CN113350384A - Application of hydrangea paniculata extract in treating chronic nephropathy by regulating intestinal flora - Google Patents
Application of hydrangea paniculata extract in treating chronic nephropathy by regulating intestinal flora Download PDFInfo
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- CN113350384A CN113350384A CN202010146995.8A CN202010146995A CN113350384A CN 113350384 A CN113350384 A CN 113350384A CN 202010146995 A CN202010146995 A CN 202010146995A CN 113350384 A CN113350384 A CN 113350384A
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- lactobacillus
- hydrangea paniculata
- clostridium
- paniculata extract
- ruminococcaceae
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Abstract
The invention discloses an application of hydrangea paniculata extract in preparing a medicine for preventing or treating chronic nephropathy, wherein the hydrangea paniculata extract is extracted by reflux extraction, macroporous resin ion exchange and reduced pressure distillation, and the hydrangea paniculata extract is used for preventing and treating chronic nephropathy by adjusting intestinal flora. The invention discloses a new mechanism for relieving chronic nephropathy by hydrangea paniculata extract; the hydrangea paniculata extract is applied to the preparation of medicines for regulating immune nephropathy and intestinal flora disorder caused by the immune nephropathy. The hydrangea paniculata extract can effectively reduce urine albumin and blood creatinine of an organism, effectively improve renal function, regulate and control the levels of total cholesterol, high density lipoprotein, low density lipoprotein and triglyceride in serum, and regulate lipid metabolism.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to application of a hydrangea paniculata extract in treating chronic nephropathy by adjusting intestinal flora.
Background
Kidney disease is a major cause of death worldwide, with about 10% of the world's population suffering from chronic kidney disease and increasingly being recognized as a global health problem. Current treatment of early Chronic Kidney Disease (CKD) focuses mainly on blood pressure control, reduction of protein and salt intake, prevention of acute kidney injury and glycemic control, and CKD has no cure or preventive strategy, and timely treatment is extremely challenging due to lack of symptoms at early stages of the disease. In addition, there is a lack of effective treatment for End Stage Renal Disease (ESRD) in addition to dialysis and kidney transplantation. Thus, there is a need for a concept that changes the paradigm and innovative methods to detect, manage, control and ultimately cure these diseases. At present, the drugs widely used for treating chronic kidney diseases in clinic mainly comprise chemotherapeutic drugs and various hormones or immunosuppressive agents, but the treatment effect is inconsistent, the side effect is serious and the like, so that the treatment of the chronic kidney diseases has great limitation.
Intestinal flora
The human body carries a large amount of bacteria, about 1014These bacteria have structural and histological functions and play an important role in regulating host health and homeostasis. As a "second human genome," the microbiome encodes 330 ten thousand unique genes, 150-fold more than the human genome. The gut microbiome is thus able to perform various physiological metabolic functions that humans cannot perform. In normal human intestinal tracts, the number and the types of intestinal bacteria are large, the intestinal bacteria have multiple functions of substance metabolism, biological barrier, immune regulation, host defense and the like, and different levels of evidence of various animal models and human researches support the action of intestinal microbiota in human health. The intestinal flora is an important research direction in the fields of metabolism and diseases, and is not only basic research but also closely related to the health of our lives. Composition of intestinal microbiotaAnd functional imbalance is closely related to the occurrence and development processes of various diseases, such as inflammatory bowel disease, diabetes, hypertension, obesity, tumor, cardiovascular diseases, chronic kidney diseases and the like. Improving conditions of gut dysbacteriosis may be a potential strategy for the prevention and management of these diseases.
Chronic nephropathy is associated with intestinal flora
There is increasing evidence that gut microbiota plays an important role in the development of CKD. The intestinal flora is thought to be involved in the development of chronic kidney disease. The currently accepted core view of the theory of the 'intestinal-renal axis' is that in the process of disease development, a chronic kidney disease patient has intestinal micro-ecosystem disorder to cause intestinal bacteria imbalance, on one hand, enterogenic uremic toxins accumulate and cannot be timely removed by an injured kidney, and renal function is reduced, and on the other hand, the imbalanced intestinal flora can damage the intestinal epithelial barrier function, so that the enterogenic uremic toxins and conditional pathogenic bacteria are displaced to enter blood circulation, an intestinal mucosal immune system is activated, systemic micro-inflammatory reaction is induced, and kidney injury is aggravated. The gut-kidney axis can be subdivided into metabolic-dependent pathways and immune pathways. The metabolism-dependent pathway is mediated primarily by metabolites produced by the gut microbiota, which have the ability to regulate the physiological functions of the host. In the immune pathway, components of the immune system (e.g., lymphocytes, monocytes, and cytokines) play a crucial role in communication between the gut and the kidney. The interaction between metabolic dependence and the immune pathway also plays an important role in maintaining gut-renal axis balance.
Hydrangea paniculata, chronic nephrosis and intestinal flora
The hydrangea paniculata extract is used as five types of new traditional Chinese medicines and is currently in the preclinical research stage. According to records of Chinese materia medica resource Zhi Yao, the whole hydrangea paniculata plant can clear heat and resist malaria, check malaria and defervesce, eliminate food retention and regulate the middle warmer.
According to the previous experimental results, the hydrangea paniculata extract HP has a good curative effect on chronic kidney disease models such as immune kidney disease and the like, the progress of the disease can be well improved, the specific target point is unknown, the inventor guesses that the hydrangea paniculata extract can improve the kidney function by adjusting intestinal flora according to the disease state of a rat and the preliminary sequencing result and the gradual maturity of the theory of the intestinal renal axis, the method becomes a new method for treating the chronic kidney disease, and the mechanism of treating the chronic kidney disease by the hydrangea paniculata extract HP is explained from another angle.
Disclosure of Invention
The invention aims to solve the technical problems, provides a group of characteristic bacteria for regulating chronic kidney diseases, and provides a novel method for treating chronic kidney diseases, namely, hydrangea paniculata extract is used for effectively regulating intestinal flora of patients with immune nephropathy, normalizing the intestinal flora, reducing urine albumin of organisms and achieving the purpose of effectively treating the nephrotic syndrome of the immune nephropathy.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention firstly provides an application of hydrangea paniculata extract in preparing a medicament for preventing or treating chronic kidney diseases, wherein the hydrangea paniculata extract is extracted by reflux extraction, macroporous resin ion exchange and reduced pressure distillation, and the hydrangea paniculata extract is used for preventing or treating chronic kidney diseases by regulating intestinal flora.
