WO2023098764A1 - 乳双歧杆菌bl-99在预防和/或改善胃炎中的应用 - Google Patents
乳双歧杆菌bl-99在预防和/或改善胃炎中的应用 Download PDFInfo
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- WO2023098764A1 WO2023098764A1 PCT/CN2022/135672 CN2022135672W WO2023098764A1 WO 2023098764 A1 WO2023098764 A1 WO 2023098764A1 CN 2022135672 W CN2022135672 W CN 2022135672W WO 2023098764 A1 WO2023098764 A1 WO 2023098764A1
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- bifidobacterium lactis
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- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/531—Lactis
Definitions
- the present invention relates to the new application of Bifidobacterium lactis (Bifidobacterium lactis) BL-99 (preservation number CGMCC No.15650), specifically, the present invention relates to the preparation of Bifidobacterium lactis BL-99 for preventing and/or Use in a composition for improving gastritis.
- Chronic gastritis is a common disease of the digestive system, often combined with peptic ulcer, and is one of the precancerous diseases of gastric cancer.
- the clinical symptoms of patients are epigastric discomfort, fullness, loss of appetite, acid reflux, abdominal distension and pain, etc. Some patients may also have systemic mental symptoms such as fatigue, weight loss, anxiety, and depression. Relevant clinical studies have proved that reversing the progression of chronic gastritis is also the key to preventing gastric cancer.
- serum pepsinogen pepsinogen, PG
- gastrin-17 gastrin-17
- Serum PGI, PGII, PGI/PGII ratio combined with anti-H.pylori antibody detection is helpful for stratified risk management of chronic gastritis.
- the PG level reflects the functional status of the gastric mucosa.
- the levels of PGI and PGII decrease, and the PGI level decreases more significantly, so the ratio of PGI/PGII decreases accordingly.
- PG determination helps to judge the extent of atrophy.
- the ratio of PGI and PGI/PGII decreased in patients with gastric body atrophy, and the level of serum gastrin-17 increased; in patients with atrophy of gastric antrum, the level of serum gastrin-17 decreased, and the ratio of PGI and PGI/PGII was normal; in patients with total gastric atrophy, both Both decreased.
- the PGIlevel ⁇ 70g/L and the PGI/PGIIratio ⁇ 3.0 are used as the diagnostic cut-off value of atrophic gastritis, and this is used as the screening standard for gastric cancer high-risk groups. It is widely used in the screening of gastric cancer risk in Europe and Japan; the standard of PGI level ⁇ 70g/L and PGI/PGII ⁇ 7.0 is often used in the screening of high-incidence areas of gastric cancer in China, and there is still a lack of follow-up data of large samples to support it.
- Serum PG combined with serum anti-H.pylori antibody detection can divide the population into four groups: A, B, C, and D.
- the incidence of gastric cancer in different groups is different, which is a valuable predictor of gastric cancer risk.
- Japan uses this As a stratification method (ABCD method) of gastric cancer risk, a corresponding inspection strategy was formulated.
- ABCD method stratification method
- the PGI/PGII ratio is negatively correlated with OLGA staging, the lower the ratio, the higher the stage, and the PGI/PGII ratio ⁇ 3.0 can be used to distinguish low-risk and high-risk OLGA stages, with a sensitivity of 77% and a specificity of 85%.
- the positive predictive value was 45%, and the negative predictive value was as high as 96%.
- H. pylori infection can increase the levels of PGI and PGII, especially PGII, so the ratio of PGI/PGII decreases.
- One object of the present invention is to provide a new application of Bifidobacterium lactis.
- the present invention provides a new application of Bifidobacterium lactis (Lactobacillus paracasei) BL-99 strain, which has been preserved in China Microorganisms on December 18, 2017 The General Microorganism Center of the Preservation Management Committee, the deposit number is CGMCC No.15650.
- Bifidobacterium lactis BL-99 bacterial strain is the biological material that has been announced in the patent document CN 110964653 A. Research in the prior art shows that Bifidobacterium lactis BL-99 is a safe edible probiotic, which is resistant to gastric acid and intestinal juice.
- Bifidobacterium lactis BL-99 has the functions of enhancing intestinal immunity, improving the integrity of intestinal barrier, and inhibiting the production of intestinal inflammatory factors.
- Bifidobacterium lactis (Bifidobacterium lactis) (i.e. Bifidobacterium lactis BL-99) with the preservation number CGMCC No. 15650 has the effect of preventing and/or improving gastritis, specifically One or more aspects of the following effects can be manifested: improving the symptoms of functional dyspepsia; increasing the index levels of PGI and PGI/PGII; regulating gastric acid secretion; and/or reducing the expression of inflammatory factors IL-6 and TNF- ⁇ .
