Effect of Liangfang Wenjing Decoction on expression of key glycolytic enzymes in uterus and ovaries of rats with coagulating cold and blood stasis syndrome / 中国中药杂志

This study aimed to investigate the relationship between coagulating cold and blood stasis syndrome and glycolysis, and observe the intervention effect of Liangfang Wenjing Decoction(LFWJD) on the expression of key glycolytic enzymes in the uterus and ovaries of rats with coagulating cold and blood stasis. The rat model of coagulating cold and blood stasis syndrome was established by ice-water bath. After modeling, the quantitative scoring of symptoms were performed, and according to the scoring results, the rats were randomly divided into a model group and LFWJD low-, medium-and high-dose groups(4.7, 9.4, 18.8 g·kg~(-1)·d~(-1)), with 10 in each group. Another 10 rats were selected as the blank group. After 4 weeks of continuous administration by gavage, the quantitative scoring of symptoms was repeated. Laser speckle flowgraphy was used to detect the changes of microcirculation in the ears and uterus of rats in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological morphology of uterus and ovaries of rats in each group. The mRNA and protein expressions of pyruvate dehydrogenase kinase 1(PDK1), hexokinase 2(HK2) and lactate dehydrogenase A(LDHA) in the uterus and ovaries of rats were examined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot, respectively. The rats in the model group showed signs of coagulating cold and blood stasis syndrome, such as curl-up, less movement, thickened veins under the tongue, and reduced blood perfusion in the microcirculation of the ears and uterus, and HE staining revealed a thinning of the endometrium with disorganized arrangement of epithelial cells and a decrease in the number of ovarian follicles. Compared with the model group, the treatment groups had alleviated coagulating cold and blood stasis, which was manifested as red tongue, reduced nail swelling, no blood stasis at the tail end as well as increased blood perfusion of the microcirculation in the ears and uterus(P<0.05 or P<0.01). Among the groups, the LFWJD medium-and high-dose groups had the most significant improvement in coagulating cold and blood stasis, with neatly arranged columnar epithelial cells in uterus, and the number of ovarian follicles was higher than that in the model group, especially mature follicles. The mRNA and protein expressions of PDK1, HK2, LDHA in uterus and ovaries were up-regulated in the model group(P<0.05 or P<0.01), while down-regulated in LFWJD medium-and high-dose groups(P<0.05 or P<0.01). The LFWJD low-dose group presented a decrease in the mRNA expressions of PDK1, HK2 and LDHA in uterus and ovaries as well as in the protein expressions of HK2 and LDHA in uterus and HK2 and PDK1 in ovaries(P<0.05 or P<0.01). The therapeutic mechanism of LFWJD against coagulating cold and blood stasis syndrome is related to the down-regulation of key glycolytic enzymes PDK1, HK2 and LDHA, and the inhibition of glycolytic activities in uterus and ovaries.

Modified Qiaohetang and Xiehuangsan in Treatment of Acne Due to Dampness-heat Accumulation / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of modified Qiaohetang and Xiehuangsan in the treatment of acne due to dampness-heat accumulation and its influence on the levels of inflammatory factors and sex hormones. Method:One hundred and sixty-eight eligible patients were divided into an observation group （84 cases） and a control group （84 cases） according to the random number table. Adapalene gel was applied externally in both groups， one time per day. In the control group， Jinhua Xiaocuo pills was taken orally， 4 g per time， three times a day. In the observation group， the modified Qiaohetang and Xiehuangsan was provided for oral administration， one bag per day. The treatment lasted for eight weeks. The Global Acne Grading System （GAGS） score， skin lesion count， dampness-heat accumulation syndrome score， and Dermatology Life Quality Index （DLQI） score were recorded before and after treatment， followed by the detection of interleukin-8 （IL-8）， IL-10， IL-17， interferon-<italic>γ</italic> （IFN-<italic>γ</italic>）， free testosterone （FT）， estradiol （E₂） and sex hormone binding globulin （SHBG） before and after treatment as well as the safety evaluation. Result:The GAGS， dampness-heat accumulation syndrome， and DLQI scores of the observation group were lower than those of the control group （<italic>P</italic><0.01）. The counts of inflammatory skin lesions （papule and pustule）， non-inflammatory skin lesions， and total skin lesions in the observation group declined in contrast to those in the control group （<italic>P</italic><0.01）. The IL-8， IL-17， IFN-<italic>γ</italic> and FT levels of the observation group were decreased as compared with those of the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）， while the IL-10， E₂， and SHBG levels were increased （<italic>P</italic><0.01）. The overall response rate in clinical symptom alleviation of the observation group was 93.67%（74/79）， which was higher than 81.82%（63/77） of the control group （<italic>χ</italic>²=5.121， <italic>P</italic><0.05）. The overall response rate in dampness-heat accumulation syndrome relief of the observation group was 92.41% （73/79）， still higher than 79.22% （61/77） of the control group （<italic>χ</italic>²=5.595， <italic>P</italic><0.05）. No adverse reactions occurred after the oral administration of Chinese medicinal preparations. Conclusion:The modified Qiaohetang and Xiehuangsan combined with adapalene gel can reduce the skin lesion count and severity， relieve both clinical symptoms and dampness-heat accumulation syndrome， regulate the inflammatory response and sex hormones， and improve the quality of life of patients with acne of dampness-heat accumulation syndrome without inducing side effects.

