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Volume 38 Issue 3
Mar.  2022
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2022  >  38(3): 227-235
Luo JH,Zhan JH,Liao WQ,et al.Investigating the effects of Modified Sijunzi Decoction on the diversity of intestinal microflora of severe scald rabbits based on 16S ribosomal RNA high-throughput sequencing[J].Chin J Burns Wounds,2022,38(3):227-235.DOI: 10.3760/cma.j.cn501120-20200923-00421.
Citation: Luo JH,Zhan JH,Liao WQ,et al.Investigating the effects of Modified Sijunzi Decoction on the diversity of intestinal microflora of severe scald rabbits based on 16S ribosomal RNA high-throughput sequencing[J].Chin J Burns Wounds,2022,38(3):227-235.DOI: 10.3760/cma.j.cn501120-20200923-00421.
shu

Investigating the effects of Modified Sijunzi Decoction on the diversity of intestinal microflora of severe scald rabbits based on 16S ribosomal RNA high-throughput sequencing

doi: 10.3760/cma.j.cn501120-20200923-00421

  • Burn and Wound Repair Center, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China

Funds:

Science and Technology Program of Jiangxi Province of China 20171BBG70061, 2020BBG73020

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  • Corresponding author: Zhan Jianhua, Email: zjhseasky@163.com
  • Received Date: 2020-09-23
    Abstract
  •     Objective   To investigate the effects of Modified Sijunzi Decoction on the diversity of intestinal microflora of in severe scald rabbits based on 16S ribosomal RNA (16S rRNA) high-throughput sequencing.    Methods   The experimental research method was adopted. Ninety Japanese big-ear rabbits regardless gender, aged 6 to 8 months, were randomly divided into normal control group, scald alone group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group, with 18 rabbits in each group. The rabbits in normal control group were free to eat and drink, and the rabbits in scald alone group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group were intragastrically administered normal saline, 0.2 g/mL Modified Sijunzi Decoction, 1.0 g/mL Modified Sijunzi Decoction, and 5.0 g/mL Modified Sijunzi Decoction, respectively for 7 days after sustaining full-thickness scalding of 30% total body surface area. On the 1st, 3rd, and 7th day after grouping, the levels of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-10 in ileal mucosa tissue of rabbits in each group were determined by enzyme-linked immunosorbent assay, and the number of samples in each group at each time point was 6. According to the above experimental results, another 9 rabbits were selected and divided into normal control group, scald alone group and scald+medium-dose group, with 3 rabbits in each group. The grouping and treatment methods of rabbits in each group were the same as before. On the 7th day after grouping, the V3, V4 region of 16S rRNA of ileum mucosa of rabbits in three groups were sequenced by high-throughput sequencing technology. The number of quality bacteria was counted by QIME software. The classifications of phylum, class, order, family and genus of microflora were analyzed by RDP Classifier software. The α diversity (Ace, Chao1, Simpson, and Shannon indexes) and β diversity were analyzed by Illumina MiSeq sequencing technology, and the number of experiment samples in each group was 3. Data were statistically analyzed with analysis for variance of factorial design, SNK test, and Bonferroni correction.    Results   Compared with that in normal control group, the levels of TNF-α of ileal mucosa tissue of rabbits in scald alone group, scald+low-dose group, and scald+high-dose group on the 1st, 3rd, and 7th day after grouping and scald+medium-dose group on the 1st and 3rd day after grouping were all significantly increased (P<0.01), the levels of IL-1β in ileal mucosa tissue of rabbits in scald alone group, scald+low-dose group, scald+medium-dose group and scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly increased (P<0.05 or P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald alone group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly decreased (P<0.01). Compared with that in scald alone group, the levels of TNF-α in ileal mucosa tissue of rabbits in scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 3rd and 7th day after grouping, and scald+medium-dose group on the 1st day after grouping were all significantly decreased (P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 3rd and 7th day after grouping and scald+medium-dose group on the 1st day after grouping were all significantly decreased (P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald+low-dose group on the 7th day after grouping and scald+medium-dose