白头翁皂苷B4对临床型奶牛乳房炎疗效和血清炎性因子、免疫因子的影响

doi:10.3969/j.issn.1004-1524.2021.07.04

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (7): 1184-1191.DOI: 10.3969/j.issn.1004-1524.2021.07.04

白头翁皂苷B4对临床型奶牛乳房炎疗效和血清炎性因子、免疫因子的影响

沈留红¹(), 钱柏霖¹, 尤留超¹, 张钺¹, 莘余¹, 吕尚揆¹, 肖劲邦¹, 余树民¹, 苏柘僮², 董可³, 杨世林⁴, 冯育林⁴^,^*(), 曹随忠¹^,^*()

1.四川农业大学动物医学院,动物疫病与人类健康四川省重点实验室/奶牛疾病研究中心,四川成都 611130
2.广西英路维特药物有限公司,广西荔浦 541000
3.四川育强本草生物技术有限公司,四川成都 611130
4.江西中医药大学中药固体制剂制造技术国家工程研究中心,江西南昌 330006

收稿日期:2020-12-10 出版日期:2021-07-25 发布日期:2021-08-06
通讯作者: 冯育林,曹随忠
作者简介:曹随忠,E-mail: suizhongcao@126.com
*冯育林,E-mail: fengyulin2003@126.com;
沈留红(1979—),男,江苏如皋人,博士,副教授,研究方向为反刍动物疾病与繁殖新技术。E-mail: shenlh@sicau.edu.cn
基金资助:
国家重点研发计划(2018YFD0500900);四川省科技计划(2019YJ0650);江苏农牧科技职业学院产业发展关键技术创新项目(NSF2021ZR01);四川农业大学学科建设双支计划(03572174);四川农业大学本科科研兴趣培养计划(KY2021140)

Effect of Pulsatilla saponin B4 on treatment efficiency and serum inflammatory and immune factors of dairy cows with clinical mastitis

SHEN Liuhong¹(), QIAN Bolin¹, YOU Liuchao¹, ZHANG Yue¹, SHEN Yu¹, LYU Shangkui¹, XIAO Jinbang¹, YU Shumin¹, SU Zhetong², DONG Ke³, YANG Shilin⁴, FENG Yulin⁴^,^*(), CAO Suizhong¹^,^*()

1. Key Laboratory of Animal Disease and Human Health of Sichuan Province/The Medical Research Center for Cow Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2. Guangxi Innovate Medical Technology Co., Ltd., Lipu 541000, China
3. Sichuan Yuqiang Materia Medica Biotechnology Co., Ltd., Chengdu 611130, China
4. National Pharmaceutical Engineering Center for Solid Preparation in Chinese Herbal Medicine, Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China

Received:2020-12-10 Online:2021-07-25 Published:2021-08-06
Contact: FENG Yulin,CAO Suizhong

摘要/Abstract

摘要：

为观察不同剂量白头翁皂苷B4对临床型奶牛乳房炎的疗效,及对血清炎性因子和免疫因子的影响,将36头患临床型乳房炎的奶牛随机分为低、中、高剂量3个试验组,同时选择12头健康奶牛作为对照组。试验组每日分别于颈部肌肉注射白头翁皂苷B4 0.025、0.05、0.1 mL·kg^-1,连续注射6 d,对照组不注射任何药物。试验期间,观察试验组各奶牛的有效率和治愈率,同时采集对照组任意一天,及试验组1、3、5、7 d静脉血进行炎性因子和免疫因子检测,分别采集试验组1、7、12 d患病乳区奶样进行乳汁体细胞数(SCC)计数和病原微生物检测,整个试验观察至12 d,统计临床疗效。临床疗效试验结果表明,白头翁皂苷B4低、中、高3个剂量组的临床治愈率和总有效率分别为75.00%、75.00%、91.67%和100%、100%、100%,平均治愈时间分别为6.33、6.11、6.91 d。白头翁皂苷B4可显著(P<0.05)降低临床型奶牛乳房炎SCC。治疗前,36份奶样有34份检测出病原微生物,检出率为94.44%,主要为支原体、球菌和大肠埃希菌等;治疗12 d后,病原微生物总转阴率为52.94%,其中,杆菌、产色葡萄球菌、溶血葡萄球菌和大肠埃希菌的转阴率均达100%,支原体、金黄色葡萄球菌、表皮葡萄球菌和球菌的转阴率分别为69.23%、60.00%、50.00%和33.33%。白头翁皂苷B4可显著(P<0.05)促使临床型奶牛乳房炎血清炎性因子CRP、SAA、HP、IL-1α、IL-1β、IL-2、IL-6、IL-8、TNF-α和免疫因子IgA、IgD、IgE、IgG、IgM恢复到健康水平。

关键词: 白头翁皂苷B4, 临床型奶牛乳房炎, 疗效, 炎性因子, 免疫因子

Abstract:

