ABSTRACT
Objective:To study the regulative effect of Rhubarb dispensing granule on autophagy and gastrointestinal motility of Cajal interstitial cells in rats with chronic transit constipation (STC).Methods:A total of 75 rats were randomly divided into control group, model group, low, medium and high dose groups with 15 rats in each group. Except for the control group, the STC rat models were established by intragastric administration of compound diphenoxylate suspension. Rats in low, medium and high dose groups were given Rhubarb dispensing granule of 1, 2 and 4 g/kg by gavage respectively, while rats in control group and model group were given normal saline with the same volume by gavage, once a day for 14 consecutive days. Fecal moisture content and intestinal propulsion rate were measured. The serum levels of substance P (SP), vasoactive intestinal peptide (VIP), NO and NOS were detected by ELISA. The pathological changes of colon tissue were observed by HE staining. The c-kit level in colon tissue was detected by immunohistochemistry. The mRNA and protein levels of c-kit and Beclin1 in colon tissue were detected by PCR and Western blot.Results:Compared with the model group, the fecal moisture content, the carbon pushing distance and the intestinal pushing rate of the low, medium and high dose groups were significantly increased ( P<0.05), the serum SP level was increased ( P<0.05), the serum VIP, NO and NOS levels of the low, medium and high dose groups were decreased ( P<0.05), and the average expression score of c-kit in colon tissue of the low, medium and high dose groups was significantly increased ( P<0.05). The levels of c-kit mRNA (2.33 ± 0.35, 3.04 ± 0.17, 3.83 ± 0.23 vs. 0.61 ± 0.07) and protein (0.42 ± 0.06, 0.60 ± 0.07, 0.79 ± 0.08 vs. 0.22 ± 0.04)in colon tissue of rats in low, medium and high dose groups were increased ( P<0.05), and Beclin1 mRNA (4.17 ± 0.37, 3.35 ± 0.44, 1.05 ± 0.28 vs. 6.04 ± 0.31) and protein (0.76 ± 0.11, 0.57 ± 0.08, 0.43 ± 0.05 vs. 0.91 ± 0.06) were decreased ( P<0.05). Conclusion:Rhubarb dispensing granule could significantly increase the fecal moisture, intestinal motility rate, serum SP level and colonic tissue c-kit level, decrease serum VIP, NO, NOS level and colonic tissue Beclin1 level in rats with chronic transit constipation, and then inhibit autophagy of Cajal interstitial cells and regulate gastrointestinal motility in rats with chronic transit constipation.
ABSTRACT
Consistency in quality of traditional Chinese medicine granules is an important factor to ensure reproducible clinical efficacy. In this study rhubarb dispensing granules were utilized to construct an efficacious near-infrared spectroscopy (eNIRS) assay by combining NIRS and biopotency. A NIR method for assaying rhubarb dispensing particles was established, and information on different batches was collected. The diarrhea-inducing biopotency of rhubarb dispensing granules was determined based on a constipation model induced by diphenoxylate in mice. The animal protocol was approved by the Animal Ethic Committee of 302 Hospital of Chinese PLA People's Liberation Army (ID: IACUC-2019-0010). Ten anthraquinones were determined in rhubarb dispensing granules by UPLC. The correlation between NIR and biopotency was analyzed and five characteristic bands that correlated highly with bioactivity were identified, including 4 011-4 390, 4 859-5 461, 7 012-7 493, 10 992-11 312 and 11 871-12 489 cm-1. There were some differences in the main bands of different chemical constituents. In summary, five active bands based on NIRS were identified and found to be able to achieve rapid on-line detection of rhubarb dispensing granule quality. This research model may also provide reference for quality control of other Chinese medicine dispensing granules.
