Effects of placental periodontal pathogens on preeclampsia outcomes

doi:10.3877/cma.j.issn.1673-5250.2015.02.013

Objectlve

To investigate the expressions of serum C reactive protein (CRP), four common periodontal pathogens, porphyromonas gingivalis (Pg), actinobacillus actinomycetemcomitans (An), fusobacterium nucleatum (Fn), campylobacter rectus (Cr) and the alteration of NO formation in the placenta tissue of preeclampsia (PE) combined with periodontitis.

Methods

A total of 25 pregnant women who visited Guangdong Women and Children Hospital from January 2012 to October 2013 were included in this study (PE group). Meanwhile, another 27 healthy pregnant women were collected into control group. PE group and control group were both further divided into two sub-groups respectivley, PE group combined with periodontitis (PE-A sub-group, n=16), PE sub-group combined with no periodontitis (PE-B sub-group, n=9), control group combined with periodontitis (control-A sub-group, n=17), and control group combined with no periodontitis (control-B sub-group, n=10). There were no significant differences on age ratio, body weight, and body mass index (BMI) among four groups (P>0.05). The study protocol was approved by the Ethical Review Board of Investigation in Human Being of Guangdong Women and Children Hospital. Informed consent was obtained from the parent of each participates. CRP was tested by immunological transmission turbidimetry. Four periodontal pathogens in the placenta tissues were tested by PCR. The levels of endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) in placenta tissues were tested by Western blotting and Griess assay, respectively.

Results

①There were no significant differences on the incidence rate of periodontitis, PD, CAL and BOP between PE group and control group (P>0.05). ②There were significant differences on MSBP, MDBP, incidence rate of premature labor, and IUGR between PE-A sub-group and control-A sub-group, PE-A sub-group and control-B sub-group (MSBP: P<0.05; MDBP: P<0.05; incidence of premature delivery: P<0.05; IUGR: P<0.05). ③ The levels of MSBP, MDBP, incidence rate of premature labor, and IUGR between PE-B sub-group and control-A sub-group, PE-B sub-group and control-B sub-group (MSBP: t=50.5, 43.3; P<0.05; MDBP: t=13.3, 16.3; P<0.05; incidence of premature delivery: P<0.05; IUGR: P<0.05). ④ The incidence rate of fetal distress of PE-A sub-group was significant higher than that of control-B sub-group(P=0.018). ⑤ There were significant differences on gestational age, fetal birth weight, NO biosynthesis of placental tissues and placenta weight between PE-A sub-group and control-A sub-group, PE-A sub-group and control-B sub-group (gestational age: t=-7.0, -6.3; P<0.05; fetal birth weight: P<0.05; P<0.05; NO biosynthesis of placental tissues: t=-28.8, -42.1; P<0.05; placenta weight : P<0.05, P<0.001)). ⑥ There were significant differences on gestational age, fetal birth weight and placenta weight between PE-B sub-group and control-A sub-group, PE-B sub-group and control-B sub-group (gestational age: t=-5.9, -5.2, P<0.05; fetal birth weight: P<0.05; placenta weight: P<0.05, P<0.001 ). ⑦ The levels of serum CRP, incidence rates of Pg, Aa, and Cr in placental tissues of PE-A sub-groups were higher than those of control-B sub-group with significant differences (serum CRP: t=2.2, P=0.038; incidence rates of Pg, Aa, and Cr in placental tissues: χ²=5.097, P=0.024; χ²=7.659, P=0.006; P=0.044). ⑧ The b-actin/ eNOS value of PE-A sub-group was higher than that of PE-B sub-group, control-A sub-group and control-B sub-group, with significant differences (P<0.05). ⑨There were no significant differences on placental index among four groups (P>0.05).

Conclusions

Periodontal pathogens /byproducts may invade the placenta tissues and stimulate the inflammatory response against the activity of eNOS in placenta tissues. Then periodontal pathogens may lead to the exacerbation of PE adverse pregnancy outcomes.