To investigate the effects of Pulsatilla saponin B4 on therapeutic effect, serum inflammatory and immune factors in dairy cows with clinical mastitis, 12 healthy dairy cows were selected as the control group, and 36 clinical mastitis cows were selected as the test group and were randomly divided into low, middle and high dose group. Cows in test group were administered with Pulsatilla saponin B4 injection (with doses of 0.025, 0.05, 0.1 mL·kg^-1, respectively) through cervical intramuscular injection once a day for 6 days, whereas no medicine was administered in the control group. The effective and cure rates of cows in the test group were recorded, venous blood samples were collected (any day in the control group and the 1^st, 3^rd, 5^th, 7^th day in the test group) to measure serum inflammatory and immune factors, as well as milk in the infected quarters on the 1^st, 7^th, 12^th day in the test group to conduct somatic cells count (SCC) and pathogenic microorganism isolation and identification during this experiment. Cows were observed till the 12^th day and the therapeutic efficiency was recorded. It was shown that the clinical cure rate and effective rates of low, middle and high dose group were 75.00%, 75.00%, 91.67% and 100%, 100%, 100%, respectively, and the average cure time was 6.33, 6.11, 6.91 d, respectively. The Pulsatilla saponin B4 could significantly (P<0.05) attenuate SCC. Before treatment, 34 out of 36 milk samples were positive with pathogenic microorganism, indicating a detection rate of 94.44%, which mainly consisted of mycoplasma, cocci and Escherichia coli. After treatment, the total negative rate was 52.94%, among which, the negative rate of Bacilli, Staphylococcus chromogenes, Staphylococcus haemolyticus and Escherichia coli were 100%, while the negative rates of mycoplasma, Staphylococcus aureus, Staphylococcus epidermidis and coccus were 69.23%, 60.00%, 50.00% and 33.33%, respectively. Besides, Pulsatilla saponin B4 could significantly (P<0.05) promote the recovery of inflammatory factors CRP, SAA, HP, IL-1α, IL-1β, IL-2, IL-6, IL-8, TNF-α and immune factors IgA, IgD, IgE, IgG, IgM to normal levels.

Key words: Pulsatilla saponins B4, clinical mastitis, therapeutic effect, inflammatory factors, immune factors

中图分类号:

S854.5

沈留红, 钱柏霖, 尤留超, 张钺, 莘余, 吕尚揆, 肖劲邦, 余树民, 苏柘僮, 董可, 杨世林, 冯育林, 曹随忠. 白头翁皂苷B4对临床型奶牛乳房炎疗效和血清炎性因子、免疫因子的影响[J]. 浙江农业学报, 2021, 33(7): 1184-1191.

SHEN Liuhong, QIAN Bolin, YOU Liuchao, ZHANG Yue, SHEN Yu, LYU Shangkui, XIAO Jinbang, YU Shumin, SU Zhetong, DONG Ke, YANG Shilin, FENG Yulin, CAO Suizhong. Effect of Pulsatilla saponin B4 on treatment efficiency and serum inflammatory and immune factors of dairy cows with clinical mastitis[J]. Acta Agriculturae Zhejiangensis, 2021, 33(7): 1184-1191.

图/表 5

表1 不同剂量白头翁皂苷B4对临床型奶牛乳房炎乳汁体细胞数的影响

Table 1 Influence of different dosage of Pulsatilla saponin B4 on somatic cell counts in milk of dairy cows with clinical mastitis

组别 Group	第1天 1^st day	第7天 7^th day	第12天 12^th day
B	255.8±38. $9 a a$	90.5±55. $8 ab b$	47.5±35. $6 ab b$
C	244.6±42. $0 a a$	65.8±51. $5 b b$	38.1±23. $0 b b$
D	310.6±132. $4 a a$	182.3±151. $7 a b$	48.8±58. $9 a c$

表1 不同剂量白头翁皂苷B4对临床型奶牛乳房炎乳汁体细胞数的影响

Table 1 Influence of different dosage of Pulsatilla saponin B4 on somatic cell counts in milk of dairy cows with clinical mastitis

组别 Group	第1天 1^st day	第7天 7^th day	第12天 12^th day
B	255.8±38. $9 a a$	90.5±55. $8 ab b$	47.5±35. $6 ab b$
C	244.6±42. $0 a a$	65.8±51. $5 b b$	38.1±23. $0 b b$
D	310.6±132. $4 a a$	182.3±151. $7 a b$	48.8±58. $9 a c$

表2 不同剂量白头翁皂苷B4对临床型奶牛乳房炎治愈时间和治愈率的影响

Table 2 Influence of different dosage of Pulsatilla saponin B4 on cure time and cure rate of dairy cows with clinical mastitis

指标 Index	B组 Group B	C组 Group C	D组 Group D
4 d内治愈数	0	1	1
Cured number within 4 days
5 d内治愈数	2	2	2
Cured number within 5 days
6 d内治愈数	4	6	4
Cured number within 6 days
7 d内治愈数	9	8	9
Cured number within 7 days
治愈数 Cured number	9	9	11
有效数Effective number	3	3	1
无效数 Recurrence number	0	0	0
治愈率Cure rate/%	75.00	75.00	91.67
总有效率 Total effective rate/%	100	100	100
平均治愈时间	6.33	6.11	6.91
Average cure time/d