- the Bifidobacterium lactis BL-99 of the present invention has good potential for preventing and/or improving stomachs such as chronic gastritis and atrophic gastritis (gastric mucosal atrophy associated with gastritis).
- the present invention provides an application of Bifidobacterium lactis in the preparation of a composition for preventing and/or improving gastritis, wherein the Bifidobacterium lactis is the lactobacillus with preservation number CGMCC No.15650.
- Bifidobacterium ie Bifidobacterium lactis BL-99.
- the prevention and/or improvement of gastritis includes one or more aspects of the following effects:
- said raising the index level of PGI and/or PGI/PGII comprises increasing pepsinogen I (PG1) secretion and/or improving PG1 /PGII ratio.
- the prevention and/or improvement of gastritis includes:
- the “improvement” mentioned in the present invention includes the relief of related symptoms.
- the composition in the application of the Bifidobacterium lactis BL-99 of the present invention, includes a food composition, a feed composition or a pharmaceutical composition.
- the Bifidobacterium lactis in the application of the Bifidobacterium lactis BL-99 of the present invention, can be used to prepare the composition in the form of a solid or liquid bacterial preparation.
- the Bifidobacterium lactis in the application of the Bifidobacterium lactis BL-99 of the present invention, can be used to prepare the composition in the form of live bacteria or inactivated bacteria.
- the composition in the application of Bifidobacterium lactis BL-99 of the present invention, is a pharmaceutical composition.
- the pharmaceutical composition also includes pharmaceutically acceptable auxiliary materials, which include excipients, diluents, fillers and/or absorption promoters.
- the application amount of the Bifidobacterium lactis is 1.0 ⁇ 10 3 CFU/day-1.0 ⁇ 10 12 CFU/day, preferably 1.0 ⁇ 10 7 CFU/day-1.0 ⁇ 10 11 CFU/day.
- the composition is a food composition.
- the food may be fermented milk products, cheese, milk-containing beverage, solid beverage, compressed candy, milk flakes, ice cream or milk powder.
- the composition in the present invention may also include common material components in the field.
- an appropriate amount of excipients may be included, and the adjuvants may be excipients, diluents, fillers and/or absorption promoters and the like.
- Bifidobacterium lactis of the present invention can be produced according to the food containing Bifidobacterium lactis in the prior art.
- the composition can take different forms according to the needs of the recipient. For example powders, lozenges, granules, microcapsules and/or liquid preparations and the like.
- the present invention also provides a composition for preventing and/or improving gastritis, which includes an effective amount of Bifidobacterium lactis with deposit number CGMCC No.15650.
- the composition for preventing and/or improving gastritis of the present invention includes food composition, feed composition or pharmaceutical composition.
- the present invention also provides a method for preventing and/or improving gastritis, the method comprising administering an effective amount of Bifidobacterium lactis with deposit number CGMCC No. 15650 to the subject.
- the prevention and/or improvement of gastritis includes one or more aspects of the following effects:
- the amount of the Bifidobacterium lactis administered to the subject is 1.0 ⁇ 10 3 CFU/day-1.0 ⁇ 10 12 CFU/day, preferably 1.0 ⁇ 10 7 CFU/day-1.0 ⁇ 10 11 CFU/day.
- the present invention provides a new application of bifidobacterium lactis BL-99 in the preparation of a composition for preventing and/or improving gastritis.
- the present invention proves through experiments that Bifidobacterium lactis BL-99 can improve the symptoms of functional dyspepsia; increase the index levels of PGI and PGI/PGII; regulate gastric acid secretion; and/or reduce the expression of inflammatory factors IL-6 and TNF- ⁇ . Therefore, it can be shown that the Bifidobacterium lactis BL-99 of the present invention has good potential for preventing and/or improving stomachs such as chronic gastritis and atrophic gastritis (gastric mucosal atrophy associated with gastritis).
- FIG. 1 shows that Bifidobacterium lactis BL-99 significantly increased pepsinogen I (PG1) secretion (panel A) and increased the PG1/PGII ratio (panel B).
- PG1 pepsinogen I
- Figure 2 shows that Bifidobacterium lactis BL-99 significantly reduces the secretion of G-17 in people with hyperacidity.
- Figure 3 shows that Bifidobacterium lactis BL-99 significantly increases the secretion of G-17 in people with hypochlorhydria.
- Figure 4 shows that Bifidobacterium lactis BL-99 significantly reduces the expression of inflammatory factor IL-6.
- Figure 5 shows that Bifidobacterium lactis BL-99 significantly reduces the expression of inflammatory factor TNF- ⁇ .