group on the 1st, 3rd, and 7th day after grouping and scald+high-dose group on the 3rd and 7th day after grouping were all significantly increased (P<0.05 or P<0.01). Compared with that in scald+low-dose group, the levels of TNF-α in ileal mucosa tissue of rabbits in medium-dose scald alone group on the 1st, 3rd, and 7th day after grouping and in high-dose scald alone group on the 3rd and 7th day after grouping were significantly decreased (P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in medium-dose scald alone group on the 1st, 3rd, and 7th day after grouping and in high-dose scald alone group on the 3rd and 7th day after grouping were all significantly decreased (P<0.05 or P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald+medium-dose group on the 1st, 3rd, and 7th day after grouping and in scald+high-dose group on the 7th day after grouping were all significantly increased (P<0.05 or P<0.01). Compared with that in scald medium-dose group, the levels of TNF-α in ileal mucosa tissue of rabbits in scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly increased (P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly decreased (P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in scald+high-dose group on the 7th day after grouping was significantly decreased (P<0.01). Compared with that on the 1st day after grouping, the levels of TNF-α in ileal mucosa tissue of rabbits in scald alone group on the 3rd and 7th day after grouping and in normal control group on the 3rd day after grouping were all significantly increased (P<0.05 or P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in scald alone group both on the 3rd and 7th day after grouping were significantly increased (P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in both scald+low-dose group and scald+high-dose group on the 7th day after grouping and scald+medium-dose group both on the 3rd and 7th day after grouping were significantly increased (P<0.05 or P<0.01), and the levels of TNF-α in ileal mucosa tissue of rabbits in scald+high-dose group on the 3rd and 7th day after grouping and in scald+medium-dose group on the 7th day after grouping were all significantly decreased (P<0.05 or P<0.01), and the level of IL-1β in ileal mucosa tissue of rabbits in scald+medium-dose group on the 7th day after grouping was significantly decreased (P<0.01), and the level of IL-10 in ileal mucosa tissue of rabbits in scald alone group on the 7th day after grouping was significantly decreased (P<0.01). Compared with that on the 3rd day after grouping, the levels of TNF-α and IL-1β in ileal mucosa tissue of rabbits in scald alone group and the levels of IL-10 in ileal mucosa tissue of rabbits in normal control group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 7th day after grouping were all significantly increased (P<0.05 or P<0.01); and the levels of TNF-α in ileal mucosa tissue of rabbits in scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 7th day after grouping were all significantly decreased (P<0.05), and the levels of IL-1β in ileal mucosa tissue of rabbits both in scald+medium-dose group and scald+high-dose group on the 7th day after grouping were significantly decreased (P<0.05 or P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald alone group on the 7th day after grouping was significantly decreased (P<0.01). On the 7th day after grouping, the high-quality sequences obtained from the microflora in ileum mucosa of rabbits in normal control group, scald alone group, and scald+medium-dose group were 96 023, 107 365, and 95 921, respectively. At the classification level of phylum, class, order, family, and genus of the microflora in ileum mucosa of rabbits in three groups were all Bacteroidetes and Firmicutes, Clostridium and Bacteroidetes, Clostridium and Bacteroidetes, Rumenobacteriaceae and Clostridium and Bacteroideaceae, Clostridium and Bacteroidetes and rumen bacteria mainly, while the percentage of microflora in each group was different. There were no significant differences in Ace, Chao1, Simpson, Shannon indices (P>0.05), and no obvious difference in β diversity of microflora in ileal mucosa tissue of rabbits among three groups.    Conclusions   After severe scalding, the inflammatory response of rabbit ileal mucosa tissue is obvious and increased in a time-dependent manner. Modified Sijunzi Decoction can reduce inflammation with optimal therapeutic concentration of 1.0 g/mL. The technology of high-throughput sequencing can reflect the structural composition of the intestinal microflora accurately. The ileal microflora of the severe scald rabbit can be regulated by the administration of Modified Sijunzi Decoction.