表3 不同剂量白头翁皂苷B4对临床型奶牛乳房炎病原微生物的影响

Table 3 Influence of different dosage of Pulsatilla saponins B4 on pathogenic microorganism of dairy cows with clinical mastitis

病原微生物Pathogenic microorganism	治疗前 Before treatment	治疗后 After treatment	转阴率 Negative rate%
表皮葡萄球菌 Staphylococcus epidermidis	2	1	50.00
杆菌Bacilli	1	0	100
阿尔莱特葡萄球菌 Staphylococcus arlettae	1	1	0
支原体Mycoplasma	13	4	69.23
球菌Coccus	6	4	33.33
产色葡萄球菌 Staphylococcus chromogenes	1	0	100
溶血葡萄球菌 Staphylococcus haemolyticus	1	0	100
腐生葡萄球菌Staphylococcus saprophyticus	1	1	0
大肠埃希菌 Escherichia coli	5	0	100
金黄色葡萄球菌 Staphylococcus aureus	2	2	0
葡萄球菌 Staphylococcus	3	3	0

表4 白头翁皂苷B4对临床型奶牛乳房炎血清炎性因子的影响

Table 4 Influence of Pulsatilla saponin B4 on serum inflammatory factor index of dairy cows with clinical mastitis

指标 Index	对照组 Control group	试验组 Test group
指标 Index	对照组 Control group	第1天 1^st day	第3天 3^rd day	第5天 5^th day	第7天 7^th day
CRP/(mg·mL^-1)	6.55±1.68 b	8.25±2.50 a	7.02±2.08 ab	6.66±1.05 b	6.47±1.93 b
SAA/(μg·mL^-1)	6.95±1.01 c	10.86±3.20 a	9.40±2.63 ab	8.85±2.31 bc	8.71±2.60 bc
HP/(ng·mL^-1)	167.37±36.40 b	236.34±55.99 a	178.46±45.14 b	168.68±35.05 b	160.01±52.80 b
LTB4/(ng·mL^-1)	11.15±2.69 b	14.71±3.82 a	14.49±4.77 a	14.27±4.68 a	14.13±4.78 a
PGE2/(pg·mL^-1)	253.70±52.08 c	351.80±56.39 a	331.47±82.63 ab	307.83±66.35 b	299.62±51.36 b
IL-1α/(pg·mL^-1)	175.90±47.49 bc	228.47±69.62 a	210.98±63.04 ab	181.65±60.07 bc	163.23±47.55 c
IL-1β/(pg·mL^-1)	276.71±81.61 b	358.78±113.73 a	331.88±99.60 ab	325.92±98.89 ab	295.64±98.06 ab
IL-2/(pg·mL^-1)	449.65±95.09 b	606.38±130.78 a	500.38±151.23 b	470.21±145.64 b	415.76±121.10 b
IL-6/(pg·mL^-1)	97.67±22.63 c	155.00±39.96 a	134.52±38.21 ab	124.65±42.53 b	117.07±36.88 bc
IL-8/(pg·mL^-1)	108.61±38.62 b	165.45±53.22 a	153.94±50.83 a	152.60±54.40 a	139.76±47.86 ab
TNF-α/(pg·mL^-1)	122.65±33.50 c	174.65±49.07 a	151.22±41.09 ab	143.77±35.69 bc	133.20±46.61 bc

表5 白头翁皂苷B4对临床型奶牛乳房炎血清免疫因子的影响

Table 5 Influence of Pulsatilla saponin B4 on serum immune factor index of dairy cows with clinical mastitis μg·mL-1

指标 Index	对照组 Control group	试验组 Test group
指标 Index	对照组 Control group	第1天 1^st day	第3天 3^rd day	第5天 5^th day	第7天 7^th day
IgA	147.58±30.60 c	239.50±38.16 a	197.57±36.29 b	196.53±40.99 b	178.71±38.18 bc
IgD	12.95±2.73 b	17.75±5.42 a	16.13±3.93 ab	15.40±4.66 ab	14.49±4.69 b
IgE	879.69±170.18 c	1 291.81±355.69 a	1 132.70±311.68 ab	1 129.13±311.64 ab	1 006.83±417.28 bc
IgG	182.37±33.29 c	275.88±74.13 a	234.38±65.60 b	229.49±54.44 b	221.81±54.44 bc
IgM	111.77±31.35 c	152.14±43.25 a	141.73±45.04 ab	137.82±34.90 ab	126.00±32.96 bc

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白头翁皂苷B4对临床型奶牛乳房炎疗效和血清炎性因子、免疫因子的影响

Effect of Pulsatilla saponin B4 on treatment efficiency and serum inflammatory and immune factors of dairy cows with clinical mastitis

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