- each original reagent material can be obtained commercially, and the experimental methods without specific conditions are conventional methods and conventional conditions well known in the art, or according to the conditions suggested by the instrument manufacturer.
- Probiotic BL-99 solid drink (BL-99 powder 3.34%, maltodextrin 96.66%) low dose 1 ⁇ 10 10 CFU/day (group E), high dose 5 ⁇ 10 10 CFU/day (group F) BL -99 bacteria powder 16.67%, maltodextrin 83.33%.
- group D Probiotic BL-99 solid drink
- Inclusion criteria One or more symptoms, namely postprandial fullness, early satiety, epigastric pain, epigastric burning sensation, postprandial nausea or vomiting, belching, epigastric flatulence, acid reflux, burning sensation, Retrosternal pain or discomfort, etc., the course of disease is more than 6 months, and symptoms have occurred in the past three months.
- Exclusion criteria patients with gastric Helicobacter pylori infection, patients with acute diarrhea, patients with indigestion caused by severe organic diseases, patients with severe systemic diseases such as cardiovascular, liver, kidney and hematopoietic system, and those who are too weak to accept the test.
- Crowd 300 people, divided into 3 groups, placebo group (group D), low dose group (group E, 10 billion CFU), high dose group (group F, 50 billion CFU).
- the experimental period of each batch of people was 11 weeks, including a 7-day emptying period, an 8-week bacterial powder drinking period, and a 2-week follow-up period.
- the subjects take it with warm water at 45°C within 1 hour after lunch or dinner, and drink 1 bag per day.
- ELISA kit was used to detect the concentration of gastrin (G-17) in serum, and immunoturbidimetric kit was used to detect the concentration of pepsinogen I (PGI) and pepsinogen II (PGII) in serum.
- the ratio of PGI to PGII (PGR) was calculated, and the gastric digestive ability index was detected to judge the efficacy of BL-99 on the gastric digestive ability of the population.
- Serum cytokines IL-6 and TNF- ⁇ were detected by ELISA kit (excell-bio).
- Gender data are expressed as % of the population; body mass index (BMI) is weight (kg) divided by height (m) squared.
- Pepsinogens are derived from gastric mucosal cells and can be divided into two groups according to immunohistochemistry: pepsinogen I (PG1) and pepsinogen II (PGII), mainly secreted into the gastric cavity, only 1% Left and right PGs circulate in the blood.
- PGI is mainly secreted by the bottom mucosal cells
- PGII is mainly secreted by chief cells, duodenal cells and pyloric glands.
- Bifidobacterium lactis BL-99 helps to increase the index levels of PGI and PGI/PGII, and has good potential for preventing and improving gastric diseases such as chronic gastritis and atrophic gastritis.
- Gastrin is one of the gastrointestinal peptide hormones, which can promote the release of histamine from pheochromocytoma and stimulate the secretion of gastric acid. If the G-17 value is higher than 15, it will aggravate the occurrence of gastritis, and it is also the precursor of gastric cancer transformation. One of the indicators. Conversely, if G-17 ⁇ 1, gastric acid secretion is too low, manifested as insufficient gastric motility, weakened digestion ability, early satiety, upper abdominal fullness and other symptoms. After 2 months of intervention with Bifidobacterium lactis BL-99 (Group E, Group F), the G-17 value was detected during the period. Please refer to Figure 2 and Figure 3 for the results.
- Gastric acid secretion disorder is one of the main causes of chronic gastritis, and the use of proton pump inhibitors such as omeprazole to inhibit gastric acid secretion is also the most common clinical treatment for chronic gastritis.
- proton pump inhibitors such as omeprazole
- omeprazole to inhibit gastric acid secretion
- more and more studies have shown that long-term use of proton pump inhibitors can lead to abnormal absorption of nutrients (vitamins, minerals, etc.) in the gastrointestinal tract, damage to the gastrointestinal mucosa, increased risk of kidney and cardiovascular diseases, and bone metabolism. exception etc.
- the use of proton pump inhibitors can easily cause the disturbance of intestinal flora, and it is also related to the occurrence of Alzheimer's disease in the elderly. Based on this, the development of ingredients that can help reduce the dosage of proton pump inhibitors, alleviate side effects, and even replace proton pump inhibitors has extensive clinical significance.
- the present invention finds that the probiotic strain Bifidobacterium lactis BL-99 has the effect of bidirectionally regulating gastric acid secretion.
- Gastric precancer is a pathological concept, including intestinal metaplasia and dysplasia, which is an important stage in the transformation process from normal gastric tissue like gastric cancer.
- Gastric cancer has a high mortality rate because it was discovered after the transformation, so "inflammation and cancer transformation" is also a hot spot of clinical attention.