     

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    Created with Highcharts 5.0.7Chart context menuAccess Area Distribution其他: 2.0 %其他: 2.0 %Canton: 0.4 %Canton: 0.4 %China: 11.1 %China: 11.1 %Norman: 0.1 %Norman: 0.1 %Seattle: 0.1 %Seattle: 0.1 %[]: 0.1 %[]: 0.1 %三明: 0.6 %三明: 0.6 %三门峡: 0.2 %三门峡: 0.2 %上海: 0.5 %上海: 0.5 %上饶: 0.1 %上饶: 0.1 %东莞: 0.2 %东莞: 0.2 %丽水: 1.6 %丽水: 1.6 %乐山: 0.1 %乐山: 0.1 %九江: 0.1 %九江: 0.1 %佛山: 0.1 %佛山: 0.1 %保定: 0.1 %保定: 0.1 %信阳: 0.1 %信阳: 0.1 %六安: 0.4 %六安: 0.4 %包头: 0.2 %包头: 0.2 %北京: 0.9 %北京: 0.9 %北方邦: 0.1 %北方邦: 0.1 %南京: 0.8 %南京: 0.8 %南宁: 0.1 %南宁: 0.1 %南平: 1.0 %南平: 1.0 %南昌: 1.3 %南昌: 1.3 %南通: 0.2 %南通: 0.2 %厦门: 1.3 %厦门: 1.3 %台州: 1.4 %台州: 1.4 %合肥: 0.1 %合肥: 0.1 %吉安: 0.1 %吉安: 0.1 %吉林: 0.2 %吉林: 0.2 %呼伦贝尔: 0.1 %呼伦贝尔: 0.1 %咸阳: 1.8 %咸阳: 1.8 %唐山: 0.1 %唐山: 0.1 %嘉兴: 1.2 %嘉兴: 1.2 %大连: 1.4 %大连: 1.4 %宁德: 0.8 %宁德: 0.8 %宁波: 0.9 %宁波: 0.9 %安庆: 0.1 %安庆: 0.1 %宜春: 0.2 %宜春: 0.2 %宣城: 0.1 %宣城: 0.1 %宿迁: 2.0 %宿迁: 2.0 %常德: 0.7 %常德: 0.7 %平顶山: 0.1 %平顶山: 0.1 %广元: 0.2 %广元: 0.2 %广州: 1.2 %广州: 1.2 %廊坊: 0.1 %廊坊: 0.1 %张家口: 0.4 %张家口: 0.4 %徐州: 1.2 %徐州: 1.2 %德州: 0.2 %德州: 0.2 %德阳: 0.1 %德阳: 0.1 %怀化: 0.1 %怀化: 0.1 %成都: 0.1 %成都: 0.1 %扬州: 0.8 %扬州: 0.8 %抚州: 0.5 %抚州: 0.5 %拉贾斯坦邦: 0.2 %拉贾斯坦邦: 0.2 %无锡: 0.7 %无锡: 0.7 %日喀则: 0.1 %日喀则: 0.1 %日照: 0.2 %日照: 0.2 %昆明: 0.4 %昆明: 0.4 %朝阳: 0.6 %朝阳: 0.6 %杭州: 1.6 %杭州: 1.6 %枣庄: 0.1 %枣庄: 0.1 %榆林: 1.2 %榆林: 1.2 %武威: 0.1 %武威: 0.1 %武汉: 0.6 %武汉: 0.6 %汕头: 0.7 %汕头: 0.7 %池州: 0.7 %池州: 0.7 %沈阳: 1.3 %沈阳: 1.3 %泉州: 0.8 %泉州: 0.8 %泰州: 0.4 %泰州: 0.4 %泸州: 0.2 %泸州: 0.2 %济南: 0.4 %济南: 0.4 %海东: 0.1 %海东: 0.1 %海得拉巴: 0.2 %海得拉巴: 0.2 %海西: 0.1 %海西: 0.1 %淮北: 0.3 %淮北: 0.3 %淮南: 0.1 %淮南: 0.1 %淮安: 0.5 %淮安: 0.5 %温州: 0.4 %温州: 