Preferably, 65% of the weight of the hydrangea paniculata extract is coumarin glycoside compounds.
Preferably, the coumarin glycoside compounds mainly comprise skimmin, hydrangea glycoside, scopolamine, 8-methoxycoumarin-7-O-beta-D-glucoside and umbelliferone-7-O-beta-D-glucosyl- (1 → 6) -O-beta-D-glucoside.
Preferably, the method for extracting the coumarins glycosides compound extracted from hydrangea paniculata comprises the following steps:
(1) pulverizing hydrangea paniculata branches, adding 10 times of water, reflux-extracting for 2 times, each time for 3 hr, filtering the extractive solution while it is hot, and mixing filtrates;
(2) passing the filtrate through macroporous adsorbent resin column (HPD100), eluting with 3 times of water of macromolecular polymer volume to remove impurities, and discarding water solution; desorbing coumarin glycoside with 5 times of 17% ethanol, recovering ethanol from eluate under reduced pressure, concentrating to obtain extract, and drying to obtain hydrangea paniculata extract;
(3) dissolving the extract with water, passing the solution through a macromolecular polymer column HP2MGL, eluting with 3 times of water to remove impurities; desorbing coumarin glycoside with 3 times of macromolecular polymer volume of 25% ethanol, evaporating eluate under reduced pressure, and vacuum drying to obtain light brown powder containing various total coumarin glycosides (content (weight) of hydrangea coumarin glycosides is 65%).
Preferably, the hydrangea paniculata extract is used for treating chronic kidney disease by regulating intestinal flora, normalizing the intestinal flora, reducing urinary albumin and serum creatinine, effectively improving renal function, regulating and controlling the levels of total cholesterol, high density lipoprotein, low density lipoprotein and triglyceride in serum, regulating lipid metabolism, relieving glomerular fibrosis degree and reducing lymphocyte infiltration in renal interstitium.
Preferably, the hydrangea paniculata extract can change the intestinal flora structure, so that the abundances of firmicutes and bacteroidetes are reduced, and the ratio of firmicutes to bacteroidetes is reduced.
Preferably, the hydrangea paniculata extract can reverse the imbalance of intestinal flora.
Preferably, the enteric bacteria include Bifidobacterium (Bifidobacterium), Butyrivibrio (Butyrivibrio butyrate), Shigella (Shigella), Enterobacter (Enterobacter), Lactobacillus (Lactobacillus), Prevotella _9 (Proteus _9), Lachnospiraceae _ NK4A136_ group (Proteus _ NK4A136 genus), Prevotella _1 (Proteus 1), Bacteroides (Bacteroides), Allopreviella (Prevotella), Prevotella _ UCG _001 (Prevotella), Ruminococcaceae _ UCG _001 (Proteus _ UCG001 genus), Ruminococcaceae _ UCG _005 (Ruminococcaceae _ UCG005 genus), Turcibacter (Lactobacillus), Allocicola (Allobacter (Mycobacterium), Ruminococcaceae _ UCG _001), Ruminococcaceae _ UC (Lactobacillus), Rutaceae _ UCG _ 3 (Lactobacillus), Rutaceae _ Ctenococcus (Lactobacillus), Ruminococcobacillus (Lactobacillus), Ruminococcus (Lactobacillus) (Streptococcus 3), Streptococcus 3 (Streptococcus) Ruminoiclastidium _6 (Clostridium), Roseburia (Roseburia), Parasterella (ParaSasa), Ruminoiclastidium _9 (Clostridium), Christenseella _ R _7_ group (Klistenstraceae R7), Oscilobacter (Oscillatoria), Anaerortrum (Anaerobiospirillum), Lachnospiraceae _ UCG _005 (LacG 005), Clostridium _ Sensu _ stricoto _1 (Clostridium), Eubacterium _ coprostagenes _ group (Youngobacter).
Further, the invention provides an application of the hydrangea paniculata extract in a medicine for regulating intestinal flora.
Preferably, the hydrangea paniculata extract is extracted by reflux extraction, macroporous resin ion exchange and reduced pressure distillation.
The specific method comprises the following steps:
(1) pulverizing hydrangea paniculata branches, adding 10 times of water, reflux-extracting for 2 times, each time for 3 hr, filtering the extractive solution while it is hot, and mixing filtrates;
(2) passing the filtrate through macroporous adsorbent resin column (HPD100), eluting with 3 times of water of macromolecular polymer volume to remove impurities, and discarding water solution; desorbing coumarin glycoside with 5 times of 17% ethanol, recovering ethanol from eluate under reduced pressure, concentrating to obtain extract, and drying to obtain hydrangea paniculata extract;
(3) dissolving the extract with water, passing the solution through a macromolecular polymer column HP2MGL, eluting with 3 times of water to remove impurities; desorbing coumarin glycoside with 3 times of macromolecular polymer volume of 25% ethanol, evaporating eluate under reduced pressure, and vacuum drying to obtain light brown powder containing various total coumarin glycosides (with hydrangea paniculata coumarin glycoside content of 65%).
Preferably, 65% of the weight of the hydrangea paniculata extract is coumarin glycoside compounds.
Preferably, the coumarin glycoside compounds mainly comprise skimmin, hydrangea glycoside, scopolamine, 8-methoxycoumarin-7-O-beta-D-glucoside and umbelliferone-7-O-beta-D-glucosyl- (1 → 6) -O-beta-D-glucoside.
Preferably, the hydrangea paniculata extract can improve the intestinal flora structure, reverse the intestinal flora imbalance and adjust the diversity of intestinal microorganisms.
Preferably, the hydrangea paniculata extract can improve the intestinal flora structure, so that the abundances of firmicutes and bacteroidetes are reduced, and the ratio of firmicutes to bacteroidetes is reduced.