- inflammation and cancer transformation is also a hot spot of clinical attention.
- pro-inflammatory factors in this process has also been continuously defined.
- Interleukin-6 has dual functions of pro-inflammation and anti-inflammation in the body. Under the stimulation of inflammatory response, IL-6 can be expressed in large quantities, prompting the release of monocytes and neutrophils, and in the A large number of inflammatory sites gather; TNF- ⁇ is also an important mediator in the inflammatory process, mainly produced in activated monocytes and macrophages, and its large expression will also promote the release of other inflammatory factors. With the intensification of gastric mucosal inflammatory response, a large number of inflammatory factors are released and infiltrate gastric cells, accelerate the degree of mucosal atrophy, invade gastric mucosa, reduce the ability of intracellular glands to secrete PG, and affect gastrointestinal dynamics. Inflammatory factors IL-6, TNF- ⁇ interact with PGI, PGII, G-17, etc., and jointly affect gastric function.
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Abstract
乳双歧杆菌BL-99在预防和/或改善胃炎中的应用。本发明提供了一种乳双歧杆菌(Bifidobacterium lactis)在制备用于预防和/或改善胃炎的组合物中的应用,其中,所述乳双歧杆菌为保藏编号CGMCC No.15650的乳双歧杆菌。所述乳双歧杆菌可预防和/或改善慢性胃炎,和/或预防和/或改善胃炎相关的胃粘膜萎缩。
Description
本发明是关于乳双歧杆菌(Bifidobacterium lactis)BL-99(保藏编号CGMCC No.15650)的新应用,具体而言,本发明是关于乳双歧杆菌BL-99在制备用于预防和/或改善胃炎的组合物中的应用。
慢性胃炎是消化道系统常见病,常合并消化性溃疡,为胃癌癌前疾病之一。近年来,生活水平的提高及工作压力增加,使慢性胃炎患病率逐年上升。患者的临床症状为上腹不适、饱胀、食欲减退、反酸、腹胀痛等,部分患者还可有乏力、消瘦、焦虑、抑郁等全身精神症状。相关临床研究证明,逆转慢性胃炎进展也是预防胃癌发生的关键。
血清胃蛋白酶原(pepsinogen,PG)Ⅰ、Ⅱ以及促胃液素-17(gastrin-17)的检测可能有助于判断有无胃黏膜萎缩和程度。血清PGⅠ、PGⅡ、PGⅠ/PGⅡ比值联合抗H.pylori抗体检测有助于对于慢性胃炎风险分层管理。
PG水平反映胃黏膜的功能状态,当胃黏膜出现萎缩时,PGⅠ和PGⅡ水平下降,PGⅠ水平下降更明显,因而PGⅠ/PGⅡ比值随之降低。PG测定有助于判断萎缩的范围。胃体萎缩者PGⅠ、PGⅠ/PGⅡ比值降低,血清促胃液素-17水平升高;胃窦萎缩者,血清促胃液素-17水平降低,PGⅠ、PGⅠ/PGⅡ比值正常;全胃萎缩者则两者均降低。通常以PGⅠ水平≤70g/L且PGⅠ/PGⅡ比值≤3.0作为萎缩性胃炎的诊断临界值,并以此作为胃癌高危人群筛查的标准。在欧洲和日本广泛用于胃癌风险的筛查;国内胃癌高发区筛查常采用PGⅠ水平≤70g/L且PGⅠ/PGⅡ≤7.0的标准,目前尚缺乏大样本的随访数据加以佐证。