Key words: Pre-eclampsia, Periodontitis, C-reactive protein, Nitric oxide synthase

表1 牙周细菌引物序列长度

Table 1 Sequences length of primer of periodental pathogens

牙周细菌	引物序列	扩增片段长度(bp)
Pg	正向引物5'-TGTAGATGACTGATGGTGAAAACC-3'	131
	反向引物5'-ACGTCATCCACACCTTCCTC-3'	?
Aa	正向引物5'-GCTAATACCGCCTAGAGTCGT-3'	235
	反向引物5'-ATTTCACACCTCACTTAAAGGT-3'	?
Fn	正向引物5'-GGCCACAAGGGGACTGAGACA-3'	185
?	反向引物5'-TTTAGCCGTCACTTCTTCTGTTGG-3'	?
Cr	正向引物5'-TTTCGGAGCGTAAACTCCTTT TC-3'	598
?	反向引物5'-TTTCTGCAAGCAGACACTCTT-3'	?

表1 牙周细菌引物序列长度

Table 1 Sequences length of primer of periodental pathogens

牙周细菌	引物序列	扩增片段长度(bp)
Pg	正向引物5'-TGTAGATGACTGATGGTGAAAACC-3'	131
	反向引物5'-ACGTCATCCACACCTTCCTC-3'	?
Aa	正向引物5'-GCTAATACCGCCTAGAGTCGT-3'	235
	反向引物5'-ATTTCACACCTCACTTAAAGGT-3'	?
Fn	正向引物5'-GGCCACAAGGGGACTGAGACA-3'	185
?	反向引物5'-TTTAGCCGTCACTTCTTCTGTTGG-3'	?
Cr	正向引物5'-TTTCGGAGCGTAAACTCCTTT TC-3'	598
?	反向引物5'-TTTCTGCAAGCAGACACTCTT-3'	?

表2 PE组和对照组孕妇牙周检查结果比较(±s)

Table 2 Comparison of periodontal examination between PE group and control group (±s)

组别	例数	牙周炎患病率例数(%)	PD(mm)	CAL(mm)	BOP例数(%)
PE组	25	16(64)	3.51±1.02	3.41±0.93	18(72.00)
对照组	27	10(37)	3.44±0.95	3.32±0.86	14(51.85)
t值/χ²值	?	3.775	0.980	0.158	2.23
P值	?	0.095	0.327	0.693	0.136

表2 PE组和对照组孕妇牙周检查结果比较(±s)

Table 2 Comparison of periodontal examination between PE group and control group (±s)

组别	例数	牙周炎患病率例数(%)	PD(mm)	CAL(mm)	BOP例数(%)
PE组	25	16(64)	3.51±1.02	3.41±0.93	18(72.00)
对照组	27	10(37)	3.44±0.95	3.32±0.86	14(51.85)
t值/χ²值	?	3.775	0.980	0.158	2.23
P值	?	0.095	0.327	0.693	0.136

表3 4组孕妇妊娠结局比较(±s)

Table 3 Comparison of pregnancy results among four groups (±s)

组别		例数	年龄(岁)	MSBP(mmHg)^a	MDBP (mmHg)^b	孕前BMI(kg/m²)	分娩孕龄(孕周)^c
PE组		?	?	?	?	?	?
?	PE-A亚组	16	28.0±4.4	156.2±21.7	95.6±16.8	23.4±3.3	32.9.±2.7
?	PE-B亚组	9	29.0±3.6	159.1±23.1	92.1± 9.2	23.1±2.6	33.9±2.5
对照组		?	?	?	?	?	?
?	对照-A亚组	10	25.0±3.6	108.1±13.1	78.1±9.2	21.3±4.2	39.9±0.8
?	对照-B亚组	17	26.0±7.2	116.1±12.4	75.7±7.8	21.6±4.4	39.1±0.9
t/χ²值		?	2.102	28.5	19.1	1.9	38.0
P值		?	0.112	0.000	0.000	0.139	0.000
组别		出生体质量(g)^d	胎儿宫内窘迫[例数(%)]^e	早产[例数(%)]^f	FGR[例数(%)]^g	生后1 min Apgar评分(分)	生后5 min Apgar评分(分)
PE组		?	?	?	?	?	?
?	PE-A亚组	2130±371	5(31)	13(81)	11(69)	8.6±0.7	9.8±0.4
?	PE-B亚组	2044±384	3 (8)	8(89)	5(56)	8.6±0.7	9.9±0.3
对照组		?	?	?	?	?	?
?	对照-A亚组	3184±308	1(10)	0 (0)	0 (0)	8.9±0.3	9.9±0.3
?	对照-B亚组	3284±358	0 (0)	0 (0)	0 (0)	9.0±0.0	10.0±0.0
t值/χ²值		44.7	10.1	48.4	32.0	7.7	1.1
P值		0.000	0.018	0.000	0.000	0.053	0.355