Preferably, the hydrangea paniculata extract is capable of reversing imbalance of intestinal flora including Bifidobacterium (Bifidobacterium), Butyrivibrio (Butyrivibrio butyricum), Shigella (Shigella flexneri), Enterobacter (Enterobacter), Lactobacillus (Lactobacillus), Prevotella _9 (Proteus _9), Lachnospiraceae _ NK4A136_ group (Lachnospiraceae _ NK4A 136), Prevotella _1 (Proteus 1), Bacteroides (Bacteroides), Allopretella (Prevotella), Prevotella _ UCG _001 (Proteus _ UCG 001), Ruminococcaceae _ UCG _005 (Ruminococcaceae _ UCG 005), Turcinica (Lactobacillus), Rumococcus _ UCG (Lactobacillus), Ruminococci _ 014), Ruminococcaceae _ UCG (Rumococcus), Ruminococcaceae _ UCG (Streptococcus), Ruminococci (Lactobacillus), Ruminococcaceae _1 (Streptococcus) Prevotellaceae _ NK3B31_ group (Prevoteriaceae _ NK3B 31), Lachnospiraceae _ UCG _001 (UCG 001 of the family Lachnospiraceae), Ruminostrobilum _6 (Clostridium), Roseburia (Ropelelia), Paracutella (ParaSaturella), Ruminostrobilum _9 (Clostridium), Christenseella _ R _7_ group (R7 of the family Clitentaceae), Oscilobacter (Oscillatoria), Anaerotruncus (Anaerobiospirillum), Lachnospiraceae _ UCG _005 (UCG 005 of the family Lachnospiraceae), Clostridium _ sensu _ Stricto _1 (Clostridium), Eubacterium ] coriobacter _ group (Clostridium).
Preferably, the hydrangea paniculata extract can normalize flora, protect intestinal mucosa, reduce enterogenic endotoxin entering blood circulation, and effectively relieve chronic nephropathy.
Further, the invention also provides application of the biomarker of chronic kidney disease in preparing a kit or reagent for detecting chronic kidney disease, wherein the biomarker of chronic kidney disease at least comprises any one or more groups of the following microbial flora:
prevotellaceae _ UCG _001 (Prevoteriaceae _ UCG 001), Rikenella (physical research family), Alisipes (Allistipes), Proteobacteria (Proteobacteria), Turcibacter (Zuricobacteria), Clostridium _ vadinBB60_ group (Clostridium order vadinBB 60), Ruminococcaceae _ UCG _010 (Ruminococcaceae _ UCG _ 010), Clostridiaceae _1 (Clostridium _1), Prevotella _1 (Prevotella 1), Prevotella _9 (Proteobacteria _9), Lachospiraceae (Bucales), Streptococcus (Streptococcus), Allobacterium (Mycoplasma), Fusilenibacillus (Mycobacterium), RuminococcyG 008 (Lactobacillus), Ruminococcaceae _ UCG (Lactobacillus), Lactobacillus paracoccus) (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus) and Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus) and Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus), and Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), and Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccu, Blautia (Blautia), ruminococcus _ UCG _014 (UCG 014 of the family Ruminococcaceae), Desulfovibrio (devulcanium), Helicobacter (Helicobacter), romboutia (rossia), Prevotellaceae _ NK3B31_ group (Prevotellaceae _ NK3B 31), ruminifloride _6 (Clostridium), Roseburia (rocarponia), parasitella (parasternate), ruminifloride _9 (Clostridium), christiaceae _ R _7_ group (klebsiella R7), oscillobacter (oscillaceae), antrotryocus (anaerobacter), Clostridium _ ucruritium _1 (Clostridium).
Preferably, the flora Prevotellaceae _ UCG _001 (Prevoteriaceae _ UCG001 genus), Rikenella (Hodgradae family), Alisipes (Allophyllaceae family), Proteobacteria (Proteobacteria), Turcibacter (Zuricobacter), Clostridiaceae _ vadinBB60_ group (Clostridiaceae vaBB 60 genus), Ruminococcaceae _ UCG _010 (Ruminococcaceae _ UCG _010 genus), Clostridiaceae _1 (Clostridium _1) have a high abundance in HP administration group samples, and Prevotella _1 (Proteobacteria _1), Prevotella _9 (Proteobacteria _9), Lachnospiraceae (Lactuchacteriaceae family), Streptococcus (Streptococcus), Albacteria (Albacteriaceae genus), Fucales (Fusarium 008), Ruminococcaceae _ UCG _ model (Anacardiaceae) have a high abundance in HP administration group samples.
Preferably, the flora Lactobacillus (Lactobacillus), Bifidobacterium (Bifidobacterium), Prevotellaceae _ UCG _001 (Prevotellaceae _ UCG001 genus), Butyrivibrio (vibrio butyrate), Alistipes (Alistipes) are less abundant in the disease model sample; the Escherichia _ Shigella (Shigella), Blautia (Blattella), Allobaculum (Mycoplasma), Enterobacter (Enterobacter) were more abundant in the disease model group samples.
Preferably, the kit or reagent comprises: has the sequence shown in SEQ ID NO: 1 and SEQ ID NO: 2.
Preferably, the kit provides information for diagnosis of chronic kidney disease by isolating DNA in a sample from a subject, performing a PCR reaction on the isolated DNA using the above primers, sequencing, comparing the content of the biomarker of chronic kidney disease with that of a sample from a normal individual, and determining that the content of the biomarker of chronic kidney disease in the sample from the subject is increased or decreased.
Still further, the present invention provides the use of a biomarker of chronic kidney disease comprising at least a group of any one or more of the following microbial flora:
prevotellaceae _ UCG _001 (Prevoteriaceae _ UCG 001), Rikenella (physical research family), Alisipes (Allistipes), Proteobacteria (Proteobacteria), Turcibacter (Zuricobacteria), Clostridium _ vadinBB60_ group (Clostridium order vadinBB 60), Ruminococcaceae _ UCG _010 (Ruminococcaceae _ UCG _ 010), Clostridiaceae _1 (Clostridium _1), Prevotella _1 (Prevotella 1), Prevotella _9 (Proteobacteria _9), Lachospiraceae (Bucales), Streptococcus (Streptococcus), Allobacterium (Mycoplasma), Fusilenibacillus (Mycobacterium), RuminococcyG 008 (Lactobacillus), Ruminococcaceae _ UCG (Lactobacillus), Lactobacillus paracoccus) (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus) and Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus) and Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus), Lactobacillus paracoccus (Lactobacillus paracoccus), Lactobacillus paracoccus (Lactobacillus paracoccus, Blautia (Blautia), ruminococcus _ UCG _014 (UCG 014 of the family Ruminococcaceae), Desulfovibrio (devulcanium), Helicobacter (Helicobacter), romboutia (rossia), Prevotellaceae _ NK3B31_ group (Prevotellaceae _ NK3B 31), ruminifloride _6 (Clostridium), Roseburia (rocarponia), parasitella (parasternate), ruminifloride _9 (Clostridium), christiaceae _ R _7_ group (klebsiella R7), oscillobacter (oscillaceae), antrotryocus (anaerobacter), Clostridium _ ucruritium _1 (Clostridium).