多项研究证明,血清PG检测有助于胃癌高危人群的风险分层,PG检测诊断萎缩者,以及PG检测虽诊断萎缩阴性但PGⅠ/PGⅡ比值较低者,有较高的胃癌风险,应进一步进行胃镜检查。血清PG联合血清抗H.pylori抗体检测可将人群分为A、B、C、D 4组,不同组别其胃癌的发生率不同,是一项有价值的胃癌风险的预测指标,日本以此作为胃癌风险的分层方法(ABCD法),制定相应的检查策略。最 近国内的研究结果显示,PG、促胃液素-17和抗H.pylori抗体联合检测可对胃癌发生风险加以分层,辨识出高危个体进行胃镜检查。
PGⅠ/PGⅡ比值与OLGA分期呈负相关,比值越低,分期越高,以PGⅠ/PGⅡ比值≤3.0为界可区别低危和高危OLGA分期,其敏感度为77%,特异度为85%,阳性预测值为45%,阴性预测值高达96%。H.pylori感染可致PGⅠ、PGⅡ水平升高,尤其是PGⅡ更明显,因此PGⅠ/PGⅡ比值下降,根除H.pylori后则PGⅠ、PGⅡ水平下降,PGⅠ/PGⅡ比值上升。
发明内容
本发明的一个目的在于提供一种乳双歧杆菌的新用途。
根据本发明的具体实施方案,本发明提供了乳双歧杆菌(Lactobacillus paracasei)BL-99菌株的新用途,乳双歧杆菌BL-99菌株已于2017年12月18日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.15650。乳双歧杆菌BL-99菌株是已在专利文献CN 110964653 A中公布的生物材料。现有技术研究表明,乳双歧杆菌BL-99是一株食用安全的益生菌,其具有耐胃酸和耐肠液性能。此外,乳双歧杆菌BL-99具有提升肠道免疫力、提高肠道屏障完整性、抑制肠道炎症因子产生等作用。
本案发明人在研究中通过实验发现,保藏编号CGMCC No.15650的乳双歧杆菌乳双歧杆菌(Bifidobacterium lactis)(即乳双歧杆菌BL-99)具有预防和/或改善胃炎的功效,具体可表现以下功效的一种或多种方面:改善功能性消化不良症状;提升PGI和PGI/PGII的指标水平;调节胃酸分泌量;和/或降低炎症因子IL-6、TNF-α的表达。从而可表明,本发明的乳双歧杆菌BL-99对于预防和/或改善慢性胃炎、萎缩性胃炎(胃炎相关的胃粘膜萎缩)等胃部具有良好的潜力。
从而,本发明提供了一种乳双歧杆菌(Bifidobacterium lactis)在制备用于预防和/或改善胃炎的组合物中的应用,其中,所述乳双歧杆菌为保藏编号CGMCC No.15650的乳双歧杆菌(即乳双歧杆菌BL-99)。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述预防和/或改善胃炎包括以下功效的一种或多种方面:
改善功能性消化不良症状;
提升胃蛋白酶原I(PG1)和/或PGI/PGII的指标水平;
调节胃酸分泌量;和/或
降低炎症因子IL-6和/或TNF-α的表达。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述提升PGI和/或PGI/PGII的指标水平包括增加胃蛋白酶原I(PG1)分泌和/或提高PG1/PGII比值。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述预防和/或改善胃炎包括:
预防和/或改善慢性胃炎,和/或
预防和/或改善胃炎相关的胃粘膜萎缩。
本发明中所述的“改善”包括对相关症状的缓解。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述组合物包括食品组合物、饲料组合物或药品组合物。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述乳双歧杆菌可以固态或液态菌制剂的形式用于制备所述组合物。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述乳双歧杆菌可以活菌或灭活菌的形式用于制备所述组合物。
根据本发明的一些具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述组合物为药品组合物。所述药品组合物还包括药学上可接受的辅料,所述辅料包括赋型剂、稀释剂、填充剂和/或吸收促进剂。