表3 4组孕妇妊娠结局比较(±s)

Table 3 Comparison of pregnancy results among four groups (±s)

组别		例数	年龄(岁)	MSBP(mmHg)^a	MDBP (mmHg)^b	孕前BMI(kg/m²)	分娩孕龄(孕周)^c
PE组		?	?	?	?	?	?
?	PE-A亚组	16	28.0±4.4	156.2±21.7	95.6±16.8	23.4±3.3	32.9.±2.7
?	PE-B亚组	9	29.0±3.6	159.1±23.1	92.1± 9.2	23.1±2.6	33.9±2.5
对照组		?	?	?	?	?	?
?	对照-A亚组	10	25.0±3.6	108.1±13.1	78.1±9.2	21.3±4.2	39.9±0.8
?	对照-B亚组	17	26.0±7.2	116.1±12.4	75.7±7.8	21.6±4.4	39.1±0.9
t/χ²值		?	2.102	28.5	19.1	1.9	38.0
P值		?	0.112	0.000	0.000	0.139	0.000
组别		出生体质量(g)^d	胎儿宫内窘迫[例数(%)]^e	早产[例数(%)]^f	FGR[例数(%)]^g	生后1 min Apgar评分(分)	生后5 min Apgar评分(分)
PE组		?	?	?	?	?	?
?	PE-A亚组	2130±371	5(31)	13(81)	11(69)	8.6±0.7	9.8±0.4
?	PE-B亚组	2044±384	3 (8)	8(89)	5(56)	8.6±0.7	9.9±0.3
对照组		?	?	?	?	?	?
?	对照-A亚组	3184±308	1(10)	0 (0)	0 (0)	8.9±0.3	9.9±0.3
?	对照-B亚组	3284±358	0 (0)	0 (0)	0 (0)	9.0±0.0	10.0±0.0
t值/χ²值		44.7	10.1	48.4	32.0	7.7	1.1
P值		0.000	0.018	0.000	0.000	0.053	0.355

表4 牙周菌在胎盘组织的分布及胎盘组织的NO含量情况

Table 4 The distribution of periodontal bacteria and the metabolism of NO in placenta

组别		例数	血清CRP^a	胎盘组织中检出牙周细菌^b				胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
组别		例数	血清CRP^a	Pg	Aa	Fn	Cr	胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
PE组		?	?	?	?	?	?	?	?	?	?
?	PE-A亚组	16	7.7±3.1	11(68.75)	9(56.00)	7(43.75)	4(25.00)	16.1±2.6	35.5±16.7	0.2±0.1	467.9±104.4
?	PE-B亚组	9	7.1±2.2	3(33.33)	3(33.33)	2(22.22)	1(11.11)	12.7±3.5	49.7±15.0	0.2±0.0	441.0± 97.3
对照组		?	?	?	?	?	?	?	?	?	?
?	对照-A亚组	10	5.8±2.3	4(40.00)	3(30.00)	2(20.00)	0(0.00)	10.8±2.6	64.3±23.6	0.2±0.0	593.4±119.9
?	对照-B亚组	17	5.9±1.9	4(23.50)	1 (5.88)	2(11.76)	0(0.00)	7.7±2.1	77.6±16.8	0.2±0.0	603.5± 83.7
t值/χ²值		?	2.0	10.1	10.98	4.695	8.647	28.3	17.2	2.7	8.8
P值		?	0.126	0.018	0.012	0.196	0.034	0.000	0.000	0.057	0.000

表4 牙周菌在胎盘组织的分布及胎盘组织的NO含量情况

Table 4 The distribution of periodontal bacteria and the metabolism of NO in placenta

组别		例数	血清CRP^a	胎盘组织中检出牙周细菌^b				胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
组别		例数	血清CRP^a	Pg	Aa	Fn	Cr	胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
PE组		?	?	?	?	?	?	?	?	?	?
?	PE-A亚组	16	7.7±3.1	11(68.75)	9(56.00)	7(43.75)	4(25.00)	16.1±2.6	35.5±16.7	0.2±0.1	467.9±104.4
?	PE-B亚组	9	7.1±2.2	3(33.33)	3(33.33)	2(22.22)	1(11.11)	12.7±3.5	49.7±15.0	0.2±0.0	441.0± 97.3
对照组		?	?	?	?	?	?	?	?	?	?
?	对照-A亚组	10	5.8±2.3	4(40.00)	3(30.00)	2(20.00)	0(0.00)	10.8±2.6	64.3±23.6	0.2±0.0	593.4±119.9
?	对照-B亚组	17	5.9±1.9	4(23.50)	1 (5.88)	2(11.76)	0(0.00)	7.7±2.1	77.6±16.8	0.2±0.0	603.5± 83.7
t值/χ²值		?	2.0	10.1	10.98	4.695	8.647	28.3	17.2	2.7	8.8
P值		?	0.126	0.018	0.012	0.196	0.034	0.000	0.000	0.057	0.000