Preferably, the medicament is capable of increasing or decreasing the abundance or amount of the aforementioned biomarkers of chronic kidney disease.
Further, the present invention provides a food, a probiotic, or a medicament for the treatment or prevention of chronic kidney disease, which is capable of increasing or decreasing the abundance or content of the above-mentioned biomarker for chronic kidney disease.
In a preferred embodiment, the food, probiotic or medicament for the treatment or prevention of chronic kidney disease comprises one or more of Prevotellaceae _ UCG _001 (Prevotellaceae _ UCG 001), rikenella (rikennellaceae), Alistipes (cladosporium), tulicibacter (zurich genus), Proteobacteria (Proteobacteria), clostridium _ vatinb 60_ group (Clostridiales vadbb 60), ruminococcus _ UCG _010 (Ruminococcaceae _ UCG _ 010), clostridium _1 (clostridium _1), Lactobacillus (Lactobacillus), Bifidobacterium (Bifidobacterium), butryivibrio (vibrio butyrate).
Further, the invention provides the use of a biomarker of chronic kidney disease as a target for screening a medicament for treating or preventing chronic kidney disease, wherein the biomarker of chronic kidney disease at least comprises any one or more of the following groups of microbial flora:
prevotellaceae _ UCG _001 (Prevoteriaceae _ UCG 001), Rikenella (physical research family), Alisipes (Allistipes), Proteobacteria (Proteobacteria), Turcibacter (Zuricobacteria), Clostridium _ vadinBB60_ group (Clostridium order vadinBB 60), Ruminococcaceae _ UCG _010 (Ruminococcaceae _ UCG _ 008), Clostridium _1 (Clostridium _1), Prevotella _1 (Prevotella 1), Prevotella _9 (Proteobacteria _9), Lachnospirilaceae (Lactobacillaspiraceae), Streptococcus (Streptococcus), Allobacterium (Mycoplasma), Fusilenibacillus (Mycobacterium), RuminococcyG (Corynebacterium 008), Lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus), Lactobacillus (Lactobacillus) Lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) strain, Lactobacillus) and Lactobacillus) Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) strain, Lactobacillus) and Lactobacillus (Lactobacillus), Lactobacillus) strain (Lactobacillus), Lactobacillus (Lactobacillus), Lactobacillus (Lactobacillus) strain (Lactobacillus), Lactobacillus) strain (Lactobacillus), Lactobacillus (Lactobacillus) or Lactobacillus) strain (Lactobacillus) and Lactobacillus) strain (Lactobacillus) and Lactobacillus (Lactobacillus) of the like, Blautia (Blautia), ruminococcus _ UCG _014 (UCG 014 of the family Ruminococcaceae), Desulfovibrio (devulcanium), Helicobacter (Helicobacter), romboutia (rossia), Prevotellaceae _ NK3B31_ group (Prevotellaceae _ NK3B 31), ruminifloride _6 (Clostridium), Roseburia (rocarponia), parasitella (parasternate), ruminifloride _9 (Clostridium), christiaceae _ R _7_ group (klebsiella R7), oscillobacter (oscillaceae), antrotryocus (anaerobacter), Clostridium _ ucruritium _1 (Clostridium).
Advantageous effects
The invention discloses a new mechanism for relieving chronic nephropathy by hydrangea paniculata extract. The effect of HP in treating the immune nephropathy is explored from the angle of regulation of intestinal flora, the intestinal flora of an immune nephropathy model is effectively regulated through the hydrangea paniculata extract, the intestinal flora is normalized, the urinary albumin of the body is reduced, and the aim of effectively treating the nephrotic syndrome of the immune nephropathy is fulfilled.
The hydrangea paniculata extract is applied to the preparation of medicines for regulating immune nephropathy and intestinal flora disorder caused by the immune nephropathy. The hydrangea paniculata extract can effectively reduce urine albumin and blood creatinine of an organism, effectively improve renal function, regulate and control the levels of total cholesterol, high density lipoprotein, low density lipoprotein and triglyceride in serum, and regulate lipid metabolism.
In order to further discover a new mechanism for improving immune nephropathy by HP, the inventor discovers characteristic bacteria related to immune nephropathy and HP intervention through 16s sequencing, and simultaneously proves that HP can remarkably increase the types and the number of flora related to kidney protection, normalize the flora, protect intestinal mucosa, reduce enterogenic endotoxin from entering blood circulation, and effectively relieve the occurrence of chronic nephropathy.
Drawings
FIG. 1HE staining results; n represents Normal, Normal group; m represents Model, Model group; MMF, representing the group given mycophenolate mofetil treatment; HP, representing treatment group given hydrangea paniculata extract.
FIGS. 2A-H are sequencing results analysis. Wherein the number of OTUs shared and unique among groups (fig. 2A); colony structure analysis at the gate level (fig. 2B); F/B ratio (FIG. 2C); structural analysis of the flora at the genus level (fig. 2D); specific abundance analysis of representative bacteria (fig. 2E); shannon (fig. 2F) and simpson analysis (fig. 2G); UniFrac principal coordinates analysis (PCoA) (fig. 2H). N represents Normal, Normal group; m represents Model, Model group; MMF, representing the group given mycophenolate mofetil treatment; HP, representing treatment group given hydrangea paniculata extract.
Fig. 3Lefse analysis results. M represents Model, Model group; (ii) a HP, representing treatment group given hydrangea paniculata extract.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the materials and devices used in the present invention are commercially available unless otherwise specified.