根据本发明的具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述乳双歧杆菌的应用量为1.0×10
3CFU/天-1.0×10
12CFU/天,优选为1.0×10
7CFU/天-1.0×10
11CFU/天。
根据本发明的一些具体实施方案,本发明的乳双歧杆菌BL-99的应用中,所述组合物为食品组合物。其中,所述食品可以为发酵乳制品、乳酪、含乳饮料、固体饮料、压片糖果、奶片、冰淇淋或乳粉。
根据本发明的具体实施方案,本发明中所述组合物还可包括所属领域中的常规用料组分。例如,对于药品组合物,可包括适量的辅料,所述辅料可以为赋型剂、稀释剂、填充剂和/或吸收促进剂等。对于食品组合物,本发明的乳双歧杆菌可以按 照现有技术中含乳双歧杆菌的食品进行生产。所述组合物可根据受施予者的需要,而采用不同形态。例如粉剂、锭剂、造粒、微胶囊和/或液体制剂等。
另一方面,本发明还提供了一种用于预防和/或改善胃炎的组合物,其包括有效量的保藏编号CGMCC No.15650的乳双歧杆菌。
根据本发明的具体实施方案,本发明的用于预防和/或改善胃炎的组合物包括食品组合物、饲料组合物或药品组合物。
另一方面,本发明还提供了一种预防和/或改善胃炎的方法,该方法包括给予受试者有效量的保藏编号CGMCC No.15650的乳双歧杆菌。
根据本发明的具体实施方案,所述预防和/或改善胃炎包括以下功效的一种或多种方面:
改善功能性消化不良症状;
提升胃蛋白酶原I(PG1)和/或PGI/PGII的指标水平;
调节胃酸分泌量;和/或
降低炎症因子IL-6和/或TNF-α的表达。
根据本发明的具体实施方案,所述乳双歧杆菌给予受试者的量为1.0×10
3CFU/天-1.0×10
12CFU/天,优选为1.0×10
7CFU/天-1.0×10
11CFU/天。
本发明提供了乳双歧杆菌BL-99在制备用于预防和/或改善胃炎的组合物中的新应用。本发明通过实验证明乳双歧杆菌BL-99可改善功能性消化不良症状;提升PGI和PGI/PGII的指标水平;调节胃酸分泌量;和/或降低炎症因子IL-6、TNF-α的表达。从而可表明,本发明的乳双歧杆菌BL-99对于预防和/或改善慢性胃炎、萎缩性胃炎(胃炎相关的胃粘膜萎缩)等胃部具有良好的潜力。
图1显示乳双歧杆菌BL-99显著增加胃蛋白酶原I(PG1)分泌(图片A)、提高PG1/PGII比值(图片B)。
图2显示乳双歧杆菌BL-99显著降低胃酸过高人群G-17的分泌。
图3显示乳双歧杆菌BL-99显著提升胃酸不足人群G-17的分泌。
图4显示乳双歧杆菌BL-99显著降低炎症因子IL-6的表达。
图5显示乳双歧杆菌BL-99显著降低炎症因子TNF-α的表达。
为了对本发明的技术特征、目的和有益效果有更加清楚的理解,现结合具体实施例及对本发明的技术方案进行以下详细说明,应理解这些实例仅用于说明本发明而不用于限制本发明的范围。
实施例中,各原始试剂材料均可商购获得,未注明具体条件的实验方法为所属领域熟知的常规方法和常规条件,或按照仪器制造商所建议的条件。
实施例1
1.实验材料与方法
1.1菌种和受试样品
益生菌BL-99固体饮料(BL-99菌粉3.34%,麦芽糊精96.66%)低剂量1×10
10CFU/天(E组),高剂量5×10
10CFU/天(F组)BL-99菌粉16.67%,麦芽糊精83.33%。参照固体饮料配方,将其中的菌粉用同等剂量的麦芽糊精替代后(即全部为麦芽糊精)作为安慰剂组(D组)。
1.2受试对象
纳入标准:具有一项或多项症状,即餐后饱胀感、早饱、上腹疼痛、上腹灼烧感、餐后恶心或呕吐、嗳气、上腹胀气、反酸、灼烧感、胸骨后疼痛或不适等,病程超过6个月,近三个月有症状发作。
排除标准:胃幽门螺旋杆菌感染患者,急性腹泻者,严重器质性病变引起的消化不良者,合并有心血管、肝、肾和造血系统等严重全身性疾病患者,体质虚弱无法接受试验者。
1.3仪器与设备
Sigma 3K30低温高速离心机,德国Satorious公司;MS2漩涡振荡仪,德国IKA公司;万分之一电子天平(TP-214),美国丹佛仪器公司,气相色谱仪(Agilent GC-8860),气相色谱质谱联用仪(GC/MS,Agilent 7200-7890B Accurate-Mass Q-TOF)。
1.4.实验研究方案设计