[1]	Matevosyan NR. Periodontal disease and perinatal outcomes [J]. Arch Gynecol Obstet, 2011, 283(4): 675–686.
[2]	Kumar A, Basra M, Begum N, et al. Association of maternal periodontal health with adverse pregnancy outcome [J]. J Obstet Gynaecol Res, 2013, 39(1): 40–45.
[3]	Madianos PN, Bobetsis YA, Offenbacher S. Adverse pregnancy outcomes (APOs) and periodontal disease: pathogenic mechanisms [J]. J Periodontol, 2013, 84(4 Suppl): S170–S180.
[4]	Sayar F, Hoseini MS, Abbaspour S. Effect of periodontal disease on preeclampsia [J]. Iran J Public Health, 2011, 40(3): 122–127.
[5]	Ha JE, Oh KJ, Yang HJ, et al. Oral health behaviors, periodontal disease, and pathogens in preeclampsia: a case-control study in Korea [J]. J Periodontol, 2011, 82(12): 1685–1692.
[6]	Sun W, Wu J, Lin L, et al. Porphyromonas gingivalis stimulates the release of nitric oxide by inducing expression of inducible nitric oxide synthases and inhibiting endothelial nitric oxide synthases [J]. J Periodontal Res, 2010, 45(3): 381–388.
[7]	Barak S, Oettinger-Barak O, Machtei EE, et al. Evidence of periopathogenic microorganisms in placentas of women with preeclampsia [J]. J Periodontol, 2007, 78(4): 670–676.
[8]	Kumar A, Basra M, Begum N, et al. Association of maternal periodontal health with adverse pregnancy outcome [J]. J Obstet Gynaecol Res, 2013, 39(1): 40–45.
[9]	Rylev M, KilianM. Prevalence and distribution of principal periodontal pathogens worldwide [J]. J Clin Periodontol, 2008, 35(8 Suppl): 346–361.
[10]	Wang Y, Sugita N, Kikuchi A, et al. FcγRIIB-nt645＋25A/G gene polymorphism and periodontitis in Japanese women with preeclampsia [J]. Int J Immunogenet, 2012, 39(6): 492–500.
[11]	Li Y, Shibata Y, Zhang L, et al. Periodontal pathogen Aggregatibacter actinomycetemcomitans LPS induces mitochondria-dependent-apoptosis in human placental trophoblasts [J]. Placenta, 2011, 32(1): 11–19.
[12]	Liu H, Redline RW, Han YW. Fusobacterium nucleatum induces fetal death in mice via stimulation of TLR4-mediated placental inflammatory response [J]. J Immunol, 2007, 179(4): 2501–2508.
[13]	Offenbacher S, Riché EL, Barros SP, et al. Effects of maternal Campylobacter rectus infection on murine placenta, fetal and neonatal survival, and brain development [J]. J Periodontol, 2005, 76(11 Suppl): 2133–2143.
[14]	Horton AL, Boggess KA, Moss KL, et al. Periodontal disease early in pregnancy is associated with maternal systemic inflammation among African American women [J]. J Periodontol, 2008, 79(7): 1127–1132.
[15]	Chaparro A, Sanz A, Quintero A, et al. Increased inflammatory biomarkers in early pregnancy is associated with the development of pre-eclampsia inpatients with periodontitis: a case control study [J]. J Periodontal Res, 2013, 48(3): 302–307.
[16]	Rylev M, Kilian M. Prevalence and distribution of principal periodontal pathogens worldwide [J]. J Clin Periodontol, 2008, 35(8 Suppl): 346–361.
[17]	Mineo C, Gormley AK, Yuhanna IS, et al. FcgammaRIIB mediates C-reactive protein inhibition of endothelial NO synthase[J]. Circ Res, 2005, 97(11): 1124–1131.
[18]	Kulandavelu S, Whiteley KJ, Qu D, et al. Endothelial nitric oxide synthase deficiency reduces uterine blood flow, spiral artery elongation, and placentaloxygenation in pregnant mice [J]. Hypertension, 2012, 60(1): 231–238.
[19]	Kulandavelu S, Whiteley KJ, Bainbridge SA, et al. Endothelial NO synthase augments fetoplacental blood flow, placental vascularization, and fetal growth in mice [J]. Hypertension, 2013, 61(1): 259–266.
[20]	Fernández CL, Carbajo RM, Muoz RM. Prolonged inhibition of nitric oxide synthesis in pregnant rats: effects on blood pressure, fetal growth and litter size [J]. Arch Gynecol Obstet, 2005, 271(3): 243–248.

