianjin Journal of Traditional Chinese Medicine, 36(08): 827-832.)
[19] 张华, 王伟, 赵学修. 2006. 中西医结合治疗犬腹泻症[J]. 现代农业科技, (03): 79-80.
(Zhang H, Wang W, Zhao X X. 2006. Treatment of canine diarrhea by integrative medicine[J]. Modern Agricultural Science and Technology, (03): 79-80.)
[20] 张越. 2022. 砂仁中黄酮类化合物的提取及其生物活性研究[D]. 硕士学位论文, 吉林大学, 导师: 滕利荣, pp. 36-56.
(Zhang Y.2022. Study on extraction and biologicalactivity of flavonoid from Amomum villosum[D]. Thesis for M.S., Jilin University, Supervisor: Teng L R, pp. 36-56.)
[21] 赵宏, 陈晨, 赵岩, 等. 2021. 车前子多糖对膜性肾病大鼠肾损伤和肠道菌群的影响[J]. 中国实验方剂学杂志, 27(22): 92-99.
(Zhao H, Chen C, Zhao Y, et al.2021. Effect of polysaccharides from plantaginis semen on renal injury and gut microbiota in rats with membranous nephropathy[J]. Chinese Journal of Experimental Traditional Medical Formulae, 27(22): 92-99.)
[22] 郑晨曦. 2022. 葛根芩连汤改善溃疡性结肠炎小鼠肠黏膜屏障的作用机制研究[D]. 硕士学位论文, 长春中医药大学, 导师: 孙聪, pp.16-46.
(Zheng X C.2022. Mechanism of Gegen Qinlian Decoction improvingintestinal mucosal barrier in mice with ulcerative colitis[D]. Thesis for M.S., Changchun University of Chinese Medicine, Supervisor: Sun C, pp. 16-46.)
[23] Hall E J.2011. Antibiotic-responsive diarrhea in small animals[J]. The Veterinary Clinics of North America: Small Animal Practice, 41(2): 273-286.
[24] Huang Z, Pan Z, Yang R, et al.2020. The canine gastrointestinal microbiota: Early studies and research frontiers[J]. Gut Microbes, 11(4): 635-654.
[25] Huang Y, Cai T, Xia X, et al.2016. Research advances in the intervention of inflammation and cancer by active ingredients of traditional Chinese medicine[J]. Journal of Pharmacy and Pharmaceutical Sciences, 19(1): 114-126.
[26] Kalenyak K, Isaian A, Heilmann R M, et al.2018. Comparison of the intestinal mucosal microbiota in dogs diagnosed with idiopathic inflammatory bowel disease and dogs with food-responsive diarrhea before and after treatment[J]. FEMS microbiology ecology, 94(2): 1-11.
[27] Marks S L, Kather E J.2003. Bacterial-associated diarrhea in the dog: A critical appraisal[J]. The Veterinary Clinics of North America. Small Animal Practice, 33(5): 1029-1060.
[28] Marks S L, Rankin S C, Byrne B A, et al.2011. Enteropathogenic bacteria in dogs and cats: Diagnosis, epidemiology, treatment, and control[J]. Journal of Veterinary Internal Medicine, 25(6): 1195-1208.
[29] Minamoto Y, Dhanani N, Markel M E, et al.2014. Prevalence of Clostridium perfringens, Clostridium perfringens enterotoxin and dysbiosis in fecal samples of dogs with diarrhea[J]. Veterinary Microbiology, 174: 463-473.
[30] Schirmer M, Garner A, Vlamakis H, et al.2019. Microbial genes and pathways in inflammatory bowel disease[J]. Nature Reviews Microbiology, 17(8): 497-511.
[31] Werner M, Unterer S.2021. Use of antimicrobials in acute canine diarrhea - overview of potential risks, indications and alternatives[J]. Tierarztliche Praxis, 49(2): 110-120. |
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