人群:300人,分成3组,安慰剂组(D组)、低剂量组(E组,100亿CFU)、高剂量组(F组,500亿CFU)。每批人群实验周期11周,包括7天排空期、8周菌粉饮 用期和2周跟踪期。菌粉置于2-8℃冰箱存放,受试者在饮用期内每日午饭或晚饭后1h以内45℃温水冲服,每天饮用1袋。分别在排空期结束(V1期)、饮用期第四周结束(V2期)和第八周结束(V3期)、跟踪期两周(V4期)结束时收集受试者新鲜便样和血液样本,并由医生进行临床症状评分(如表1所示),测试开始至结束,受试者每天记录饮食习惯、消化系统改善评分、肠道健康状况评分、排便习惯满意度评分。辅助药物干预组实验期间不改变原来的药物服用习惯及个人生活习惯。在整个实验过程中,受试者不可饮用任何发酵食品(包括泡菜、酸菜、腐乳、豆豉、奶酪、活性发酵乳饮料等)和其他益生菌产品。
表1临床症状评分
1.5检测指标
1.5.1血清样本分析
受试者全血5mL,在室温放置2h左右,3500r/min,离心15分钟,血清于-80℃保存,待测。
采样当天空腹采血,贴好标签,在规定时间内送到医学检测中心,运输过程避免激烈震荡。
1.5.2胃部消化能力分析
采用ELISA试剂盒检测血清中胃泌素(G-17)的浓度,采用免疫比浊法试剂盒检测血清中胃蛋白酶原Ⅰ(PGⅠ)和胃蛋白酶原Ⅱ(PGⅡ)的浓度,最后通过计算法计算PGⅠ与PGⅡ的比值(PGR),通过对胃部消化能力指标检测,以判断BL-99对人群胃 部消化能力的功效研究。
1.5.3细胞因子分析
采用ELISA试剂盒(excell-bio)检测血清中细胞因子IL-6和TNF-α。
2.数据统计分析
利用SPSS 20.0、GraphPad Prism 8进行统计分析。为了描述受试者的社会人口学特征,使用描述性统计(百分比、平均值和标准差)。组内比较分别采用独立t检验、重复测量方差分析(ANOVA)检验或曼-惠特尼(Mann-Whitney)检验方法,多组间比较采用克鲁斯卡尔-沃利斯(Kruskal-Wallis)检验,计数资料采用卡方检验和卡方趋势检验。P<0.05具有显著性差异和统计学意义。
3.实验结果
3.1人群实验完成情况统计
实验共招募消化不良295人,脱组37人,完成258人,脱组率14.3%。
3.2人群基本信息统计
对受试者的基本信息进行统计,人群各组间进行比较无显著性差异(如表2所示)。
表2功能性消化不良受试者基本信息统计
注:数据以平均值±标准差(Mean±SD)或中位数(四分位数范围)表示。性别数据以人数所占比例%表示;身体质量指数(BMI)是体重(公斤)除以身高(米)的平方。
3.3BL-99对消化不良总人群临床症状积分的影响
对受试者饮用菌粉前后的临床症状积分进行统计分析,结果如表3所示,与V1期相比,饮用BL-99 8周(V3期)后,三组受试者腹痛、腹胀、早饱、恶心呕吐等临 床症状积分均降低。干预八周后(V3期),与安慰剂组相比,低剂量组和高剂量组腹胀、早饱、反酸、腹部灼烧、嗳气和恶心呕吐临床症状积分均降低,BL-99高剂量组能显著改善消化不良腹部灼烧、嗳气症状(P<0.05),且临床症状总分显著性下降(P<0.05)。与低剂量组相比,高剂量组显著改善嗳气症状。表明饮用BL-99菌粉可以显著改善功能性消化不良症状,且饮用8周高剂量的BL-99效果最佳。
表3功能性消化不良受试者临床症状评分变化情况
胃蛋白酶原(Pepsinogens,PG)来源于胃粘膜细胞,依据免疫组化可以分为两个组:胃蛋白酶原I(PG1)和胃蛋白酶原II(PGII),主要分泌到胃腔,只有1%左右的PG在血液中循环。PGI主要是由底部黏膜细胞分泌,PGII主要由主细胞、十二指肠细胞和幽门腺分泌。在慢性胃炎和萎缩性胃炎的前期筛查过程,PGI指标升高,PG1/PGII下降是临床报道最多的生物指标,包含对胃酸状态和胃幽门螺旋杆菌清除的诊断上。
对PGI<50的功能性消化不良受试者进行干预研究,结果请参见图1。实验结果表明,乳双歧杆菌BL-99(E组、F组)连续干预2个月后,PGI指标显著增加,达到正常水平,并且呈剂量依赖性变化。PGI/PGII指标较干预前同样显著增加,但与 安慰剂组(D组)相比无显著差异。
研究表明,乳双歧杆菌BL-99干预,有助于提升PGI和PGI/PGII的指标水平,对于预防、改善慢性胃炎、萎缩性胃炎等胃部疾病具有良好的潜力。
胃泌素(G-17)是胃肠肽类激素之一,能够促进嗜铬细胞瘤释放组胺,刺激胃酸大量分泌,G-17值高于15会加剧胃炎发生,也是胃癌转变的前导性指标之一。反之,G-17<1则胃酸分泌过低,表现为胃动力不足、消化能力减弱、早饱、上腹饱胀等症状。经由乳双歧杆菌BL-99(E组、F组)干预2个月,期间检测G-17值,结果请参见图2与图3。结果显示,乳双歧杆菌BL-99(E组、F组)干预2个月后显著降低了G-17>15的人群G-17的数值,而对于胃酸分泌过低的人群,则经由BL-99干预一个月后,即显著的恢复到正常水平,在高剂量组(F组,500亿CFU)表现更为显著。