Example 1 c-BSA (cationic Albumin) induces model of Chronic Kidney disease
1. Preparation of antigen-cationized BSA (C-BSA)
Natural bovine serum albumin fraction V: electrophoretically pure, with an isoelectric point of 4.5, Ameresco. Carbodiimide (EDC): beijing chemical company. Anhydrous Ethylenediamine (EDA): analytically pure, Beijing Chemicals, Inc. According to the Border method, 67mL of EDA is added with 500mL of double distilled water, then 350mL of 6M hydrochloric acid is slowly added, the pH value is adjusted to 4.75, the solution is cooled to 25 ℃ on ice, 5g of BSA is dissolved in 25mL of double distilled water, then the solution is slowly added into the EDA solution, stirring is continuously carried out, 1.8g of EDC is added, the reaction is carried out for 2h at the constant temperature of 25 ℃, and then 30mL of acetic acid buffer solution with the pH value of 4.75 is used for stopping the reaction, thus obtaining the C-BSA solution with the improved isoelectric point. Dialyzing with double distilled water at 4 deg.C for 48h (changing water every 8 hr), freeze drying to obtain C-BSA powder with isoelectric Point (PI) of above 8.4, and storing at-70 deg.C.
2 making of model
The experimental animals are healthy female Sprague-Dawley rats with the weight of 160-180 g provided by Beijing Wintoli Hua. After 3 days of acclimatization, randomized into 2 groups: pathological groups were 90, and the nephritis model was replicated with C-BSA; the normal control group was 10, and was normally kept without any treatment.
With reference to the Border method, pre-immunization (prime immunization): 1.5mg of C-BSA was dissolved in 0.5mL of physiological saline, and mixed with 0.5mL of Freund's incomplete adjuvant to prepare "water-in-oil", which was used for subcutaneous multipoint preimmunization, and after 1 week, C-BSA was injected daily into the tail vein. The dose on days 1 to 7 of the tail vein injection was 2.5mg, 3mg, 4mg, 5mg in this order, and thereafter 5mg per day. After 5mg of tail vein, the urine protein of the rat is collected for urine protein determination, and the content of the urine protein of the model group is 5 times higher than that of the normal group, namely the rat which is successfully modeled.
Example 2 preparation of hydrangea paniculata (Hydrangeapaniculata Sieb) extract
Hydrangea paniculata, Saxifragaceae, Stem was purchased from Jinxiu county, the autonomous region of the Guangxi Zhuang nationality in 2014 5 months. The Guangxi Liuzhou forestry bureau identified the samples. The sample specimen has the ID number of 4645, and exists in the institute of medicine of Chinese academy of medicine, Beijing, China at present.
Preparing total coumarin glycosides from hydrangea paniculata extract: pulverizing hydrangea paniculata branch, adding 10 times of water, reflux-extracting for 2 times, each for 3 hr, filtering the extractive solution while it is hot, mixing filtrates, passing the filtrate through macroporous adsorbent resin column (HPD100) with water balance, eluting with 3 times of water with the volume of macromolecular polymer to remove impurities, and discarding the water solution. Desorbing coumarin glycoside with 5 times of 17% ethanol, recovering ethanol from the eluate under reduced pressure, concentrating to obtain extract, and drying to obtain hydrangea paniculata extract. And dissolving the extract with water, passing the solution through a macromolecular polymer column HP2MGL which is well balanced with water in advance, washing with 3 times of the volume of the macromolecular polymer column to remove impurities, desorbing the coumarin glycoside by using 25% ethanol in 3 times of the volume of the macromolecular polymer column, evaporating the eluent under reduced pressure, and drying in vacuum to obtain light brown powder containing various total coumarin glycosides. The content (weight) of hydrangea paniculata coumarin glycoside is determined to be 65% by UV (instrument model JASCO V-650) detection analysis and determination method, which is determined by referring to the ultraviolet spectrophotometry of appendix V B of the 2010 version of Chinese pharmacopoeia.
Preparation and structural identification of monomeric compounds in the total coumarins glycoside of hydrangea paniculata:
taking a hydrangea paniculata total coumarin glycoside sample, carrying out silica gel column (200-300 meshes) chromatographic separation, eluting with chloroform-methanol-water (80: 20: 2) to obtain 8 elution parts (YAA-YAF), and separating out a large amount of white solid, namely a compound YA-1, from YAD. Filtering, and subjecting the filtrate to silica gel column chromatography, Sephadex LH-20 column chromatography, and reversed phase HPLC preparative chromatography to obtain compounds YA-2 and YA-3. YAE precipitates a large amount of white solid, compound YA-5. YAF was further subjected to silica gel column (200-300 mesh) chromatography, eluted with chloroform-methanol-water (80: 20: 2) to give 22 fractions (Frc.1-Frc.11), Frc.9 was further eluted with MPLC (5-50% MeOH gradient), and preparative reverse phase HPLC to give YA-4.