[1]	Minghao Lu, Hong Li, Benxiang Hou. Research progress of periapical biofilm removal methods[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(04): 257-261.
[2]	Ka Teng Chan, Meijun Du, Bing Shi, Hanyao Huang. The impact of maternal systemic diseases on the incidence of cleft lip and palate in newborns[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(04): 262-268.
[3]	Hongkun Sun, Hong Ai, Zheng Chen. Advances in endoplasmic reticulum stress-mediated bone remodeling imbalance in periodontitis[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(04): 211-218.
[4]	Zekai Liao, Ailin Liang, Qimei Gong. Research progress on programmed cell death in periapical periodontitis[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(03): 150-155.
[5]	Yeqing Wang, Litong Li, Weixu Li, Meng Cao. The causal association between periodontitis and diabetic retinopathy: A bidirectional two-sample mendelian randomization analysis[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(03): 160-168.
[6]	Yuanjie Luo, Jingmei Yang, Shu Meng, Yibo Ao, Daonan Shen. Progress of research on quercetin against oral diseases[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(02): 117-122.
[7]	Lei Dong, Xiuzhen Li, Linzi Han, Fang Yang. Clinical observation of pulp revascularization in the treatment of young permanent molars with chronic apical periodontitis[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2024, 18(01): 38-42.
[8]	Ding Ding, Yunchuan Yang, Xiang Ma, Zhongzheng Ma, Junyi Huo, Lei Zhou. Evaluation of the prognostic value of preoperative C-reactive protein-albumin-lymphocyte ratio in patients with hepatocellular carcinoma[J]. Chinese Archives of General Surgery(Electronic Edition), 2024, 18(04): 261-265.
[9]	Ding Wei, Yanyan Qiao, Xing Gu, Yan Zhang, Yanyan Li, Weisheng Qian, Lei Pan, Yongheng Gao, Faguang Jin. Risk factors of poor prognosis after extracorporeal membrane oxygenation in acute respiratory distress syndrome patients[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(03): 363-367.
[10]	Rui Liu, Le Li, Jinming Chen, Xin Li. Meta-analysis of the correlation between the severity of acute cholangitis and serum markers[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2024, 13(02): 176-181.
[11]	Sihan Liu, Jing Zhang. Relationship of 25-hydroxyvitamin D3 and the ratio of serum iron to C-reactive protein with renal function impairment in the elderly[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2024, 13(05): 268-272.
[12]	Yin Zhang, Guoqiang Li. Treatment of septic shock with methylene blue[J]. Chinese Journal of Critical Care & Intensive Care Medicine(Electronic Edition), 2024, 10(02): 143-147.
[13]	Wenkai Sun, Qing Shen, Li Hang, Yingchun Zhang. Relationship of FAR, NAR, CRP with the evaluation and prognosis of ulcerative colitis patients[J]. Chinese Journal of Digestion and Medical Imageology(Electronic Edition), 2024, 14(05): 426-431.
[14]	Qinqin Zhu, Juanjuan Ci, Lu Cui, Hairong Xu, Yuxin Li, Yanbo Ding. Value of coagulation function index, blood lipid level, C-reactive protein and neutrophil-lymphocyte ratio in the activity evaluation and clinical diagnosis of Crohn's disease[J]. Chinese Journal of Digestion and Medical Imageology(Electronic Edition), 2024, 14(01): 35-40.
[15]	Chunying Ou, Xiaobin Li, Jing Guo, Ke Xu, Meng Wang, Xiaolei An. Correlation of hs-CRP, Lp-PLA2, and S100β with vascular cognitive impairment in patients with ischemic small vascular disease[J]. Chinese Journal of Cerebrovascular Diseases(Electronic Edition), 2024, 18(03): 265-269.

Viewed

Full text

Abstract

Please choose a citation manager

Content to export