Mechanism of Wenjingtang on Improving Endometriosis in Rats Based on Regulating HIF-1α Expression / 中国实验方剂学杂志

Objective:To observe the effect of Wenjingtang on the expression of hypoxia inducible factor-1α （HIF-1α） and ultrastructure of mitochondria in rats with endometriosis （EMs）， and to explore the mechanism of Wenjingtang in the treatment of EMs. Method:The EMs model rat was established by autogenous intimal transplantation， and the volume of ectopic lesions was measured by small animal ultrasound imaging system. According to the volume of ectopic lesions， rats successfully modeled were randomly divided into model group， low， middle ang high dose Wenjingtang group （4.85， 9.7， 19.4 g·kg-1） and gestrinone group （0.25 mg·kg-1）， 10 in each group， another 10 rats were recruited in a sham operation group. After 6 weeks of drug treatment， the volume of ectopic lesions was measured by ultrasound imaging system and caliper， the morphology of ectopic endometrium was observed by hematoxylin-eosin （HE） staining， the levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α） and transforming growth factor-β1 （TGF-β1） in peritoneal fluid were measured by enzyme linked immunosorbent assay （ELISA）， the mRNA and protein expression of HIF-1α in eutopic or ectopic endometrial tissues were detected by Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， and the ultrastructure of mitochondria in ectopic endometrium was observed by transmission electron microscope （TEM）. Result:Compared with sham operation group， ectopic lesions were found in model group， showing vesicular structure， and the typical endometrial tissue morphology was found in the lesions. The contents of IL-1β， TNF-α， TGF-β1 and the expression of HIF-1α mRNA and protein were significantly increased than those in the sham operation group （P<0.05，P<0.01）. TEM showed that the number of mitochondria in the cytoplasm of ectopic endometrium increased and the structure was intact. Compared with model group， the volume of ectopic focus was significantly decreased in all Wenjingtang groups （P<0.01）， and the results of ultrasonic examination were basically consistent with those of caliper measurement， HE staining showed that the columnar epithelial cells of ectopic endometrium were damaged or exfoliated and the interstitial cells were loose， the content of TNF-α in each dose group of Wenjingtang was significantly decreased， the content of IL-1β and TGF-β1 in the medium dose and high dose groups of Wenjingtang was significantly decreased （P<0.05，P<0.01）， the expression of HIF-1α mRNA and protein were significantly decreased in all Wenjingtang groups （P<0.05，P<0.01）. The mitochondria of ectopic endometrium were obviously swollen， the crest was broken or even disappeared， some of the mitochondria were vacuolar degeneration and the outer membrane was ruptured. Conclusion:Wenjingtang has a good therapeutic effect on experimental EMs in rats， and the mechanism is related to reducing the expression of HIF-1α， improving hypoxia in ectopic lesions and inducing mitochondrial damage in ectopic endometrium.

Establishment of gynecological asthenia cold syndrome model / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To establish the model of gynecological asthenia cold syndrome by simulating the etiology of gynecological asthenia cold syndrome depending on 'pathogenic cold impairing yang" in Chinese medicine theory.</p><p><b>METHODS</b>The female SD rats were randomly divided into the model group and the normal group by randomized digital table, 20 in each group. Rats in the model group were placed in 0 degrees C - 1 degree C ice water, and the ice water was placed in 4 degrees C refrigerator, twice daily, 20 min each time, for a total of 30 days. The body temperature was determined and the changes of the estrous cycle were observed every day. When the body temperature decreased (with statistical difference from those of the normal group), and vaginal smears showed disordered estrous cycle, the model was successfully established. Rats were sacrificed during the diestrus period, the correlative indices including reproductive endocrine hormones in blood [follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol ( E2 ), testosterone (T), and progestone (P)], the thyroid function [triiodothyronine (T3), tetraiodothyronine (T4), thyrotropic-stimulating hormone (TSH)], the adrenal function [plasma adrenocorticotropic hormone (ACTH) and cortisol (Cor)], the cellular immune function [serum interleukin-2 (IL-2)], and energy tests [including plasma cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), the ratio of cAMP/cGMP, lactate dehydrogenase(LDH)], an the index of thymus, spleen, uterus, and ovary were detected.</p><p><b>RESULTS</b>Compared with the normal group, rats in the model group appeared chill, dim-color hair, purple and dark ears and claw. They were depressed, scrunched, quiet, and clumsy. They liked to stay together. Their water intake and appetite were reduced, body weight lost, body temperature significantly lowered. They passed loose stool. Their estrous cycle and diestrus were prolonged. Their plasma cGMP content, cAMP/cGMP ratio, LDH, serum IL-2 content, E2, P, T, T3, LH, TSH were significantly lowered to some extent. Their thymus index and the ovary index significantly decreased, showing significant difference (P < 0.05).</p><p><b>CONCLUSION</b>The rat model of gynecological asthenia cold syndrome prepared by extending the frozen time at refrigerator and ice water immersion was in line with clinical features of gynecological asthenia cold syndrome.</p>