The physicochemical parameters of the compounds are as follows:
1. compound YA-1 (skimmin)
A white solid. ESI-MS M/z 325[ M + H ]]+。1H-NMR(DMSO-d6,500MHz)δ:6.32(1H,d,J＝9.5Hz,H-3),8.00(1H,d,J＝9.5Hz,H-4),7.64(1H,d,J＝8.5Hz,H-5),7.04(1H,s,H-8),7.00(1H,d,J＝8.5Hz，H-6),5.02(1H,d,J＝7.0Hz,H-1′),3.13～3.71(6H,m,H-2′～6′)。13C-NMR(DMSO-d6,125MHz)δ:160.2(C-2),160.2(C-7),155.0(C-9),144.2(C-4),129.4(C-5),113.6(C-6),113.2(C-10),113.1(C-3),103.1(C-8),99.9(C-1′),77.1(C-3′),76.5(C-5′),73.1(C-2′),69.6(C-4′),60.6(C-6′)。
2. Compound YA-2 (6-methoxy coumarin-7-O-beta-D-glucoside (scopolamine))
A white solid. ESI-MS M/z 355[ M + H ]]+。1H-NMR(DMSO-d6,300MHz)δ:7.95(1H,d,J＝9.6Hz,H-4),7.28(1H,s,H-5),7.15(1H,s,H-8),6.32(1H,d,J＝9.6Hz,H-3),5.08(1H,d,J＝7.2Hz,H-1′),3.81(3H,s,OCH3-6),3.15～3.80(6H,m,H-2′～6′)。
3. Compound YA-3 (8-methoxy coumarin-7-O-beta-D-glucoside)
A white solid. ESI-MS M/z 377[ M + Na ]]+。1H-NMR(DMSO-d6,300MHz)δ:7.97(1H,d,J＝9.6Hz,H-4),7.38(1H,d,J＝9.0Hz,H-5),7.17(1H,d,J＝9.0Hz,H-6),6.32(1H,d,J＝9.6Hz,H-3),5.01(1H,d,J＝7.2Hz,H-1′),3.86(3H,s,OCH3-8),3.15～3.80(6H,m,H-2′～6′)。
4. Compound YA-4 (umbelliferone-7-O-beta-D-glucosyl- (1 → 6) -O-beta-D-glucoside)
A white solid. ESI-MS M/z 509[ M + Na ]]+,995[2M+Na]+。1H-NMR(DMSO-d6,400MHz)δ:7.98(1H,d,J＝9.6Hz,H-4),7.64(1H,d,J＝8.8Hz,H-5),7.09(1H,dd,J＝8.8,2.4Hz,H-6),7.06(1H,d,J＝2.4Hz，H-8),6.31(1H,d,J＝9.6Hz,H-3),5.02(1H,d,J＝7.2Hz,H-1′),4.17(1H,d,J＝8.0Hz,H-1″),2.96～4.46(12H,m,H-2′～6′,2″～6″).13C-NMR(DMSO-d6,125MHz)δ:160.2(C-2),160.1(C-7),154.9(C-9),144.2(C-4),129.6(C-5),113.4(C-6),113.3(C-10),113.1(C-3),103.4(C-8),103.5(C-1″),99.9(C-1′),77.0(C-3′),76.7(C-3″),76.4(C-5′),75.6(C-5″),73.5(C-2′),73.1(C-2″),70.1(C-4′),69.5(C-4″),68.6(C-6′),61.1(C-6″)。
5. Compound YA-5 (umbelliferone 7-O-beta-D-apiosyl- (1 → 6) -beta-D-glucoside (hydrangea glucoside))
A white solid. ESI-MS M/z 479[ M + Na ]]+,457[M+H]+。1H-NMR(DMSO-d6,500MHz)δ:7.98(1H,d,J＝9.5Hz,H-4),7.64(1H,d,J＝9.0Hz,H-5),7.01(1H,overlapped,H-6),7.01(1H,brs,H-8),6.32(1H,d,J＝9.5Hz,H-3),5.01(1H,d,J＝7.5Hz,H-1′),4.97(1H,d,J＝6.5Hz,OH-2″),4.79(1H,d,J＝3.0Hz，H-1″),3.89(1H,d,J＝9.5Hz，H-4″),3.86(1H,brd,J＝12.0Hz,H-6′),3.74(1H,dd,J＝6.5,3.0Hz，H-2″),3.58(1H,d,J＝9.5Hz，H-4″),3.59(1H,m,H-5′),3.44(1H,dd,J＝12.0,6.5Hz,H-6′),3.38(1H,overlapped，H-5″),3.29(1H,m,H-3′),25(1H,m,H-2′),3.28(1H,overlapped,H-5″),3.12(1H,m,H-4′)。13C-NMR(DMSO-d6,125MHz)δ:160.2(C-2),113.3(C-3),144.1(C-4),129.5(C-5),113.4(C-6),160.1(C-7),103.3(C-8),154.9(C-9),113.2(C-10),109.3(C-1″),99.9(C-1′),78.7(C-3″),76.4(C-3′),75.9(C-2″),75.5(C-5′),73.3(C-4″),73.0(C-2′),69.8(C-4′),67.5(C-6′),63.2(C-5″)。
At present, 95% of chemical components in HP are identified and clarified, wherein 65% of the chemical components are coumarin derivatives, and the chemical components of the coumarin glycoside derivatives in HP are shown in Table 1 and mainly become skimmin and hydrangea macrophylla.
TABLE 1 chemical composition of coumarin glycoside derivatives in HP
Example 3 animal test for modulating Chronic renal disease intestinal flora
1. 84 SD female mice (Sibefu, Beijing, Biotech, Inc.) are divided into a normal group, a model (M) group, a positive drug mycophenolate mofetil (MMF 20mg/kg) group and an HP (7.5mg/kg, 15mg/kg and 30mg/kg) group, each group contains 14 mice, the administration is performed by intragastric administration for 1 time every day, the administration lasts for 2 months, and all indexes are detected.
2. And (3) observing items:
(1) and (4) checking functions: quantitative determination of urine protein. ② measuring serum creatinine, urine creatinine and serum urea nitrogen.
The results are shown in table 2, the hydrangea paniculata extract can effectively reduce urinary albumin and blood creatinine of the organism, effectively improve renal function, regulate the levels of total cholesterol, high density lipoprotein, low density lipoprotein and triglyceride in serum and regulate lipid metabolism.
(2) And (3) morphological observation: two groups of rats were sacrificed and kidneys were taken, the weight of the rats and the weight of the kidneys were respectively weighed, organ coefficients were calculated, the kidneys were formalin-fixed for later use, and the following observations were performed: fixing renal cortex with 20% formaldehyde, dehydrating with ethanol, embedding in paraffin, making into 3 μm slices, performing HE and PAM-Masson staining, and observing with light microscope.
HE staining results are shown in fig. 1, where severe glomeruli fibrosis occurred in the model group, while the degree of glomerular fibrosis was significantly reduced and lymphocyte infiltration in the renal interstitium was also significantly reduced in the HP-treated rats.
3. Feces Collection, 16S sequencing
The feces from each group of rats were collected, placed in sterile EP tubes, and immediately frozen at-80 ℃.
(1) Extraction and PCR amplification of genomic DNA
Extracting the genome DNA of the fecal sample by using a DNA extraction kit, detecting the purity and concentration of the DNA by using agarose gel electrophoresis, taking a proper amount of sample in a centrifuge tube, and diluting the sample to 1 ng/mu l by using sterile water if necessary. The diluted genome DNA is used as a template, and according to the selection of a sequencing region, a specific primer with Barcode and Takara Ex Taq high-fidelity enzyme of Takara company are used for carrying out PCR, so that the amplification efficiency and the amplification accuracy are ensured. Bacterial diversity identifies the corresponding regions: 16S V3-V4 region (primers 343F and 798R),
343F corresponds to primer sequence 5'-TACGGRAGGCAGCAG-3' (SEQ ID NO: 1) and 798R corresponds to primer sequence 5'-AGGGTATCTAATCCT-3' (SEQ ID NO: 2).