0.4 %渭南: 0.9 %渭南: 0.9 %湖州: 0.8 %湖州: 0.8 %湘潭: 0.1 %湘潭: 0.1 %湘西: 0.6 %湘西: 0.6 %湘西土家族苗族自治州: 0.1 %湘西土家族苗族自治州: 0.1 %滁州: 0.1 %滁州: 0.1 %滨州: 0.1 %滨州: 0.1 %漯河: 0.1 %漯河: 0.1 %漳州: 1.1 %漳州: 1.1 %潍坊: 0.1 %潍坊: 0.1 %濮阳: 0.1 %濮阳: 0.1 %珠海: 0.6 %珠海: 0.6 %白城: 0.1 %白城: 0.1 %盐城: 0.4 %盐城: 0.4 %石家庄: 0.4 %石家庄: 0.4 %福州: 0.8 %福州: 0.8 %秦皇岛: 0.4 %秦皇岛: 0.4 %绍兴: 1.3 %绍兴: 1.3 %绵阳: 0.3 %绵阳: 0.3 %自贡: 0.1 %自贡: 0.1 %舟山: 1.2 %舟山: 1.2 %芒廷维尤: 11.6 %芒廷维尤: 11.6 %芝加哥: 0.1 %芝加哥: 0.1 %苏州: 0.3 %苏州: 0.3 %茂名: 0.2 %茂名: 0.2 %荆州: 0.2 %荆州: 0.2 %荆门: 1.1 %荆门: 1.1 %莆田: 1.1 %莆田: 1.1 %萍乡: 0.1 %萍乡: 0.1 %营口: 0.7 %营口: 0.7 %葫芦岛: 0.8 %葫芦岛: 0.8 %蒙彼利埃: 0.1 %蒙彼利埃: 0.1 %衡水: 0.1 %衡水: 0.1 %衢州: 0.2 %衢州: 0.2 %襄阳: 0.1 %襄阳: 0.1 %西宁: 2.5 %西宁: 2.5 %西安: 0.8 %西安: 0.8 %许昌: 0.1 %许昌: 0.1 %辽阳: 0.6 %辽阳: 0.6 %运城: 0.1 %运城: 0.1 %连云港: 0.3 %连云港: 0.3 %通化: 0.1 %通化: 0.1 %遵义: 0.6 %遵义: 0.6 %邯郸: 0.1 %邯郸: 0.1 %邵阳: 0.2 %邵阳: 0.2 %郑州: 0.3 %郑州: 0.3 %郴州: 1.5 %郴州: 1.5 %重庆: 5.2 %重庆: 5.2 %金华: 1.0 %金华: 1.0 %铁岭: 1.5 %铁岭: 1.5 %铜陵: 0.4 %铜陵: 0.4 %锦州: 0.5 %锦州: 0.5 %长春: 0.1 %长春: 0.1 %长沙: 0.1 %长沙: 0.1 %随州: 0.5 %随州: 0.5 %雅安: 0.1 %雅安: 0.1 %青岛: 0.1 %青岛: 0.1 %鞍山: 4.2 %鞍山: 4.2 %韶关: 0.1 %韶关: 0.1 %香港: 0.1 %香港: 0.1 %马鞍山: 0.1 %马鞍山: 0.1 %鹰潭: 0.1 %鹰潭: 0.1 %黄冈: 0.9 %黄冈: 0.9 %黄南: 0.2 %黄南: 0.2 %黄山: 0.1 %黄山: 0.1 %黄石: 0.2 %黄石: 0.2 %其他CantonChinaNormanSeattle[]三明三门峡上海上饶东莞丽水乐山九江佛山保定信阳六安包头北京北方邦南京南宁南平南昌南通厦门台州合肥吉安吉林呼伦贝尔咸阳唐山嘉兴大连宁德宁波安庆宜春宣城宿迁常德平顶山广元广州廊坊张家口徐州德州德阳怀化成都扬州抚州拉贾斯坦邦无锡日喀则日照昆明朝阳杭州枣庄榆林武威武汉汕头池州沈阳泉州泰州泸州济南海东海得拉巴海西淮北淮南淮安温州渭南湖州湘潭湘西湘西土家族苗族自治州滁州滨州漯河漳州潍坊濮阳珠海白城盐城石家庄福州秦皇岛绍兴绵阳自贡舟山芒廷维尤芝加哥苏州茂名荆州荆门莆田萍乡营口葫芦岛蒙彼利埃衡水衢州襄阳西宁西安许昌辽阳运城连云港通化遵义邯郸邵阳郑州郴州重庆金华铁岭铜陵锦州长春长沙随州雅安青岛鞍山韶关香港马鞍山鹰潭黄冈黄南黄山黄石

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