胃酸分泌失调是慢性胃炎的主要病因之一,使用奥美拉唑等质子泵抑制剂,抑制胃酸分泌也是临床对慢性胃炎最常见的治疗手段。然而越来越多的研究表明,长期使用质子泵抑制剂会导致胃肠道营养物质(维生素、矿物质等)吸收异常、胃肠道黏膜损伤、肾脏、心血管疾病发生风险增加、骨质代谢异常等。随着新技术的发展,质子泵抑制剂使用易引起肠道菌群的紊乱、与老年人阿尔兹海默症的发生也具有相关性。基于此,能够辅助减少质子泵抑制剂使用量、缓解副作用、甚至代替质子泵抑制剂的成分开发具有广泛的临床意义。
基于此,本发明发现了益生菌菌株乳双歧杆菌BL-99具有双向调节胃酸分泌量的效果。
胃癌前病变是一个病理性概念,包括肠上皮化生和异型增生,是从正常胃组织像胃癌转化过程的一个重要阶段。胃癌有很高的死亡率是因为转以后才被发现,所以“炎癌转化”也是临床关注的热点。近年来,随着胃癌前病变的不断研究,促炎因子在此过程中扮演的角色也不断地被定义。
白介素-6(IL-6)在机体中具有促炎、抗炎的双重作用,在炎症反应的刺激下,IL-6可以出现大量的表达,促使单核细胞、中性粒细胞释放,并在炎症部位大量聚集;TNF-α也是炎症过程中的一类重要介质,主要产生由于活化的单核细胞、巨噬细胞中,且且大量表达还会促使其他炎症因子的大量释放。随着胃粘膜炎症反应加剧,大量炎症因子释放并浸润胃部细胞,加速黏膜萎缩程度,对胃粘膜产生侵袭, 降低细胞内腺体分泌PG的能力,影响胃肠动力学。炎症因子IL-6、TNF-α和PGI,PGII,G-17等相互作用,共同影响胃部功能。
研究结果表明,乳双歧杆菌BL-99干预1个月后,可以显著降低炎症因子IL-6(图4)、TNF-α(图5)的表达,并且随着剂量增加,炎症因子的降低程度增加。表明乳双歧杆菌BL-99有助于降低炎症因子,对于胃炎相关的胃粘膜萎缩等症状具有缓解的潜力。
Claims (15)
- 一种乳双歧杆菌(Bifidobacterium lactis)在制备用于预防和/或改善胃炎的组合物中的应用,其中,所述乳双歧杆菌为保藏编号CGMCC No.15650的乳双歧杆菌。
- 根据权利要求1所述的应用,其中,所述预防和/或改善胃炎包括以下功效的一种或多种方面:改善功能性消化不良症状;提升胃蛋白酶原I(PG1)和/或PGI/PGII的指标水平;调节胃酸分泌量;和/或降低炎症因子IL-6和/或TNF-α的表达。
- 根据权利要求1所述的应用,其中,所述预防和/或改善胃炎包括:预防和/或改善慢性胃炎,和/或预防和/或改善胃炎相关的胃粘膜萎缩。
- 根据权利要求1所述的应用,其中,所述组合物包括食品组合物、饲料组合物或药品组合物。
- 根据权利要求1所述的应用,其中,所述乳双歧杆菌以固态或液态菌制剂的形式用于制备所述组合物。
- 根据权利要求1所述的应用,其中,所述乳双歧杆菌以活菌或灭活菌的形式用于制备所述组合物。
- 根据权利要求4所述的应用,其中,所述药品组合物还包括药学上可接受的辅料,所述辅料包括赋型剂、稀释剂、填充剂和/或吸收促进剂。
- 根据权利要求4-7中任一项所述的应用,其中,所述乳双歧杆菌的应用量为1.0×10 3CFU/天-1.0×10 12CFU/天,优选为1.0×10 7CFU/天-1.0×10 11CFU/天。
- 根据权利要求4-7中任一项所述的应用,其中,所述组合物为食品组合物。
- 根据权利要求9所述的应用,其中,所述食品为发酵乳制品、乳酪、含乳饮料、固体饮料、压片糖果、奶片、冰淇淋、乳粉。
- 一种用于预防和/或改善胃炎的组合物,其包括有效量的保藏编号CGMCC No.15650的乳双歧杆菌。
- 根据权利要求11所述的组合物,其中,所述组合物包括食品组合物、饲料组合物或药品组合物。
- 一种预防和/或改善胃炎的方法,该方法包括给予受试者有效量的保藏编号CGMCC No.15650的乳双歧杆菌。
- 根据权利要求13所述的方法,其中,所述预防和/或改善胃炎包括以下功效的一种或多种方面:改善功能性消化不良症状;提升胃蛋白酶原I(PG1)和/或PGI/PGII的指标水平;调节胃酸分泌量;和/或降低炎症因子IL-6和/或TNF-α的表达。
- 根据权利要求13或14所述的方法,其中,所述乳双歧杆菌给予受试者的量为1.0×10 3CFU/天-1.0×10 12CFU/天,优选为1.0×10 7CFU/天-1.0×10 11CFU/天。
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