(2) Mixing and purification of PCR products
And (3) detecting a PCR product by using electrophoresis, purifying the PCR product by using magnetic beads after detection, performing two-round PCR amplification by using the purified PCR product as a two-round PCR template, detecting the PCR product by using the electrophoresis again, purifying the PCR product by using the magnetic beads after detection, and performing Qubit quantification on the PCR product after purification. And (5) carrying out equal-quantity sample mixing according to the concentration of the PCR product, and carrying out machine sequencing.
(3) Biological information analysis process
The raw data is in FASTQ format. The original bi-polar sequence was stripped using trimmatic software. The impurity removal parameters are as follows: detecting and truncating the fuzzy base N; and checking the average base quality by a sliding window method, and intercepting the previous high-quality sequence when the quality is lower than 20. And splicing the two-end sequences after the impurity removal by using FLASH software. The splicing parameters are as follows: the minimum Overlap is 10bp, the maximum Overlap is 200bp, and the maximum mismatch rate is 20%. To ensure the accuracy of the result, the method can be used for removing the sequences containing the vague base (ambiguus), the single-base high-repeat region (homologus) and the sequences with too short length. The parameters for accurate impurity removal are as follows: sequences containing N bases were removed, and sequences with a base mass fraction Q20 of at least 75% were retained. Meanwhile, a chimaera sequence in the sequence is detected and removed by using UCHIME. After the sequencing data are preprocessed to generate a high-quality sequence, the sequence is classified into a plurality of OTUs according to the similarity of the sequence by adopting Vsearch software. The parameter that the sequence similarity is greater than or equal to 97% is classified as an OTU unit. Representative sequences of each OTU were picked using the QIIME software package and all representative sequences were annotated against the database. 16S, using Greenengens or silver (version123) database for comparison, using RDP classifier software for species comparison annotation, and keeping the annotation result with a confidence interval larger than 0.7. ITS was aligned using the Unite database. Species alignment annotation blast software was used.
4. Display by sequencing results
A total of 3585 OTUs were identified, and the number of OTUs shared and unique among the different groups was obtained by statistical analysis of the number of tags assigned to OTUs per sample (fig. 2A). With 98 OUT being common to each group. Through colony structure analysis, the inventor finds that the immune nephropathic group is dysbacteriosis, and can find the change of the intestinal flora structure at phylum level, wherein the flora structure mainly comprises firmicutes and bacteroidetes (FIG. 2B). The ratio of firmicutes in the model group was increased and the ratio of bacteroidetes was decreased compared to the normal group, the ratio firmicutes/bacteroidetes (F/B) was significantly increased (P < 0.05). Meanwhile, the HP intervenes, the abundances of firmicutes are reduced, the abundances of bacteroidetes are increased, F/B is reduced compared with a model group, and the effect is better than that of mycophenolate mofetil (figure 2C). The ratio of firmicutes/bacteroidetes (F/B) is often used to reflect the degree of metabolic disease, which well illustrates the great potential of HP in ameliorating metabolic disease.
At the genus level, the inventors showed a representative top30, and found that model group gut flora imbalance, HP reversed gut flora imbalance (fig. 2D). Some representative strains were selected for illustration (FIG. 2E). Lactobacillus and Bifidobacterium are two important intestinal probiotics and play an important role in improving the intestinal microenvironment and maintaining the intestinal health. Compared to the normal group, the abundance of the two genera in the model group was reduced, and the MMF and HP interventions could up-regulate the abundance of the two probiotics, whereas the MMF group differed significantly, especially in bifidobacteria (P < 0.001). Prevotellaceae _ UCG _001 (Prevotella _ UCG _001) is a key genus producing Short Chain Fatty Acids (SCFA), and has trophoblasts and anti-inflammatory effects. The abundance of model group Prevotellaceae _ UCG _001 (Prevotella _ UCG _001) is obviously reduced (p is less than 0.01), and MMF and HP can both up-regulate the abundance of the strain, and HP is superior to MMF. Butyrivibrio (vibrio butyrate) is a butyrate-producing strain and plays an important role in promoting the intestinal barrier function and maintaining the intestinal health. The butyrospirillum abundance in the model group was significantly lower than that in the normal group (p <0.001), and the abundances in the MMF and HP groups were increased, but statistically significant in the mycophenolate group (p < 0.01). Alistipes (cladosporium) are symbionts of the intestinal flora, are limited in number and represent good health of the gastrointestinal tract. Compared with the normal group, the level of the genus is remarkably reduced in the MN group (p <0.01), and both MMF and HP remarkably up-regulate the abundance of the genus (p < 0.01). Escherichia _ Shigella (Shigella) is an enteropathogenic bacterium that usually attacks the colonic epithelium, causing a severe inflammatory response that leads to the death of the colonic epithelium. This inflammation causes dysentery, which is a hallmark of shigella infection. It was found that the MN group Escherichia Shigella was high (p <0.05), while HP intervention significantly reduced the abundance of this strain (p < 0.05). In the model group, Blautia (Blautia) was significantly increased, while in the HP group, the abundance of the genus was significantly decreased. It is worth mentioning that high abundance of Blautia (Blautia) has been shown to be associated with a decrease in early glomerular filtration rate (eGFR), which is consistent with the inventors' results. Consistent with the reported results for CKD, increased abundance of Allobaculum (mycoplasma) and Enterorhabdus (enterobacter) could be observed in the model group (P <0.01), and HP could significantly reduce the level of Allobaculum (mycoplasma) (P < 0.01).
The α -diversity index was used to indicate ecological diversity in microbial communities, including shannon and simpson analysis (fig. 2F, 2G). The inventor finds that the microbial diversity of the chronic kidney disease model group is changed remarkably, and the microbial diversity gradually trends to be normal through HP intervention. The UniFrac principal axis analysis (PCoA) showed significant microflora in all treatment groups (fig. 2H). The inventors could see that the flora composition of the HP group is concentrated near the normal group, indicating that the flora structure difference between the two is small, which reflects that HP can regulate the normalization of intestinal flora to a certain extent.
By Lefse analysis (fig. 3), the key difference between the HP administration group and the model group was found to be characteristic of the genus.
The inventors found that the genus differentially belonging to Prevotella _ UCG _001 (Prevoteriaceae _ UCG001 genus), Rikenella (Hodgiaceae), Alisipes (Allophyllaceae), Proteobacteria (Proteobacteria), Turcibacter (Zuricobacter), Clostridiales _ vadinBB60_ group (Clostridiales vaBB 60 genus), Ruminococcaceae _ UCG _010 (Ruminococcaceae _ UCG _010 genus), Clostridiaceae _1 (Clostridium _1) had high abundance in HP administration group, while Prevotella _1 (Proteobacteria 1), Prevotella _9 (Proteobacteria _9), Lachnospiraceae (Lachnospiraceae), Streptococcus (Streptococcus), Allobacillus (Proteobacteria), Fuocibacter (Fusobacterium 008), Ruminococcaceae _ UCG (Corynebacterium), and Ruminococcal (Corynebacterium) had high abundance in Corynebacterium model. The genus pleiotrophicus can be used as a biomarker for disease diagnosis and treatment.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The application of the hydrangea paniculata extract in preparing the medicine for preventing or treating chronic nephropathy is characterized in that the hydrangea paniculata extract is extracted by reflux extraction, macroporous resin ion exchange and reduced pressure distillation, and the hydrangea paniculata extract is used for preventing or treating chronic nephropathy by regulating intestinal flora.
2. The use of claim 1, wherein 65% by weight of the hydrangea paniculata extract is coumarin glycosides.
3. The use of claim 2, wherein the coumarin glycoside compounds comprise skimmin, hydrangea glycoside, scopolamine, 8-methoxycoumarin-7-O- β -D-glucoside and umbelliferone-7-O- β -D-glucosyl- (1 → 6) -O- β -D-glucoside as main ingredients.
4. The application of claim 3, wherein the specific method for extracting the coumarins glycosides from the hydrangea paniculata extract comprises:
(1) pulverizing hydrangea paniculata branches, adding 10 times of water, reflux-extracting for 2 times, each time for 3 hr, filtering the extractive solution while it is hot, and mixing filtrates;
(2) passing the filtrate through macroporous adsorbent resin column (HPD100), eluting with 3 times of water of macromolecular polymer volume to remove impurities, and discarding water solution; desorbing coumarin glycoside with 5 times of 17% ethanol, recovering ethanol from eluate under reduced pressure, concentrating to obtain extract, and drying to obtain hydrangea paniculata extract;
(3) dissolving the extract with water, passing the solution through a macromolecular polymer column HP2MGL, eluting with 3 times of water to remove impurities; desorbing coumarin glycoside with 3 times of macromolecular polymer volume of 25% ethanol, evaporating eluate under reduced pressure, and vacuum drying to obtain light brown powder containing various total coumarin glycosides.
5. The use of claim 4, wherein the hydrangea paniculata extract improves the intestinal flora structure, resulting in a decrease in firmicutes abundance, an increase in bacteroidetes abundance, and a decrease in the firmicutes/bacteroidetes ratio.
6. The use of claim 5, wherein the hydrangea paniculata extract reverses a disturbance in the gut flora comprising Bifidobacterium, Vibrio butyricum, Shigella, Enterobacter, Lactobacillus, Proteus _9, Proteus _ NK4A136, Proteus 1, Bacteroides, Prevotella _ UCG001, Ruminococcaceae _ UCG005, Zuricobacter, Mycoplasma, Ruminococcus 1, Scutellaria, Blauteria, Ruminococcaceae UCG014, Desulfornia, helicobacter, Rosemonium, Streptococcus, Prevoteriaceae _ 3B31, NK Proteus UCG001, Clostridium, Rooiticillus, ParaSainta, Clostridium, Klitenbergeridae, Klitella R7, Oscillatoria, Anaerobiospirillum, Spirochaetaceae G005, and Proteus, Clostridium and Youyoba.
7. The application of the hydrangea paniculata extract in the medicines for regulating intestinal flora is characterized in that the hydrangea paniculata extract is extracted according to the methods of reflux extraction, macroporous resin ion exchange and reduced pressure distillation.
8. The use of claim 7, wherein the hydrangea paniculata extract improves gut flora architecture, reverses gut flora imbalance, and modulates gut microbial diversity.
9. The application of the biomarker of chronic kidney disease in the preparation of a kit or a reagent for detecting chronic kidney disease is characterized in that the biomarker of chronic kidney disease at least comprises any one or more groups of the following microbial flora: prevoteriaceae _ UCG001, Hodgsoniaceae, Arthrobacter, Proteobacteria, Zuricobacter, order VadinBB60, Ruminococcaceae _ UCG _010, Clostridium _1, Proteobacteria _9, Lachnospiraceae, Streptococcus, Mycoplasma, Mycobacterium, Ruminococcaceae _ UCG _008, anaerobic Corynebacterium, Bifidobacterium, Vibrio butyricum, Shigella, Enterobacter, lactobacillus, Bacteroides, Prevotella, Ruminococcaceae _ UCG005, Ruminococcus 1, Blattella, Ruminococcaceae UCG014, Desulvronas, helicobacter, Roseburia, Prevotetaceae _ NK3B31, Clostridium, Rooibos, ParaSaturella, Clostridium, Klitenberg R7, Oscillatoria, anaerobic stick, Clostridium, Youngia.
10. Use of a biomarker of chronic kidney disease comprising at least a set of any one or more of the following microbial flora: prevoteriaceae _ UCG001, Hodgsoniaceae, Arthrobacter, Proteobacteria, Zuricobacter, order VadinBB60, Ruminococcaceae _ UCG _010, Clostridium _1, Proteobacteria _9, Lachnospiraceae, Streptococcus, Mycobacteria, Ruminococcaceae _ UCG _008, anaerobic Corynebacterium, Bifidobacterium, Vibrio butyricum, Shigella, Enterobacter, lactobacillus, Bacteroides, Prevotella, Ruminococcaceae _ UCG005, Ruminococcus 1, Blattella, Ruminococcaceae UCG014, Desulvronas, helicobacter, Roseburia, Prevotetaceae _ NK3B31, Clostridium, Rooibos, ParaSaturella, Clostridium, Klitenberg R7, Oscillatoria, anaerobic stick, Clostridium, Youngia.
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